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Introduction and purpose Diabetes is a disease resulting from impaired action or secretion of insulin. The number of patients currently amounts to approximately 422 million, and approximately 1.5 million deaths per year are directly attributed to this disease. Diabetes significantly reduces the quality of life and, if poorly controlled, can lead to serious complications. Mesenchymal stem cells are multipotent cells capable of differentiation. They have the ability to self-renew and have a modulating function. There are reports that they can be used in the treatment of this disease. The aim of the review was to present a new method of therapy and their possible effects. Material and methods The review was based on articles obtained from PubMed scientific database in the years 2015-2023, using the following keywords: diabetes mellitus, stem cells, diabetes complications. Results Implanted stem cells are able to transform into cells that produce and secrete insulin, and also enable better glycemic control. They can alleviate chronic inflammation and reduce fibrosis. Taking into account the complications that occur during long-term diabetes, the use of stem cells may be associated with improving the function of specific organs and tissues such as the kidneys, heart, eyes and nerves. Studies also report a positive effect of these cells on the healing process of wounds and ulcers. Conclusions Stem cells are a promising object of analysis. They can control glycemia and have a positive effect on the functioning of the kidneys, heart, eyes and nerves, and accelerate wound healing, but further, extensive research is needed to assess the effectiveness and safety of this therapy. [ABSTRACT FROM AUTHOR]

Background: Congenital bladder disorders in children necessitate innovative approaches for bladder tissue regeneration, aiming to minimize complications associated with conventional therapies. This study focused on generating a cell-seeded scaffold using superior smooth muscle cells (SMCs) by exploring the potential of smooth muscle cell spheroids (3D SMCs) compared to conventionally cultured SMCs (2D SMCs) for bladder tissue engineering. Additionally, adipose-derived stem cells (ADSCs) were investigated for their impact on SMC proliferation and maturation, and pre-differentiated smooth muscle-like ADSCs (pADSCs) for their potential as alternative cell source. Methods: 3D SMCs were seeded into a compressed collagen scaffold as monoculture and as co-culture with ADSCs or pADSCs and incubated for 2 weeks. Their contractile potential as well as proliferation and cell distribution within the scaffold were compared to conventionally cultured 2D SMCs by immunofluorescent staining and qRT-PCR. Results: 3D SMCs in collagen scaffolds exhibited significantly superior cell distribution, proliferation, and contractile marker expression compared to 2D SMCs. While ADSCs showed limited impact, co-culture with pADSCs enhanced contractile marker expression, though not surpassing 3D SMC monoculture. Conclusion: For the first time, a collagen scaffold seeded with 3D SMCs was generated and evaluated. This study recommends 3D SMCs as optimal building blocks for bladder tissue engineering, highlighting the potential of pADSCs as an alternative cell source. These findings offer crucial insights for refining cell sources as well as culture techniques in pediatric bladder regeneration and provide a superior cell-seeded scaffold for further bladder tissue engineering experiments. [ABSTRACT FROM AUTHOR]

Myocardial infarction, the most severe manifestation of coronary artery disease, is a leading cause of mortality worldwide. Here, a novel nanotherapy engineering approach is reported for enhancing therapeutic effects of mesenchymal stem cells (MSCs) on myocardial ischemia‐reperfusion (MI/R) injury. An anti‐oxidative and anti‐inflammatory nanotherapy (TPCD NP) is first fabricated. MSCs are successfully engineered with TPCD NP by endocytosis, and TPCD NP engineering does not affect stemness of MSCs. TPCD NP‐internalized MSCs (tn‐MSCs) are resistant to oxidative stress, cytotoxicity, and apoptosis induced by reactive oxygen species (ROS). Under oxidative distress, tn‐MSCs show stronger paracrine activity compared to MSCs. Consistently, tn‐MSCs effectively enhance angiogenesis of endothelial cells under pathological conditions. Also, tn‐MSCs protect cardiomyocytes from ROS‐induced cytotoxicity and apoptosis by improving mitochondrial membrane potential and regulating the p53 signaling pathway. In mice with MI/R injury, the survival time of tn‐MSCs in the injured heart is prolonged, compared to pristine MSCs. Correspondingly, tn‐MSCs more significantly reduce the infarct size, improve cardiac function, and promote cardiac remodeling in MI/R injury mice. Mechanistically, tn‐MSCs alleviate MI/R injury by attenuating oxidative stress and inflammation, inhibiting cardiomyocyte apoptosis, and promoting cardiac repair. Consequently, tn‐MSCs are promising for treating cardiovascular diseases linked to oxidative distress and inflammation. [ABSTRACT FROM AUTHOR]

ABSTRACT Background and Objective Methods and Results Conclusions CEFFE (Cell‐free fat extract) treatment for periocular fine lines requires thorough clinical evaluation to determine its efficacy and safety in enhancing skin quality. The research enrolled 10 healthy female participants aged 31–58, focusing on skin texture, elasticity, and barrier function.CEFFE treatment demonstrated significant benefits, with notable improvements observed as early as 3 months posttreatment, which continued throughout the 12‐month follow‐up period. Objective assessments revealed reductions in SEr% and SEw%, indicative of reduced skin roughness and wrinkles, particularly pronounced after the third month of treatment. Enhanced skin elasticity, as indicated by improvements in R2%, R5%, and R7%, was observed, with the most significant enhancements noted at the 6‐month follow‐up. Furthermore, TEWL decreased consistently, highlighting CEFFE's potential in maintaining the skin's barrier function and moisture retention. High patient satisfaction levels, with 70% expressing satisfaction ranging from satisfied to very satisfied, underscored CEFFE's clinical significance.CEFFE demonstrates potential as an effective and safe intervention for addressing periocular fine lines, providing a solution for fine lines while ensuring skin health (ChiCTR1900024329). [ABSTRACT FROM AUTHOR]

The beneficial effects of Neural Precursor Cell (NPC) transplantation in several neurological disorders are well established and they are generally mediated by the secretion of immunomodulatory and neurotrophic molecules. We therefore investigated whether Rett syndrome (RTT), that represents the first cause of severe intellectual disability in girls, might benefit from NPC-based therapy. Using in vitro co-cultures, we demonstrate that, by sensing the pathological context, NPC-secreted factors induce the recovery of morphological and synaptic defects typical of Mecp2 deficient neurons. In vivo, we prove that intracerebral transplantation of NPCs in RTT mice significantly ameliorates neurological functions. To uncover the molecular mechanisms underpinning the mediated benefic effects, we analyzed the transcriptional profile of the cerebellum of transplanted animals, disclosing the possible involvement of the Interferon γ (IFNγ) pathway. Accordingly, we report the capacity of IFNγ to rescue synaptic defects, as well as motor and cognitive alterations in Mecp2 deficient models, thereby suggesting this molecular pathway as a potential therapeutic target for RTT. Synopsis: To address the unmet need for a cure for RTT, the therapeutic potential of adult Neural Precursor Cells (NPCs) was investigated in vitro on Mecp2 deficient neurons and in vivo in transplanted Mecp2 deficient mice. NPC-secreted molecules rescue typical phenotypes of Mecp2 deficient neurons in culture. NPCs do not require Mecp2 to exert their beneficial effects. Intracerebral transplantation of NPCs prolongs the lifespan of Mecp2 null mice and restores their memory and motor functions. RNA-sequencing studies on transplanted brains have indicated the activation of the IFNγ pathway as a possible mechanism of action. IFNγ treatment of Mecp2 deficient animals and neurons confirms the therapeutic efficacy of the cytokine for RTT. To address the unmet need for a cure for RTT, the therapeutic potential of adult Neural Precursor Cells (NPCs) was investigated in vitro on Mecp2 deficient neurons and in vivo in transplanted Mecp2 deficient mice. [ABSTRACT FROM AUTHOR]

Depression is a common affective disorder characterized by persistent low mood, diminished interest or pleasure in normally enjoyable activities, disturbances in sleep patterns, and suicidal ideation. Conventional treatments often yield unsatisfactory results and are associated with several adverse effects. However, emerging literature has highlighted the potential of stem cell (SC) transplantation as a promising avenue for treating depression owing to its favorable anti-inflammatory and neurotrophic properties. This review summarizes the therapeutic effects and underlying mechanisms associated with SC transplantation in depression, offering a conceptual framework for the future application of SCs in the clinical treatment of depression. [ABSTRACT FROM AUTHOR]

