Search

Juvenile osteochondritis dissecans (JOCD) is a pediatric disease, which begins with an osteonecrotic lesion in the secondary ossification center which, over time, results in the separation of the necrotic fragment from the parent bone. JOCD predisposes to early-onset osteoarthritis. However, the knowledge gap in JOCD pathomechanisms severely limits current therapeutic strategies. To elucidate its etiology, we conducted a study with induced pluripotent stem cells (iPSCs) from JOCD and control patients. iPSCs from skin biopsies were differentiated to iMSCs (iPSC-derived mesenchymal stromal cells) and subjected to chondrogenic and endochondral ossification, and endoplasmic reticulum (ER)-stress induction assays. Our study, using 3 JOCD donors, showed that JOCD cells have lower chondrogenic capability and their endochondral ossification process differs from control cells; yet, JOCD- and control-cells accomplish osteogenesis of similar quality. Our findings show that endoplasmic reticulum stress sensing and response mechanisms in JOCD cells, which partially regulate chondrocyte and osteoblast differentiation, are related to these differences. We suggest that JOCD cells are more sensitive to ER stress than control cells, and in pathological microenvironments, such as microtrauma and micro-ischemia, JOCD pathogenesis pathways may be initiated. This study is the first, to the best of our knowledge, to realize the important role that resident cells and their differentiating counterparts play in JOCD and to put forth a novel etiological hypothesis that seeks to consolidate and explain previously postulated hypotheses. Furthermore, our results establish well-characterized JOCD-specific iPSC-derived in vitro models and identified potential targets which could be used to improve diagnostic tools and therapeutic strategies in JOCD.

After more than a century of political and economic integration, Southern Ireland exited the United Kingdom in 1922. By identifying the leading business firms of the era and the political and religious allegiances of their owners, this paper explores the perspective of the Southern Irish business establishment on the issues involved. While the mass of the population was Catholic and by 1918 favored secession, the business elite is shown to have been predominantly Protestant and strongly supportive of continued integration. Business elite perceptions of the consequences of exiting the United Kingdom are explored, and post-independence economic and business developments assessed in light of the concerns expressed at the time. The paper also charts the post-independence fate of the leading former unionist firms and the erosion and eventual disappearance of the sectarian divisions then prevalent in Irish business life.

Belief in the benefits of industrial protection had long been a cornerstone of nationalist ideology. Cumann na nGaedheal followed a policy of selective protection while Fianna Fáil was ideologically committed not just to import-substituting industrialisation but to as high a degree of self-sufficiency as possible. The Departments of Finance and Industry and Commerce differed sharply on the costs and benefits of trade restrictions. This article explores the perspective of the Department of Finance and in particular that of J. J. McElligott, Assistant Secretary from 1923 and Secretary of the Department from 1927 to 1953. It demonstrates the strong continuity between his position and that of T. K. Whitaker, who became Secretary in 1956 and whose 1958 report on Economic Development is widely credited with providing the intellectual foundation for the trade liberalisation process of the following decades.

There has been considerable interest in the generation of functional mesenchymal stromal cell (MSC) preparations from induced pluripotent stem cells (iPSCs) and this is now regarded as a potential source of unlimited, standardized, high-quality cells for therapeutic applications in regenerative medicine. Although iMSCs meet minimal criteria for defining MSCs in terms of marker expression, there are substantial differences in terms of trilineage potential, specifically a marked reduction in chondrogenic and adipogenic propensity in iMSCs compared with bone marrow-derived (BM) MSCs. To reveal the cellular basis underlying these differences, we conducted phenotypic, functional, and genetic comparisons between iMSCs and BM-MSCs. We found that iMSCs express very high levels of both KDR and MSX2 compared with BM-MSCs. In addition, BM-MSCs had significantly higher levels of PDGFRα. These distinct gene expression profiles were maintained during culture expansion, suggesting that prepared iMSCs are more closely related to vascular progenitor cells (VPCs). Although VPCs can differentiate along the chondrogenic, osteogenic, and adipogenic pathways, they require different inductive conditions compared with BM-MSCs. These observations suggest to us that iMSCs, based on current widely used preparation protocols, do not represent a true alternative to primary MSCs isolated from BM. Furthermore, this study highlights the fact that high levels of expression of typical MSC markers such as CD73, CD90, and CD105 are insufficient to distinguish MSCs from other mesodermal progenitors in differentiated induced pluripotent stem cell cultures. Stem Cells 2019;37:754–765