Thalassemia and leukemia and related blood disorders are approved for blood stem cell transplants in India, for a stem cell transplant to be successful, the human leukocyte antigen (HLA) complex located on the arm of chromosome six must be a match between the cord blood donor and the recipient. In the quest to find an exact blood stem cell match for an individual, the HLA becomes the node at the center of community genetics where the HLA match is sought (not necessarily successful) in the extended family, the same caste, language, and ethnic (both national and the diaspora) groups. By considering thalassemia as a case study, how do we understand personalized stem cell therapies within biobanking infrastructures in Chennai? How do social categories get entwined with biological materials like cord blood? [ABSTRACT FROM AUTHOR]

Stem cells regenerate differentiated cells to maintain and repair tissues and organs. They also replenish themselves, i.e. self-renew, to support a lifetime of regenerative capacity. Here we study the renewal of skeletal muscle stem cell (MuSC) during regeneration. The transcriptional co-factors TAZ/YAP (via the TEAD transcription factors) regulate cell cycle and growth while the transcription factor YY1 regulates metabolic programs for MuSC activation. We show that MPP7 and AMOT join TAZ and YY1 to regulate a selected number of common genes that harbor TEAD and YY1 binding sites. Among these common genes, Carm1 can direct MuSC renewal. We demonstrate that the L27 domain of MPP7 enhances the interaction as well as the transcriptional activity of TAZ and YY1, while AMOT acts as an intermediate to bridge them together. Furthermore, MPP7, TAZ and YY1 co-occupy the promoters of Carm1 and other common downstream genes. Our results define a renewal program comprised of two progenitor transcriptional programs, in which selected key genes are regulated by protein-protein interactions, dependent on promoter context. Synopsis: Renewal divisions of muscle stem cells are regulated by the methyltransferase CARM1. This study shows that Carm1 is transcriptionally regulated by the coordinated action of MPP7, AMOT, YAP/TAZ, and YY1. Mpp7 cKO and Amot cKO mice display similar muscle stem cell renewal defects. Mpp7 cKO, Amot cKO, and Yap/Taz double cKO transcriptomes reveal a common set of down-regulated genes, including Carm1. MPP7, AMOT, YAP/TAZ, and YY1 form a protein complex to activate Carm1 transcription. The L27 domain of MPP7 facilitates complex formation and contributes to the enhanced transcriptional activity. Renewal divisions of muscle stem cells are regulated by the methyltransferase CARM1. This study shows that Carm1 is transcriptionally regulated by the coordinated action of MPP7, AMOT, YAP/TAZ, and YY1. [ABSTRACT FROM AUTHOR]

Background: In diagnosing chronic orthopedic implant infections culture of sonicate represents a supplement to tissue cultures. However, the extent to which biofilm forms on implant surfaces and the degree of dislodgement of bacteria by sonication remains unclear. In this in vivo study using a low bacterial inoculum, we aimed to determine whether a variable effect of sonication could be observed in a standardized in vivo model. Materials and Methods: Seven Wistar rats underwent surgery with intramuscular implantation of two bone xenograft implants, each containing two steel plates. The grafts were inoculated with approximately 500 colony forming units (CFU) of Staphylococcus epidermidis ATCC 35984. After 20 days the rats were sacrificed, and the steel plates were removed from the bone grafts. Epifluorescence microscopy and scanning electron microscopy (SEM) were used to visualize biofilm formation and dislodgement on the plate surfaces. In addition to cultures of sonicate, a quantitative S. epidermidis specific PCR was developed for enumeration of bacteria. Results: A chronic, low-grade implant infection was successfully established, with all animals remaining in good health. All infected bone graft implants yielded abundant growth of S. epidermidis, with a median of 3.25 (1.6–4.6) × 10⁷ CFU per/graft. We were unable to distinguish infected plates from negative controls using epifluorescence microscopy. On infected plates small colonies of staphylococci were identified by SEM. The number of bacteria detected in the sonicate was low with 500 (100–2400) CFU/plate and 475 (140–1821) copies/plate by qPCR. The difference in area covered by fluorescent material before and after sonication was 10.1 (5.7–12.3) %, p = 0.018. Conclusion: Despite the pronounced infection in the surrounding tissue, only few bacteria were detected on the surface of the steel implants. This is evident from the minimal findings by SEM before sonication, as well as the very low CFU counts and DNA copies in the sonicate. Sonication did not show variable effectiveness, indicating it is a valuable addition to, but not a replacement for biopsy cultures in cases of implant-associated infections with low-virulence microorganisms. [ABSTRACT FROM AUTHOR]

Epidermal stem cells control homeostasis and regeneration of skin and hair. In the hair follicle (HF) bulge of mammals, populations of slow-cycling stem cells regenerate the HF during cyclical rounds of anagen (growth), catagen (regression), and telogen (quiescence). Multipotent epidermal cells are also present in the HF above the bulge area, contributing to the formation and maintenance of sebaceous gland and upper and middle portions of the HF. Here, we report that the transcription factor KROX20 is enriched in an epidermal stem cell population located in the upper/middle HF. Expression analyses and lineage tracing using inducible Krox20-CreERT showed that Krox20-lineage cells migrate out of this HF region and contribute to the formation of the bulge in the HF, serving as ancestors of bulge stem cells. In vivo depletion of these cells arrests HF morphogenesis. This study identifies a marker for an epidermal stem cell population that is indispensable for hair homeostasis. [ABSTRACT FROM AUTHOR]

AbstractThe inherent adaptability of human mesenchymal stromal cells (hMSCs) to differentiate into neural lineages provides a valuable resource for investigating potential neurotoxicity in humans. By harnessing the ability of hMSCs to transform into astrocytes, we can evaluate the effects of various agents on these vital cells. Our protocol employs hMSCs sourced from umbilical cord tissue, ensuring a readily available supply of high-quality cells. The hMSC-to-neural workflow encompasses six essential steps: hMSC culture, followed by the generation of embryoid bodies (EBs) from these cells on specialized surfaces. Next, EBs and cells are expanded in a growth-promoting medium, directing them toward neural lineages. Subsequent differentiation into immature astrocytes is achieved through the use of specific factors. The process continues with the maturation of EBs/cells into astrocyte-like cells (hALCs) under optimized conditions, culminating in the final development of hALCs in a specialized medium. This methodology yields cells that display astrocyte morphology and express characteristic markers such as GFAP and S100β. The protocol is efficient, requiring roughly 6 weeks to generate hALCs from primary hMSCs without genetic manipulation. The application of hMSCs in evaluating cell damage triggered by neurotoxicants like MeHg and MGO underscores their potential as a valuable component within a more extensive battery of neurotoxicity tests. [ABSTRACT FROM AUTHOR]

Acute myeloid leukemia (AML) is a severe blood cancer with less than 50 % long-term survival. Despite advancements in treatment options, relapse is still the major obstacle. The main reason of this problem is ineffective targeting of leukemic stem cells (LSCs), which play an important role in tumor development and relapse. In our previous studies, we found that casticin, the major polyphenolic component of Vitex trifolia’s fruit, targets both leukemic cells and LSCs without affecting healthy tissues. Therefore, in this study, we aimed to investigate the effect of casticin-mediated cell death in relation to the LSCs-favored survival pathways at gene and protein expression levels using in vitro LSC-like and parental leukemic cell models.We validated the LSC character of KG1a and KG1 cells (84.55 % CD34+, CD38- and 93.55 % CD34+, CD38+, respectively) by flow cytometry. For the investigation of casticin’s mechanism of action, we employed real time-PCR, western blotting and bioinformatics analyses.Our results showed an increase in cleaved PARP/β-actin ratio but no change in LC3BI/II and SQSTM/β-actin ratios. Our gene expression, bioinformatics and immunoblotting analyses represented significant decrease in Shh, Gli and Wnt levels. We also elucidated a possible crosstalk between Hedgehog and other oncogenic cascades via the Gli, Notch, YAP, p38, Mcl-1, and Myc proteins in casticin mediated anti-leukemic effect.In conclusion, we found that casticin induces apoptosis in both LSC-like and parental leukemia cells mainly by suppressing Shh signaling, which is crucial for LSC survival and AML relapse. [ABSTRACT FROM AUTHOR]