Traumatic spinal cord injury (SCI) results in complex neurological dysfunction. Astrocytes react in response to SCI by undergoing proliferation and migrating to the injury site, resulting in astrogliosis. Neural stem cells (NSCs) have been proposed as a promising therapeutic strategy to promote neurogenesis following transplantation post SCI. In this study the antigenic and morphological phenotype of astrocytes, and the pattern of galectin expression was examined following SCI. Female Sprague Dawley rats were contused at thoracic (T) level T9 and treated with Cyclosporin-A (CsA) and/or NSCs 1 week post-injury. CsA and NSC treatment led to increased glial fibrillary acidic protein (GFAP), brain lipid binding protein (BLBP) and vimentin expression. Quiescent and reactive astrocytes were the most abundant cell morphology observed. Astrocytes co-localised with galectin-1 (Gal-1) and galectin-3 (Gal-3). Gal-1 expression was increased in both CsA and CSA + NSC treated animals 2 weeks post-treatment. Treatment with CsA and/or NSCs offers many prospective beneficial effects. A detailed revelation of how astrocytes are heterogeneously responsive to SCI, in addition to their affinity to galectins and response to CsA and NSCs is shown in this study. The current study’s findings support the potential application of NSCs as a novel therapeutic tool in the treatment of SCI.

Articular cartilage has only very limited regenerative capacities in humans. Tissue engineering techniques for cartilage damage repair are limited in the production of hyaline cartilage. Mesenchymal stem/stromal cells (MSCs) are multipotent stem cells and can be differentiated into mature cartilage cells, chondrocytes, which could be used for repairing damaged cartilage. Chondrogenesis is a highly complex, relatively inefficient process lasting over 3 weeks in vitro. In order to better understand chondrogenic differentiation, especially the commitment phase, we have performed transcriptional profiling of MSC differentiation into chondrocytes from early timepoints starting 15 minutes after induction to 16 hours and fully differentiated chondrocytes at 21 days. Transcriptional gene regulatory networks (GRN) were identified using time-course clustering and upstream-regulator predictions which revealed that cells start epigenetic reprogramming as early as 2 hours after induction and commit to differentiation within 4-6 hours. After that they adapt their gene expression to cater for differentiation specific protein production. These results suggest that interventions to improve the frequency and efficiency of differentiation should target early processes.

Chronic diseases characterized by bone and cartilage loss are associated with a reduced ability of progenitor cells to regenerate new tissues in an inflammatory environment. A promising strategy to treat such diseases is based on tissue repair mediated by human mesenchymal stem cells (hMSCs), but therapeutic outcomes are hindered by the absence of small molecules to efficiently modulate cell behaviour. Here, we applied a high-throughput drug screening technology to bioprospect a large library of extracts from Irish deep-sea organisms to induce hMSC differentiation toward musculoskeletal lineages and reduce inflammation of activated macrophages. The library included extracts from deep-sea corals, sponges and filamentous fungi representing a novel source of compounds for the targeted bioactivity. A validated hit rate of 3.4% was recorded from the invertebrate library, with cold water sea pens (octocoral order Pennatulacea), such as Kophobelemnon sp. and Anthoptilum sp., showing the most promising results in influencing stem cell differentiation toward osteogenic and chondrogenic lineages. Extracts obtained from deep-sea fungi showed no effects on stem cell differentiation, but a 6.8% hit rate in reducing the inflammation of activated macrophages. Our results demonstrate the potential of deep-sea organisms to synthetize pro-differentiation and immunomodulatory compounds that may represent potential drug development candidates to treat chronic musculoskeletal diseases.

Fungi populate deep Oceans in extreme habitats characterized by high hydrostatic pressure, low temperature and absence of sunlight. Marine fungi are potential major contributors to biogeochemical events, critical for marine communities and food web equilibrium under climate change conditions and a valuable source of novel extremozymes and small molecules. Despite their ecophysiological and biotechnological relevance, fungal deep-sea biodiversity has not yet been thoroughly characterized. In this study, we describe the culturable mycobiota associated with the deepest margin of the European Western Continental Shelf: sediments sampled at the Porcupine Bank and deep-water corals and sponges sampled in the Whittard Canyon. Eighty-seven strains were isolated, belonging to 43 taxa and mainly Ascomycota. Ten species and four genera were detected for the first time in the marine environment and a possible new species of Arachnomyces was isolated from sediments. The genera Cladosporium and Penicillium were the most frequent and detected on both substrates, followed by Candida and Emericellopsis. Our results showed two different fungal communities: sediment-associated taxa which were predominantly saprotrophic and animal-associated taxa which were predominantly symbiotic. This survey supports selective fungal biodiversity in the deep North Atlantic, encouraging further mycological studies on cold water coral gardens, often overexploited marine habitats.