Intestinal stem cells (ISCs) are highly vulnerable to damage, being in a constant state of proliferation. Reserve stem cells repair the intestinal epithelium following damage-induced ablation of ISCs. Here, we report that the epigenetic regulator plant homology domain (PHD) finger protein 16 (PHF16) restores homeostasis of the intestinal epithelium after initial damage-induced repair. In Phf16 −/Y mice, revival stem cells (revSCs) showed defects in exiting the regenerative state, and intestinal crypt regeneration failed even though revSCs were still induced in response to tissue damage, as observed by single-cell RNA sequencing (scRNA-seq). Analysis of Phf16 −/Y intestinal organoids by RNA sequencing (RNA-seq) and ATAC sequencing identified that PHF16 restores homeostasis of the intestinal epithelium by inducing retinoic acid receptor (RAR)/retinoic X receptor (RXR) target genes through HBO1-mediated histone H3K14 acetylation, while at the same time counteracting YAP/TAZ activity by ubiquitination of CDC73. Together, our findings demonstrate the importance of timely suppression of regenerative activity by PHF16 for the restoration of gut homeostasis after acute tissue injury. [Display omitted] • PHF16 is required for intestinal regeneration following irradiation • PHF16 is essential for timely exit from the regenerative phase • The PHF16/HBO1 complex promotes the differentiation of revival stem cells • PHF16 functions as an E3 ligase for CDC73 to suppress the expression of fetal genes Ahn and Kim et al. report that PHF16 controls the timely termination of intestinal injury response and the completion of regeneration. PHF16 promotes the expression of differentiation genes through its interaction with the HBO1 complex, while also acting as an E3 ligase for CDC73 to suppress the expression of fetal genes. [ABSTRACT FROM AUTHOR]

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Cell and Gene therapy are referred to as advanced therapies that represent overlapping fields of regenerative medicine. They have similar therapeutic goals such as to modify cellular identity, improve cell function, or fight a disease. These two therapeutic avenues, however, possess major differences. While cell therapy involves introduction of new cells, gene therapy entails introduction or modification of genes. Furthermore, the aim of cell therapy is often to replace, or repair damaged tissue, whereas gene therapy is used typically as a preventive approach. Diabetes mellitus severely affects the quality of life of afflicted individuals and has various side effects including cardiovascular, ophthalmic disorders, and neuropathy while putting enormous economic pressure on both the healthcare system and the patient. In recent years, great effort has been made to develop cutting-edge therapeutic interventions for diabetes treatment, among which cell and gene therapies stand out. This review aims to highlight various cell- and gene-based therapeutic approaches leading to the generation of new insulin-producing cells as a topmost "panacea" for treating diabetes, while deliberately avoiding a detailed molecular description of these approaches. By doing so, we aim to target readers who are new to the field and wish to get a broad helicopter overview of the historical and current trends of cell- and gene-based approaches in β-cell regeneration. [ABSTRACT FROM AUTHOR]

Chronic pathological conditions often induce persistent systemic inflammation, contributing to neuroinflammatory diseases like Multiple Sclerosis (MS). MS is known for its autoimmune-mediated damage to myelin, axonal injury, and neuronal loss which drive disability accumulation and disease progression, often manifesting as cognitive impairments. Understanding the involvement of neural stem cells (NSCs) and neural progenitor cells (NPCs) in the remediation of MS through adult neurogenesis (ANG) and gliogenesis—the generation of new neurons and glial cells, respectively is of great importance. Hence, these phenomena, respectively, termed ANG and gliogenesis, involve significant structural and functional changes in neural networks. Thus, the proper integration of these newly generated cells into existing circuits is not only key to understanding the CNS's development but also its remodeling in adulthood and recovery from diseases such as MS. Understanding how MS influences the fate of NSCs/NPCs and their possible neuroprotective role, provides insights into potential therapeutic interventions to alleviate the impact of MS on cognitive function and disease progression. This review explores MS, its pathogenesis, clinical manifestations, and its association with ANG and gliogenesis. It highlights the impact of altered NSCs and NPCs' fate during MS and delves into the potential benefits of its modifications. It also evaluates treatment regimens that influence the fate of NSCS/NPCs to counteract the pathology subsequently. [ABSTRACT FROM AUTHOR]

Background Current guidelines advocate a shared decision-making process approach to erectile dysfunction management, and while there is growing interest in regenerative therapies such as stem cell therapy, platelet-rich plasma injections, and low-intensity shockwave therapy for erectile dysfunction, comparative data on the efficacy of these modalities are limited. Aim This systematic review and network meta-analysis aims to compare stem cell therapy, platelet-rich plasma injections, and low-intensity shockwave therapy for managing erectile dysfunction and quantify their impact on the International Index of Erectile Function (IIEF). Methods In January 2024, a systematic search of online databases was performed to identify randomized clinical trials related to stem cell therapy, platelet-rich plasma injections, and low-intensity shockwave therapy in erectile dysfunction. Eligible articles reported outcomes using the IIEF score. Data were inputted into Review Manager 5.4 for pairwise meta-analysis. Data were then used to build a network in R Studio. These networks were used to model 200 000 Markov Chains via MonteCarlo sampling. The results are expressed as standardized mean difference (SMD) with 95% credible intervals (CrI). Meta-regression was used to adjust for PDE5is use. Outcomes Impact on the International Index of Erectile Function. Results A total of 16 studies involving 907 patients were analyzed. The standardized mean difference (SMD) vs control for stem cell therapy was 0.92 [95% CrI –0.49, 2.3]. For platelet-rich plasma, the SMD vs control was 0.83 [95% CrI 0.15, 1.5], and for low-intensity shockwave therapy, the SMD vs control was 0.84 [95% CrI 0.49, 1.2]. When stratifying low-intensity shockwave therapy by dose, the SMD vs control at 0.15 mJ/mm2 was 1.1 [95% CrI 0.36, 1.9], while at 0.09 mJ/mm2, it was 0.75 [95% CrI 0.26, 1.2]. Meta-regression adjusting for the administration of PDE5 inhibitors yielded non-significant results. Clinical implications The findings suggest that stem cells, platelet-rich plasma, and low intensity shockwave therapy, particularly at 0.15 and 0.09 mJ/mm2, may offer improvements in erectile function. Strengths and limitations The strength is the robust statistical methods. Limitations are in heterogeneity in control groups and follow-up durations among included studies. Conclusion Shockwave therapy and platelet-rich plasma demonstrated statistically significant improvements, though the clinical relevance and extent of their impact remain questionable. Further research is necessary to determine the efficacy of stem cell therapies for erectile function. [ABSTRACT FROM AUTHOR]

Background: One-step cell-based techniques of cartilage repair that lead to restoration of durable chondral tissue and long-term maintenance of joint function are cost-effective and ideal for routine use. Purposes: To examine the long-term clinical outcomes, after a mean follow-up duration of 14 years, of cartilage repair in the knee using a hyaluronic acid–based scaffold in association with bone marrow aspirate concentrate (HA-BMAC) and to evaluate the effect of age, lesion characteristics, and associated treatments on the outcome of this cartilage repair method. Study Design: Case series; Level of evidence, 4. Methods: Patients were followed prospectively for a mean duration of 14.0 years after undergoing treatment of knee full-thickness articular cartilage injury using HA-BMAC. Clinical evaluation consisted of the patient-reported scoring tools of the visual analog scale and the Knee injury and Osteoarthritis Outcome Score, which were completed preoperatively and at the time of final follow-up. Results: A total of 26 patients with a mean age of 48.3 years (17 male, 9 female) and median chondral lesion size of 6.6 cm2 (range, 1-27 cm2) were followed prospectively. There were 3 treatment failures, and 1 patient who underwent medial compartment unicompartmental arthroplasty 12 years after HA-BMAC treatment of patellar chondral injury. Of the 22 remaining patients, after a mean final follow-up duration of 14.0 years (range, 12-16 years), the median visual analog scale score of 0.6 was significantly decreased from the preoperative median score of 5.0 (P <.001). The median Knee injury and Osteoarthritis Outcome Score Pain (92), Symptoms (86), Activities of Daily Living (96), Sports (85), and Quality of Life (88) subscale values were all increased compared with the preoperative scores (P ≤.001). There was no correlation of clinical outcome score and body mass index. Conclusion: One-step cartilage repair of full-thickness chondral defects in the knee using an HA-BMAC led to successful long-term clinical outcomes and maintenance of joint junction after a mean follow-up duration of 14 years. Long-term clinical success in active, nonobese patients has been uniformly demonstrated across a wide range of patient ages and lesion types, including cases of multicompartment involvement, treatment of associated conditions, and large or bipolar chondral lesions. [ABSTRACT FROM AUTHOR]

Microglia identity is determined by intersecting developmental and environmental transcriptional networks and can be induced by forced expression of key lineage transcription factors (TFs) in human pluripotent stem cells. Forward programming has emerged as a powerful tool for efficient bulk generation of microglia. Key determinants of forward programming are the choice of TFs, their mode of gene delivery, and environmental cues. Forward programming, classical directed differentiation, and blood monocyte reprogramming are diverse and complementary tools for the study of human microglia in vitro. Standardisation of human microglia cultures is key for the interpretation of future downstream analysis. Microglia play vital roles in embryonic and post-natal development, homeostasis, and pathogen defence in the central nervous system. Human induced pluripotent stem cell (hiPSC)-based methods have emerged as an important source for the study of human microglia in vitro. Classical approaches to differentiate hiPSCs into microglia suffer from limitations including extended culture periods, consistency, and efficiency. More recently, forward programming has arisen as a promising alternative for the manufacture of bulk quantities of human microglia. This review provides a comprehensive assessment of published forward programming protocols that are based on forced expression of key lineage transcription factors (TFs). We focus on the choice of reprogramming factors, transgene delivery methods, and medium composition, which impact induction kinetics and the resulting microglia phenotype. [ABSTRACT FROM AUTHOR]

Due to the limitations of organ transplantation and the urgent need for treatment of chronic diseases, the benefit of stem cells for treatment has been studied and evaluated as an effective approach worldwide. One of the leading countries in this field is Iran. In this respect, several research and treatment institutes, including endocrinology and metabolism research institute are active in the use of stem cells in Iran. Herein, the aim is to review strategies, challenges, and opportunities for stem cell research and treatment in endocrinology and metabolism research institute. [ABSTRACT FROM AUTHOR]

Gastrulation is a key milestone in the development of an organism. It is a period of cell proliferation and coordinated cellular rearrangement, that creates an outline of the body plan. Our current understanding of mammalian gastrulation has been improved by embryo culture, but there are still many open questions that are difficult to address because of the intrauterine development of the embryos and the low number of specimens. In the case of humans, there are additional difficulties associated with technical and ethical challenges. Over the last few years, pluripotent stem cell models are being developed that have the potential to become useful tools to understand the mammalian gastrulation. Here we review these models with a special emphasis on gastruloids and provide a survey of the methods to produce them robustly, their uses, relationship to embryos, and their prospects as well as their limitations. [ABSTRACT FROM AUTHOR]

Parkinson's disease (PD) is characterized by the loss of the dopaminergic nigrostriatal pathway which has led to the successful development of drug therapies that replace or stimulate this network pharmacologically. Although these drugs work well in the early stages of the disease, over time they produce side effects along with less consistent clinical benefits to the person with Parkinson's (PwP). As such there has been much interest in repairing this pathway using transplants of dopamine neurons. This work which began 50 years ago this September is still ongoing and has now moved to first in human trials using human pluripotent stem cell‐derived dopaminergic neurons. The results of these trials are eagerly awaited although proof of principle data has already come from trials using human fetal midbrain dopamine cell transplants. This data has shown that developing dopamine cells when transplanted in the brain of a PwP can survive long term with clinical benefits lasting decades and with restoration of normal dopaminergic innervation in the grafted striatum. In this article, we discuss the history of this field and how this has now led us to the recent stem cell trials for PwP. [ABSTRACT FROM AUTHOR]

The discovery of mouse embryonic stem cells in 1981 transformed research in mammalian developmental biology and functional genomics. The subsequent generation of human pluripotent stem cells (PSCs) and the development of molecular reprogramming have opened unheralded avenues for drug discovery and cell replacement therapy. Here, I review the history of PSCs from the perspective that long‐term self‐renewal is a product of the in vitro signaling environment, rather than an intrinsic feature of embryos. I discuss the relationship between pluripotent states captured in vitro to stages of epiblast in the embryo and suggest key considerations for evaluation of PSCs. A remaining fundamental challenge is to determine whether naïve pluripotency can be propagated from the broad range of mammals by exploiting common principles in gene regulatory architecture. [ABSTRACT FROM AUTHOR]

Organoids are quickly becoming an accepted model for understanding human biology and disease. Pluripotent stem cells (PSC) provide a starting point for many organs and enable modeling of the embryonic development and maturation of such organs. The foundation of PSC‐derived organoids can be found in elegant developmental studies demonstrating the remarkable ability of immature cells to undergo histogenesis even when taken out of the embryo context. PSC‐organoids are an evolution of earlier methods such as embryoid bodies, taken to a new level with finer control and in some cases going beyond tissue histogenesis to organ‐like morphogenesis. But many of the discoveries that led to organoids were not necessarily planned, but rather the result of inquisitive minds with freedom to explore. Protecting such curiosity‐led research through flexible funding will be important going forward if we are to see further ground‐breaking discoveries. [ABSTRACT FROM AUTHOR]

Many strands of research by different groups, starting from teratocarcinomas in the laboratory mouse, later moving the corresponding human tumors, contributed to the isolation and description of human pluripotent stem cells (PSCs). In this review, I highlight the contributions from my own research, particularly at the Wistar Institute during the 1980s, when with my colleagues we characterized one of the first clonal lines of pluripotent human embryonal carcinoma (EC) cells, the stem cells of teratocarcinomas, and identified key features including cell surface antigen markers that have since found a place in the study and exploitation of human PSC. Much of this research depended upon close teamwork with colleagues, many in other laboratories, who contributed different expertise and experience. It was also often driven by circumstance and chance rather than pursuit of a grand design. [ABSTRACT FROM AUTHOR]

Stem cell research is the product of cumulative, integrated effort between and within laboratories and disciplines. The many collaborative steps that lead to that special "Eureka moment", when something that has been a puzzle perhaps for years suddenly become clear, is among the greatest pleasures of a scientific career. In this essay, the serendipitous pathway from first acquaintance with pluripotent stem cells to advanced cardiovascular models that emerged from studying development and disease will be described. Perhaps inspiration for later generations of stem cell researchers simply to follow whatever they find interesting. [ABSTRACT FROM AUTHOR]

The relationship of embryonal carcinoma (EC) cells, the stem cells of germ cell‐ or embryo‐derived teratocarcinoma tumors, to early embryonic cells came under intense scrutiny in the early 1970s when mouse chimeras were produced between EC cells and embryos. These chimeras raised tantalizing possibilities and high hopes for different areas of research. The normalization of EC cells by the embryo lent validity to their use as in vitro models for embryogenesis and indicated that they might reveal information about the relationship between malignancy and differentiation. Chimeras also showed the way for the potential introduction of genes, selected in EC cells in vitro, into the germ line of mice. Although EC cells provided material for the elucidation of early embryonic events and stimulated many studies of early molecular differentiation, after years of intense scrutiny, they fell short as the means of genetic manipulation of the germ line, although arguably they pointed the way to the development of embryonic stem (ES) cells that eventually fulfilled this goal. [ABSTRACT FROM AUTHOR]

• The Life cell derived from amniotic membrane promote endometrial cell growth. • The Life cell derived from amniotic membrane enhance the expression of genes associated with endometrial receptivity. • The LifeCell is a potential option for patients with treatment-resistant thin endometrium in cases of infertility. Successful assisted reproductive technology (ART) requires a receptive endometrium with appropriate thickness and the presence of specific cytokines, chemokines, and growth factors. Despite advancements in ART, the success rates remain suboptimal, particularly in individuals with thin endometrium resistant to treatment. In this study, we evaluated the potential effects of LifeCell, a product of BioNano Technology, on the growth, development, and acceptance of endometrial cells. We cultured endometrial cells in a defined medium with different concentrations of LifeCell and examined cell growth, development, and the expression of genes involved in endometrial receptivity. Co-culture of primary human endometrial cells with 5 % Life cell solution significantly stimulated the endometrial cell growth, development and receptivity genes expression. The expression levels of FGF2 and CSF in the 72 h co-cultured were significantly increased compared with other groups (P < 0.01). HOXA10 and LIF significantly increased in the 72 h co-cultured compared with 24 h co-cultured and control groups but had no significant level compared with 48 h cocultured. HOXA10 significantly increased in the 48 h cocultured compared with control group. IL-6 and Hb-EGF increased in the 48 h co-cultured compared to other groups but had no significant level. VEGF increased in the treated groups compared to control but had no significant level. The expression of OPN, unlike the other genes, decreased in the treated group compared to the control, which was not significant. These findings suggest that LifeCell may be a potential option for patients with treatment-resistant thin endometrium in cases of infertility. [ABSTRACT FROM AUTHOR]