Cell-based therapies, defined here as the delivery of cells in vivo to treat disease, have recently gained increasing public attention as a potentially promising approach to restore structure and function to musculoskeletal tissues. Although cell-based therapy has the potential to improve the treatment of disorders of the musculoskeletal system, there is also the possibility of misuse and misrepresentation of the efficacy of such treatments. The medical literature contains anecdotal reports and research studies, along with web-based marketing and patient testimonials supporting cell-based therapy. Both the American Society for Bone and Mineral Research (ASBMR) and the Orthopaedic Research Society (ORS) are committed to ensuring that the potential of cell-based therapies is realized through rigorous, reproducible, and clinically meaningful scientific discovery. The two organizations convened a multidisciplinary and international Task Force composed of physicians, surgeons, and scientists who are recognized experts in the development and use of cell-based therapies. The Task Force was charged with defining the state-of-the art in cell-based therapies and identifying the gaps in knowledge and methodologies that should guide the research agenda. The efforts of this Task Force are designed to provide researchers and clinicians with a better understanding of the current state of the science and research needed to advance the study and use of cell-based therapies for skeletal tissues. The design and implementation of rigorous, thorough protocols will be critical to leveraging these innovative treatments and optimizing clinical and functional patient outcomes. In addition to providing specific recommendations and ethical considerations for preclinical and clinical investigations, this report concludes with an outline to address knowledge gaps in how to determine the cell autonomous and nonautonomous effects of a donor population used for bone regeneration. © 2020 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 38:485-502, 2020.

International audience; The Signature Series Symposium "Cellular Therapies for Orthopaedics and Musculoskeletal Disease Proven and Unproven Therapies-Promise, Facts and Fantasy" was held as a pre-meeting of the 26th International Society for Cellular Therapy (ISCT) annual congress in Montreal, Canada, May 2, 2018. This was the first ISCT program that was entirely dedicated to the advancement of cell-based therapies for musculoskeletal diseases. Cellular therapies in musculoskeletal medicine are a source of great promise and opportunity. They are also the source of public controversy, confusion and misinformation. Patients, clinicians, scientists, industry and government share a commitment to clear communication and responsible development of the field. Therefore, this symposium convened thought leaders from around the world in a forum designed to catalyze communication and collaboration to bring the greatest possible innovation and value to patients with musculoskeletal conditions.

The modulation of expression levels of fluorescent fusion proteins (FFPs) is central for recombinant DNA technologies in modern biology as overexpression of proteins contributes to artifacts in biological experiments. In addition, some microscopy techniques such as fluorescence correlation spectroscopy (FCS) and single-molecule-based techniques are very sensitive to high expression levels of FFPs. To reduce the levels of recombinant protein expression in comparison with the commonly used, very strong CMV promoter, the herpes simplex virus thymidine kinase (TK) gene promoter, and mutants thereof were analyzed. Deletion mutants of the TK promoter were constructed and introduced into the Gateway (R) system for ectopic expression of enhanced green fluorescent protein (eGFP), monomeric cherry (mCherry), and FFPs containing these FPs. Two promoter constructs, TK2ST and TKTSC, were established, which have optimal low expression levels suitable for FCS studies in U2OS, HeLa CCL2, NIH 3T3, and BALB/c cells. Interestingly, when tested in these four cell lines, promoter constructs having a deletion within TK gene 5'-UTR showed significantly higher protein expression levels than the equivalent constructs lacking this deletion. This suggests that a negative regulatory element is localized within the TK gene 5'-UTR. Science Foundation Ireland. Grant Number: 06/CE/B1129 Else Kröner‐Fresenius‐Stiftung. Grant Number: 2013_A215 peer-reviewed