Background: With the recent Food & Drug Administration (FDA) approval of cellular therapy that requires product manipulation prior to administration in combination with a short stability window, the need was identified for local dose preparation within the pharmacy rather than the off-site stem cell processing laboratory. This approval gave rise to assessment of regulatory standards surrounding cellular therapy, evaluation and revision of current standard operating procedures and policies with formal process validation, assessment of occupational exposure mitigation and safety considerations, and development of staff training and education. Objective: To describe and provide insight into the stepwise process of FACT validation and onboarding of commercially available cellular therapy products that require sterile compounding manipulation within a pharmacy prior to administration. Discussion: A multidisciplinary effort is required to attain FACT certification and implement pharmacist compounding of cellular therapy products. 1 Local preparation within a pharmacy facilitates a sound operational workflow and provides a pathway to perform aseptic manipulations of cellular therapy products safely and efficiently. Conclusion: Safe and successful administration of cellular therapies handled and compounded by pharmacy department staff along with program validation requires a preemptive review utilizing a multidisciplinary approach for process development. This manuscript will provide a foundation based on consistency and transparency in effective cellular therapy sterile compounding and aseptic manipulation, proper handling and disposal procedures, increased communication through creation and optimization of treatment plans and order-sets, standardized medical center staff education, and development of policies and standard operating procedures for the entire health care team. [ABSTRACT FROM AUTHOR]

Cardiac-expressed long noncoding RNAs (lncRNAs) are important for cardiomyocyte (CM) differentiation and function. Several lncRNAs have been identified and characterized for early CM lineage commitment, however those in later CM lineage specification and maturation remain less well studied. Moreover, unique atrial / ventricular lncRNA expression has never been studied in detail. Here, we characterized a novel ventricular myocyte-restricted lncRNA, not expressed in atrial myocytes, and conserved only in primates. First, we performed single cell RNA-seq on human pluripotent stem cell derived cardiomyocytes (hPSC-CM) at the late stages of 2, 6 and 12 weeks of differentiation. Weighted correlation network analysis identified core gene modules, including a set of lncRNAs highly abundant and predominantly expressed in the human heart. A lncRNA (we call VENTHEART , VHRT) co-expressed with cardiac maturation and ventricular-specific genes MYL2 and MYH7 , and was expressed in fetal and adult human ventricles, but not atria. CRISPR-mediated deletion of the VHRT gene led to impaired CM sarcomere formation and significant disruption of the ventricular CM gene program. Indeed, a similar disruption was not observed in VHRT KO hPSC-derived atrial CM, suggesting that VHRT exhibits only ventricular myocyte subtype-specific effects. Optical recordings validated that loss of VHRT significantly prolonged action potential duration at 90 % repolarization (APD 90) for ventricular-like, but not atrial-like, CMs. This reports the first lncRNA that is exclusively required for proper ventricular, and not atrial, CM specification and function. [Display omitted] • The VENTHEART lncRNA is co-expressed with cardiac genes during the process of cardiomyocyte development and maturation. • Deletion of the VENTHEART genomic locus leads to defects in ventricular cardiomyocyte function. • The VENTHEART lncRNA is characterized by chamber-specific expression and functionality in cardiac development. [ABSTRACT FROM AUTHOR]

Hypertranscription occurs when cells upregulate gene expression across the majority of the transcriptome, including at housekeeping and other ubiquitously expressed genes. Although masked by standard normalization procedures, hypertranscription can be detected using modifications to sample preparation and normalization of sequencing data. Originally characterized in embryonic stem cells, recent advances have identified hypertranscription to be pervasive in stem cells across development, reprogramming, organ homeostasis, and tissue regeneration. These contexts share unified mechanistic features of hypertranscription, many of which are redeployed in hypertranscribing cancer cells. As a counterpart to the hypertranscriptive state, emerging evidence suggests that global downregulation of transcriptional output to enter hypotranscription is a key component of embryonic diapause and stem cell quiescence, possibly also being co-opted by dormant cancer cells to survive chemotherapy. Stem cells are the fundamental drivers of growth during development and adult organ homeostasis. The properties that define stem cells – self-renewal and differentiation – are highly biosynthetically demanding. In order to fuel this demand, stem and progenitor cells engage in hypertranscription, a global amplification of the transcriptome. While standard normalization methods in transcriptomics typically mask hypertranscription, new approaches are beginning to reveal a remarkable range in global transcriptional output in stem and progenitor cells. We discuss technological advancements to probe global transcriptional shifts, review recent findings that contribute to defining hallmarks of stem cell hypertranscription, and propose future directions in this field. [ABSTRACT FROM AUTHOR]

Introduction: The complex nature of xerostomia prevents the establishment of a definite cure. Recently, research has pivoted towards stem cell transplantation for glandular reconstruction. The aim of this study is to provide an updated review of the existing research, to highlight the encountered challenges and research pathways, potentially enhancing the therapeutic applications of stem cell transplantation. Methods: This is a systematic review according to PRISMA guidelines, using the following databases: PubMed (PMC), PMC Europe, Scopus, Medline, Research Gate, Elsevier. The main question was whether stem cell therapies can contribute to the treatment of xerostomia. Results: 19 of 226 publications met the criteria for this review, including 'in vivo', 'in vitro' studies and clinical trials. All 19 studies described thoroughly the stem cell source and the transplantation method, and documented results based on analytical and statistical methods of confirmation. Data show that the various sources of stem cells play a significant role, with bone marrow or adipose tissue-derived pluripotent blasts being the most utilized. Human transplants in mice have also been accepted and reversed hyposalivation. The effects have been beneficial especially in models undergone radiotherapy (IR) or exhibit Sjogren Syndrome-like symptoms (SS), suggesting that with appropriate treatment and enrichment techniques, stem cell transplantation seems effective regardless of the cause of the disorder. Extracts and co-cultures of gland and stem cells also seem to improve gland function. Conclusion: Although in its initial stages, the use of stem cells seems to be a promising therapy to alleviate xerostomia regardless of its cause. [ABSTRACT FROM AUTHOR]

Objectives: Dental pulp stem cells (DPSCs) have been extensively used for tissue regeneration owing to their notable capabilities. Insulin‐like growth factor‐binding protein 5 (IGFBP5) regulates osteogenic differentiation of mesenchymal stem cells (MSCs); however, the underlying regulatory mechanisms require further investigation. Materials and Methods: Carboxyfluorescein succinimidyl ester, an alkaline phosphatase (ALP) activity assay and Alizarin Red staining were used to reveal the role of IGFBP5 in DPSCs. Protein expression levels were determined using western blotting. Immunofluorescence was used to observe cell sub‐localisation. Subcutaneous transplantation in nude mice was used to observe the osteogenesis of DPSCs in vivo. Results: IGFBP5 enhanced the proliferation and osteogenic differentiation of DPSCs. Deletion of the nuclear localisation sequence (NLS) of IGFBP5 prevented its nuclear import and abolished all its promoting effects on DPSCs; ivermectin stimulation attenuated the enhancement of ALP activity by IGBFP5. Bone‐like tissue formation promoted by IGFBP5 in vivo vanishes when the NLS is deleted. Inhibition of IGFBP5 nuclear import attenuated the IGFBP5‐induced phosphorylation of JNK (p‐JNK) and phosphorylated ERK (p‐ERK) in DPSCs. Conclusion: Our findings suggest that cell proliferation and osteogenic differentiation effects exerted by IGFBP5 on DPSCs are closely associated with their entry into the nucleus, thereby providing a novel potential target for tissue regeneration. [ABSTRACT FROM AUTHOR]

Pax‐6 emerges as a critical transcription factor that guides the fate of stem cells towards neural lineages. Its expression influences the differentiation of neural progenitors into diverse neuronal subtypes, glial cells, and other neural cell types. Pax‐6 operates with other regulatory factors to ensure the precise patterning and organization of the developing nervous system. The intricate interplay between Pax‐6 and other signaling pathways, transcription factors, and epigenetic modifiers underpins the complicated balance between stem cell maintenance, proliferation, and differentiation in neuroectodermal and ocular contexts. Dysfunction of Pax‐6 can lead to a spectrum of developmental anomalies, underscoring its importance in these processes. This review highlights the essential role of Pax‐6 expression in neuroectodermal and ocular stem cells, shedding light on its significance in orchestrating the intricate journey from stem cell fate determination to the emergence of diverse neural and ocular cell types. The comprehensive understanding of Pax‐6 function gained from a developmental biology perspective offers valuable insights into normal development and potential therapeutic avenues for neuroectodermal and ocular disorders. [ABSTRACT FROM AUTHOR]

Human allogeneic liver-derived progenitor cells (HALPCs) display advanced ability to differentiate into hepatocyte-like cells and exhibit potent immunomodulatory, anti-inflammatory, and anti-fibrotic properties. HALPCs have been successfully manufactured under good manufacturing practice (GMP) and are currently in clinical development. A previous phase 2a trial demonstrated the safety of peripheral intravenous infusions of HALPCs and preliminary evidence of the cells' properties to restore liver function in patients with acute-on-chronic liver failure (ACLF), thus potentially improving their survival. A phase 2b trial is currently ongoing across multiple centers (NCT04229901) to obtain proof-of-concept on efficacy and additional safety. HALPCs are currently manufactured using fetal bovine serum (FBS), which can reveal qualitative and quantitative variations between batches. The use of serum-free medium (SFM) represents an alternative means to overcome this variability while also complying fully with regulations. The aim of this study was to compare current FBS-containing culture conditions with two industry-available GMP-compliant SFMs: StemMACS (Miltenyi Biotec, Bergisch Gladbach, Germany) and PRIME-XV (FUJIFILM Irvine Scientific, Santa Ana, California, USA). The proliferation of HALPCs was significantly stimulated by both SFMs, which shortened both their emergence period and population doubling time. This effect was correlated with a significant improvement in their genetic stability as analyzed by conventional karyotyping. The expression profile (identity and purity) and functionality of HALPCs cultured in SFM were maintained, as demonstrated by flow cytometry and enzyme-linked immunoassay (ELISA), respectively. Their potency, evaluated via prostaglandin E2 (PGE2) secretion, showed a similar effect on CD4+ T-cell proliferation in FBS and SFM conditions. Furthermore, a greater proportion of HALPCs cultured in SFM showed enhanced expression of tissue factor (CD142) compared with the FBS condition. Altogether, SFM conditions enabled consistent HALPC quality to be achieved without altering their expression and functional profiles. [ABSTRACT FROM AUTHOR]

Aim: Angiogenesis is a key event in the successful healing of pulp injuries, and hypoxia is the main stimulator of pulpal angiogenesis. In this study, we investigated the effect of hypoxia on the proangiogenic potential of human dental pulp stem cells (hDPSCs) and the role of miR‐143‐5p in the process. Methodology: Human dental pulp stem cells were isolated, cultured and characterized in vitro. Cobalt chloride (CoCl2) was used to induce hypoxia in hDPSCs. CCK‐8 and Transwell assays were used to determine the effect of hypoxia on hDPSCs proliferation and migration. Quantitative real‐time polymerase chain reaction (qRT‐PCR), Western blotting (WB) and ELISA were performed to assess the mRNA and protein levels of HIF‐1α and angiogenic cytokines in hDPSCs. The effect of hypoxia on hDPSCs proangiogenic potential was measured in vitro using Matrigel tube formation and chick chorioallantoic membrane (CAM) assays. Recombinant lentiviral vectors were constructed to stably overexpress or inhibit miR‐143‐5p in hDPSCs, and the proangiogenic effects were assessed using qRT‐PCR, WB, and tube formation assays. miR‐143‐5p target genes were identified and verified using bioinformatics prediction tools, dual‐luciferase reporter assays and RNA pull‐down experiments. Finally, a subcutaneous transplantation model in nude mice was used to determine the effects of hypoxia treatment and miR‐143‐5p overexpression/inhibition in hDPSCs in dental pulp regeneration. Results: Hypoxia promotes hDPSCs proliferation, migration and proangiogenic potential. The in vivo experiments showed that hypoxia treatment (50 and 100 μM CoCl2) promoted pulp angiogenesis and dentine formation. In contrast to the levels of proangiogenic factors, miR‐143‐5p levels decreased with increasing CoCl2 concentration. miR‐143‐5p inhibition significantly promoted proangiogenic potential of hDPSCs, whereas miR‐143‐5p overexpression inhibited angiogenesis in vitro. Dual‐luciferase reporter assay identified retinoic acid receptor‐related orphan receptor alpha (RORA) as an miR‐143‐5p target gene in hDPSCs. RNA pull‐down experiments demonstrated that HIF‐1α and RORA were pulled down by biotin‐labelled miR‐143‐5p, and the levels of HIF‐1α and RORA bound to miR‐143‐5p in the hypoxia group were lower than those in the normoxia group. Inhibition of miR‐143‐5p expression in hDPSCs promoted ectopic dental pulp tissue regeneration. Conclusions: CoCl2‐induced hypoxia promotes hDPSCs‐driven paracrine angiogenesis and pulp regeneration. The inhibition of miR‐143‐5p upregulates the proangiogenic potential of hDPSCs under hypoxic conditions by directly targeting HIF‐1α and RORA. [ABSTRACT FROM AUTHOR]

Key message: The SBP-box transcription factor PlSPL2 silencing in herbaceous peony enhanced stem strength by regulating xylem development, whereas its overexpression in tobacco resulted in weaker stem strength and undeveloped xylem. The strength of plant stems is a critical determinant of lodging resistance of plants, which significantly affects crop yield and cut-flower quality. Squamosa promoter binding (SBP) protein-like (SPL) transcription factors (TFs), participate in multiple regulatory processes, particularly in stem development. In this study, PlSPL2, an orthologous gene of Arabidopsis AtSPL2 in herbaceous peony, was isolated and found to contain a conserved SBP domain featuring two typical Zn-binding sites, as well as a nuclear localization sequence (NLS). Subsequently, transient infection of tobacco leaf epidermal cells using Agrobacterium confirmed the nuclear localization of PISPL2 protein. Additionally, gene expression analyses revealed that PlSPL2 was preferentially expressed in stems, and demonstrated a download trend in expression levels within vascular bundles during stem cell wall development. Furthermore, silencing of PlSPL2 in herbaceous peony enhanced stem strength. The silenced plants exhibited more developed xylems with wider radii and higher numbers of cell layers. Overexpression of PlSPL2 in tobacco, however, resulted in weaker stem strength, accompanied by a narrower radius of the xylem. These findings suggested that PlSPL2 was a negative regulator of herbaceous peony stem development, and its discovery and research could significantly contribute to a deeper understanding of stem growth and development mechanisms. [ABSTRACT FROM AUTHOR]

Summary: Single‐cell proteomics (SCP) is an emerging approach to resolve cellular heterogeneity within complex tissues of multi‐cellular organisms.Here, we demonstrate the feasibility of SCP on plant samples using the model plant Arabidopsis thaliana. Specifically, we focused on examining isolated single cells from the cortex and endodermis, which are two adjacent root cell types derived from a common stem cell lineage.From 756 root cells, we identified 3763 proteins and 1118 proteins/cell. Ultimately, we focus on 3217 proteins quantified following stringent filtering. Of these, we identified 596 proteins whose expression is enriched in either the cortex or endodermis and are able to differentiate these closely related plant cell types.Collectivity, this study demonstrates that SCP can resolve neighboring cell types with distinct functions, thereby facilitating the identification of biomarkers and candidate proteins to enable functional genomics. See also the Commentary on this article by van Schie & Weijers, 244: 1678–1680. [ABSTRACT FROM AUTHOR]

PIEZO1 is a eukaryotic membrane protein that assembles as trimers to form calcium‐permeable, non‐selective cation channels with exquisite capabilities for mechanical force sensing and transduction of force into effect in diverse cell types that include blood cells, endothelial cells, epithelial cells, fibroblasts and stem cells and diverse systems that include bone, lymphatics and muscle. The channel has wide‐ranging roles and is considered as a target for novel therapeutics in ailments spanning cancers and cardiovascular, dental, gastrointestinal, hepatobiliary, infectious, musculoskeletal, nervous system, ocular, pregnancy, renal, respiratory and urological disorders. The identification of PIEZO1 modulators is in its infancy but useful experimental tools emerged for activating, and to a lesser extent inhibiting, the channels. Elementary structure–activity relationships are known for the Yoda series of small molecule agonists, which show the potential for diverse physicochemical and pharmacological properties. Intriguing effects of Yoda1 include the stimulated removal of excess cerebrospinal fluid. Despite PIEZO1's broad expression, opportunities are suggested for selective positive or negative modulation without intolerable adverse effects. Here we provide a focused, non‐systematic, narrative review of progress with this pharmacology and discuss potential future directions for research in the area. [ABSTRACT FROM AUTHOR]

Objective: The objective of this study was to identify the N6-methyladenosine (m6A)- circHECA molecule in secondary hair follicles (SHFs) of cashmere goats, and generate its potential regulatory network, as well as explore the potential relationship between transcriptional pattern of m6A-circHECA and promoter methylation of its host gene (HECA). Methods: The validation of circHECA m6A sites was performed using methylation immunoprecipitation (Me-RIP) along with reverse transcription-quantitative polymerase chain reaction (RT-qPCR) technique. The nucleus and cytoplasm localizations of m6A- circHECA were performed using SHF stem cells of cashmere goats with RT-qPCR analysis. Based on in-silico analysis, the regulatory networks of m6A-circHECA were generated with related signal pathway enrichment. The methylation level of promoter region of m6A-circHECA host gene (HECA) was assessed by the bisulfite sequencing PCR (BSPPCR) technique. Results: The m6A-circHECA was confirmed to contain four m6A modification sites including m6A-213, m6A-297, m6 A-780, and m6A-927, and it was detected mainly in cytoplasm of the SHF stem cells of cashmere goats. The integrated regulatory network analysis showed directly or indirectly complex regulatory relationships between m6A-circHECA of cashmere goats and its potential target molecules: miRNAs, mRNAs, and proteins. The regulatory network and pathway enrichment indicated that m6A-circHECA might play multiple roles in the SHF physiology process of cashmere goats through directly or indirectly interacting or regulating its potential target molecules. A higher methylation level of promoter region of HECA gene in SHFs of cashmere goats might cause the lower expression of m6A-circHECA. Conclusion: The m6A-circHECA might play multiple roles in SHF physiology process of cashmere goats through miRNA mediated pathways along with directly or indirectly interaction with its target proteins. The promoter methylation of m6A-circHECA host gene (HECA) most likely was implicated in its expression inhibition in SHFs of cashmere goats. [ABSTRACT FROM AUTHOR]

Introduction: Intercostal nerve transfer is a surgical technique used to restore function in patients with total brachial plexus injury. Stem cell and secretome therapy has been explored as a potential treatment for brachial plexus injuries. This study aimed to compare the functional and histologic outcome of intercostal nerve transfer to median nerve with local stem cells or secretome injection in total type brachial plexus injuries. Materials and methods: This was a double-blinded, randomized controlled study (RCT). We included patients with neglected total type brachial plexus injury (BPI) who underwent nerve transfer and local injection of either umbilical cord-derived mesenchymal stem cells (UC-MSC) or secretome into median nerve–flexor digitorum superficialis (FDS) neuromuscular junction (NMJ). We measured preoperative and 8-month postoperative FDS muscle strength, SF-36, DASH score, and histologic assessment. We then analyzed the difference outcome between those two groups. Result: A total of 15 patients were included in this study. Our study found that after nerve transfer and implantation with either UC-MSC or secretome, significant postoperative improvements were observed in physical functioning, role limitations, energy/fatigue, emotional well-being, social functioning, pain, general health, and DASH scores, particularly in the overall cohort and the secretome group. When we compared the mean difference of clinical outcome from preoperative to postoperative between UC-MSC and secretome groups, the UC-MSC group showed better improvement of health change in SF-36 subgroup compared to secretome group. From the analysis, there was no significant difference in the histologic outcomes (inflammation, regeneration, and fibrosis) in overall cohort between preoperative and postoperative cohort. There was also no significant difference in mean change of the histologic outcomes (inflammation, regeneration, and fibrosis) preoperative and postoperatively between UC-MSC and secretome groups. Discussion and conclusion: Implantation of either UC-MSC or secretome along with nerve transfer may provide clinical improvement, while to achieve histologic improvement, further conditioning should be performed. [ABSTRACT FROM AUTHOR]

Purpose: Periprosthetic joint infection is a complication of total joint arthroplasty with treatment costs over $1.6 billion dollars per year in the US with high failure rates. Therefore, generation of coatings that can prevent infection is paramount. Diamond-like carbon (DLC) is an ideal coating for implants as they are wear-resistant, corrosion-resistant, inert, and have a low friction coefficient. The purpose of this study was to test the efficacy of DLC surface treatment in prevention of biofilm on titanium discs infected with Staphylococcus aureus in vitro. Methods: Titanium alloy discs (n = 4 non-coated and n = 4 DLC-coated) were infected with 5 × 105 colony-forming units (CFU) of S. aureus for 2 weeks then analysed via crystal violet and scanning electron microscopy (SEM). Results: Crystal violet analysis yielded differences in the appearance of biofilm on implant surface where DLC-coated had a clumpier appearance but no difference in biofilm quantification. Interestingly, this clumpy appearance did lead to differences in SEM biofilm coverage where significantly less biofilm coverage was found on DLC-coated discs (81.78% vs. 54.17%, p < 0.003). Conclusion: DLC-coated titanium alloy implants may have preventative properties in S. aureus infection. Observing differences in biofilm coverage does warrant additional testing including CFU titration and biofilm kinetics with eventual use in an animal model of periprosthetic joint infection. [ABSTRACT FROM AUTHOR]

Acoustic manipulation or perturbation of biological soft matter has emerged as a promising clinical treatment for a number of applications within regenerative medicine, ranging from bone fracture repair to neuromodulation. The potential of ultrasound (US) endures in imparting mechanical stimuli that are able to trigger a cascade of molecular signals within unscathed cells. Particularly, low-intensity pulsed ultrasound (LIPUS) has been associated with bio-effects such as activation of specific cellular pathways and alteration of cell morphology and gene expression, the extent of which can be modulated by fine tuning of LIPUS parameters including intensity, frequency and exposure time. Although the molecular mechanisms underlying LIPUS are not yet fully elucidated, a number of studies clearly define the modulation of specific ultrasonic parameters as a means to guide the differentiation of a specific set of stem cells towards adult and fully differentiated cell types. Herein, we outline the applications of LIPUS in regenerative medicine and the in vivo and in vitro studies that have confirmed the unbounded clinical potential of this platform. We highlight the latest developments aimed at investigating the physical and biological mechanisms of action of LIPUS, outlining the most recent efforts in using this technology to aid tissue engineering strategies for repairing tissue or modelling specific diseases. Ultimately, we detail tissue-specific applications harnessing LIPUS stimuli, offering insights over the engineering of new constructs and therapeutic modalities. Overall, we aim to lay the foundation for a deeper understanding of the mechanisms governing LIPUS-based therapy, to inform the development of safer and more effective tissue regeneration strategies in the field of regenerative medicine. [ABSTRACT FROM AUTHOR]

Apoptosis is a regulated cell death that depends on caspases. It has mainly been studied as a mechanism for the removal of unwanted cells. However, apoptotic cells can induce fate or behavior changes of their neighbors and thereby participate in development. Here, we address the functions of apoptosis during metamorphosis of the cnidarian Hydractinia symbiolongicarpus. We describe the apoptotic profile during metamorphosis of the larva and identify Caspase3/7a , but no other executioner caspases, as essential for apoptosis in this context. Using pharmacological and genetic approaches, we find that apoptosis is required for normal head development. Inhibition of apoptosis resulted in defects in head morphogenesis. Neurogenesis was compromised in the body column of apoptosis-inhibited animals but there was no effect on the survival or proliferation of stem cells, suggesting that apoptosis is required for cellular commitment rather than for the maintenance of their progenitors. Differential transcriptomic analysis identifies TRAF genes as downregulated in apoptosis-inhibited larvae and functional experiments provide evidence that they are essential for head development. Finally, we find no major role for apoptosis in head regeneration in this animal, in contrast to the significance of apoptosis in Hydra head regeneration. [Display omitted] • A wave of apoptosis accompanies the initial phase of Hydractinia metamorphosis. • Caspase3/7a, but not other caspases, mediates apoptosis during metamorphosis. • Inhibition of apoptosis results in defective head development and neurogenesis. • Knockdown of TRAF genes phenocopies apoptosis inhibition. [ABSTRACT FROM AUTHOR]

The dentition is critical to animal survival and teeth are present in modern vertebrates including teleost fish, sharks, amphibians, mammals and reptiles. The developmental processes that give rise to teeth are not just preserved through evolution but also share high level of similarity with the embryogenesis of other ectodermal organs. In this review we go beyond the embryonic phase of tooth development to life-long tooth replacement. We will address the origins of successional teeth, the location of putative tissue-resident stem cells, how de novo tooth formation continues throughout life and how teeth are shed in a spatially and temporally controlled manner. We review the evidence that the dental epithelium, which is the earliest recognizable dental structure in the reptilian dentition, serves as a putative niche for tissue-resident epithelial stem cells and recent molecular findings from transcriptomics carried out in reptilian dentitions. We discuss how odontoclasts resorb the primary tooth allowing eruption of the successional tooth. The reptiles, particularly lizards, are emerging as some of the most accessible animals to study tooth replacement which has relevance to evolution of the dentition and human dental disorders. [Display omitted] • A review of the common origins of teeth and other ectodermal specializations. • Introduction to adult reptile teeth which undergo life-long organ regeneration. • Review of RNA expression profiling of reptilian dentitions and insights into the molecular mechanisms of tooth replacement. • A review of pulse-chase experiments to define the locations of tissue-resident, dental epithelial stem cells. • How comparative studies on reptile teeth contribute to the fields of evolution, organ replacement, and dental pathology. [ABSTRACT FROM AUTHOR]

Statement of the Problem: Success of pulpotomy of primary teeth depends on biological and cytotoxic effects of pulp capping agents. Mineral trioxide aggregate (MTA), Biodentine, calcium enriched mixture (CEM) cement, and ferric sulfate (FS) are among the commonly used pulp capping agents (PCAs) for pulpotomy, and their successful application has been previously evaluated. Purpose: This study aimed to compare the cytotoxicity of PCAs against mesenchymal stem cells isolated from human exfoliated deciduous teeth (SHEDs). Materials and Method: In this in vitro study, SHEDs were exposed to MTA, Biodentine, CEM cement, and FS for 24 and 72 hours. The methyl thiazolyl tetrazolium (MTT) assay was performed for five different concentrations of PCAs after 24 and 72 hours of exposure. Data were analyzed by ANOVA. Results: Generally, the biocompatibility increased by reduction in concentration. All tested concentrations showed higher biocompatibility at 72 hours compared with 24 hours (p< 0.0001). Comparison of cytotoxicity of different biomaterials revealed no significant difference at any time point (p> 0.05). Conclusion: In general, the cytotoxicity of MTA, Biodentine, CEM cement, and FS was comparable, with no significant difference. Cytotoxicity decreased over time and by a reduction in concentration of biomaterials. MTA and Biodentine showed maximum biocompatibility followed by FS, and CEM cement. [ABSTRACT FROM AUTHOR]

Traditional therapeutic approaches in the treatment of cancer have many side effects and are often ineffective and non-specific, leading to the development of therapy-resistant tumour cells. Recently, numerous discoveries about stem cells have given a new outlook on their application in oncology. Stem cells are unique because of their biological attributes, including self-renewal, differentiation in different types of specialized cells and synthesis of molecules that interplay with tumour niche. They are already used as an effective therapeutic option for haematological malignancies, such as multiple myeloma and leukaemia. The main goal of this study is to investigate the possible applications of different types of stem cells in cancer treatment and to summarize novel advances, as well as the limitations of their application in cancer treatment. Research and clinical trials that are underway revealed and confirmed the enormous potential of regenerative medicine in the treatment of cancer, especially when combined with different nanomaterials. Nanoengineering of stem cells has been the focus of novel studies in the area of regenerative medicine, such as the production of nanoshells and nanocarriers that enhance the transport and uptake of stem cells in their targeted tumour niche and enable the effective monitoring of stem cell effects on tumour cells. Although nanotechnology has a lot of limitations, it provides new opportunities for the development of effective and innovative stem cell therapies. [ABSTRACT FROM AUTHOR]

Aims: Allogeneic stem cell therapy is more logistically suitable compared with autologous cell therapy for large‐scale patient treatment. We aim to investigate the clinical safety and efficacy profile of the allogeneic adipose tissue derived mesenchymal stromal cell product (CSCC_ASC) as an add‐on therapy in patients with chronic non‐ischaemic heart failure with reduced left ventricular ejection fraction (HFrEF) < 40%. Methods and results: This is a single‐centre investigator‐initiated randomized phase I/II study with direct intra‐myocardial injections of 100 million allogeneic CSCC_ASC. A total of 30 HFrEF patients with New York Heart Association (NYHA) class ≥II despite optimal anticongestive heart failure medication and plasma NT‐proBNP > 300 pg/mL (>35 pmol/L) were included and randomized 2:1 to CSCC_ASC or standard care. The primary endpoint left ventricular end systolic volume (LVESV) and other echo related parameters were analysed by an investigator blinded for treatment allocation. No difference in serious adverse events was observed between groups. LVESV decreased significantly from baseline to 6 months follow‐up in the ASC group (153.7 ± 53.2 mL and 128.7 ± 45.6 mL, P < 0.001) and remained unchanged in the standard care group (180.4 ± 39.4 mL and 186.7 ± 48.9 mL, P = 0.652). There was a significant difference between the groups in LVESV change (31.3 ± 11.0 mL, P = 0.009). The difference from baseline to follow‐up between the two groups in left ventricular end diastolic volume (LVEDV) was 18.7 ± 12.4 mL, P = 0.146 and in left ventricular ejection fraction (LVEF) −7.8 ± 2.1%, P = 0.001. Considering the baseline values of LVESV, LVEDV and LVEF as covariates, the difference between groups for change from baseline to follow‐up resulted in a P‐value of 0.056, 0.076, and 0.738, respectively. NYHA class and self‐reported health did also improve significantly in the ASC group compared with the standard care group (0.7 ± 0.2, P = 0.001 and −12.8 ± 5.3, P = 0.025; respectively). There was no difference in NT‐proBNP (−371 ± 455 pmol/L, P = 0.422) or in 6 min walk test (12 ± 31 m, P = 0.695) between groups. Conclusions: Intramyocardial injections of allogeneic CSCC_ASC in patients with chronic non‐ischaemic HFrEF was safe and improved LVESV, LVEF, NYHA class, and self‐reported health compared with standard care group. [ABSTRACT FROM AUTHOR]

Regeneration of orofacial tissues is hampered by the lack of adequate vascular supply. Implantation of in vitro engineered, prevascularized constructs has emerged as a strategy to allow the rapid vascularization of the entire graft. Given the angiogenic properties of dental pulp stem cells, we hereby established a preclinical model of prevascularized constructs loaded with stem cells from human exfoliating deciduous teeth (SHED) in a 3-dimensional-printed material and provided a functional analysis of their in vivo angiogenesis, vascular perfusion, and permeability. Three different cell-loaded collagen hydrogels (SHED–human umbilical vein endothelial cell [HUVEC], HUVEC with SHED-conditioned medium, and SHED alone) were cast in polylactic acid (PLA) grids and ectopically implanted in athymic mice. At day 10, in vivo positron emission tomography (PETscan) revealed a significantly increased uptake of radiotracer targeting activated endothelial cells in the SHED-HUVEC group compared to the other groups. At day 30, ex vivo micro–computed tomography imaging confirmed that SHED-HUVEC constructs had a significantly increased vascular volume compared to the other ones. Injection of species-specific lectins analyzed by 2-photon microscopy demonstrated blood perfusion of the engineered human vessels in both prevascularized groups. However, in vivo quantification showed increased vessel density in the SHED-HUVEC group. In addition, coinjection of fluorescent lectin and dextran revealed that prevascularization with SHED prevented vascular leakage, demonstrating the active role of SHED in the maturation of human-engineered microvascular networks. This preclinical study introduces a novel PLA prevascularized and implantable construct, along with an array of imaging techniques, to validate the ability of SHED to promote functional human-engineered vessels, further highlighting the interest of SHED for orofacial tissue engineering. Furthermore, this study validates the use of PETscan for the early detection of in vivo angiogenesis, which may be applied in the clinic to monitor the performance of prevascularized grafts. [ABSTRACT FROM AUTHOR]

Approximately 5%–10% of fractures go on to delayed healing and nonunion, posing significant clinical, economic, and social challenges. Current treatment methods involving open bone harvesting and grafting are associated with considerable pain and potential morbidity at the donor site. Hence, there is growing interest in minimally invasive approaches such as bone marrow aspirate concentrate (BMAC), which contains mesenchymal stromal cells (MSCs), macrophages (Mφ), and T cells. However, the use of cultured or activated cells for treatment is not yet FDA‐approved in the United States, necessitating further exploration of optimal cell types and proportions for effective bone formation. As our understanding of osteoimmunology advances, it has become apparent that factors from anti‐inflammatory Mφ (M2) promote bone formation by MSCs. Additionally, M2 Mφ promote T helper 2 (Th2) cells and Treg cells, both of which enhance bone formation. In this study, we investigated the interactions among MSCs, Mφ, and T cells in bone formation and explored the potential of subsets of BMAC. Coculture experiments were conducted using primary MSCs, Mφ, and CD4+ T cells at specific ratios. Our results indicate that nonactivated T cells had no direct influence on osteogenesis by MSCs, while coculturing MSCs with Mφ and T cells at a ratio of 1:5:10 positively impacted bone formation. Furthermore, higher numbers of T cells led to increased M2 polarization and a higher proportion of Th2 cells in the early stages of coculture. These findings suggest the potential for enhancing bone formation by adjusting immune and mesenchymal cell ratios in BMAC. By understanding the interactions and effects of immune cells on bone formation, we can develop more effective strategies and protocols for treating bone defects and nonunions. Further studies are needed to investigate these interactions in vivo and explore additional factors influencing MSC‐based therapies. [ABSTRACT FROM AUTHOR]