Background & Aim Induced pluripotent stem cells (iPSCs) have been considered as a protential alternative source for generating mesenchymal stem cells (MSCs) to meet the increased cell demands for both research and therapeutic applications. Based on the minimal criteria for defining MSCs published by the International Society for Cellular Therapy (ISCT standard), several different protocols have claimed success in differentiating MSCs from iPSCs (iMSCs). However, questions have arisen relating to the phenotypic fidelity of iMSCs because, irrespective of the derivation methods used, the cells generally have less capacity for adipo- and chondro-genesis when compared with primary MSCs. The objective of this study was to understand the cellular mechanism behind these discrepancies. Methods, Results & Conclusion iMSCs were derived using the embryoid body-based outgrowth method. We compared the differentiation ability, immunophenotype and gene expression profiles (GEPs) between multiple iMSC and BM-MSC lines. In addition, the impact of culture expansion on cell state was evaluated. The results showed that iMSCs met the ISCT standard but displayed significantly negligible adipogenic and chondrogenic capacity compared to BM-MSCs. GEPs analysis revealed that iMSCs expressed very high levels of vascular progenitor cell (VPC) genes (KDR and MSX2), whereas BM-MSCs had significantly greater levels of paraxial mesodermal genes (PDGFRα and MESP2). These distinct GEPs were maintained during culture expansion, indicating that, unlike BM-MSCs, iMSCs were more closely related to VPCs. Interestingly, the revealed characteristics of iMSCs corresponded to the known cell property of VPCs, where the later one displays partially overlapped immunophenotype as primary MSCs. However, although VPCs have osteo-, adipo- and chondro-genic potential, which cannot be fully activated in primary MSCs preferred differentiation condition. In summary, our results suggest a lineage misidentification of iMSCs, indicate the inadequacy of using the ISCT standard to distinguish different mesodermal progenitors and emphasize a necessity to validate the iMSCs identity with various lineage markers.

Background: With the advancement of cell therapy research, there is an increasing need for healthy volunteers (HV) to donate small volumes (30 ml) of human bone marrow (BM). The BM procedure required to procure small volumes is invasive, although short-lived (25 seconds), is not without risk. To ensure a sustainable supply of BM for research and cell therapy, greater information of the risks and factors that motivate HV to donate small volumes of BM will help optimize the procedure and HV enrolment, ensuring donors are fully informed of the potential risks. Objective: To identify the adverse events (AE) experienced by HV during and after small volume BM procedure and understand the motivating factors that infl uence HV to donate BM for research. Method: HV (n = 55) who donated BM (30 ml) for scientifi c research and provided informed consent were administered a questionnaire to identify the type, duration and severity of AE experienced during and post-BM aspiration; and to determine the motivating factors that influenced their willingness to donate BM. Results: Pain was experienced by 89% of participants during the BM procedure with moderate grade reported by 40%. One/more of the following AE were experienced by 73% of the volunteers post-BM procedure: pain, fatigue, site reaction, nausea and transient hypotension. AE resolved within an average of three days. The reported motivational factors ranked in the following order:first, to advance research for the benefit of future patients; compensation for participation; free medical check-up; lastly, the research question was interesting. Conclusion: Young HV, motivated primarily by altruism and financial compensation, risk the occurrence of transient AE following donation of small-volume BM for research. Not

Background Mesenchymal stromal cells (MSCs) demonstrate considerable promise for acute respiratory distress syndrome (ARDS) and sepsis. However, standard approaches to MSC isolation generate highly heterogeneous cell populations, while bone marrow (BM) constitutes a limited and difficult to access MSC source. Furthermore, a range of cell manufacturing considerations and clinical setting practicalities remain to be explored. Methods Adult male rats were subject to E. coli-induced pneumonia and administered CD362+ umbilical cord (UC)-hMSCs using a variety of cell production and clinical relevance considerations. In series 1, animals were instilled with E. coli and randomized to receive heterogeneous BM or UC-hMSCs or CD362+ UC-hMSCs. Subsequent series examined the impact of concomitant antibiotic therapy, MSC therapeutic cryopreservation (cryopreserved vs fresh CD362+ UC-hMSCs), impact of cell passage on efficacy (passages 3 vs 5 vs 7 vs 10), and delay of administration of cell therapy (0 h vs 6 h post-injury vs 6 h + 12 h) following E. coli installation. Results CD362+ UC-hMSCs were as effective as heterogonous MSCs in reducing E. coli-induced acute lung injury, improving oxygenation, decreasing bacterial load, reducing histologic injury, and ameliorating inflammatory marker levels. Cryopreserved CD362+ UC-hMSCs recapitulated this efficacy, attenuating E. coli-induced injury, but therapeutic relevance did not extend beyond passage 3 for all indices. CD362+ UC-hMSCs maintained efficacy in the presence of antibiotic therapy and rescued the animal from E. coli injury when delivered at 6 h + 12 h, following E. coli instillation. Conclusions These translational studies demonstrated the efficacy of CD362+ UC-hMSCs, where they decreased the severity of E. coli-induced pneumonia, maintained efficacy following cryopreservation, were more effective at early passage, were effective in the presence of antibiotic therapy, and could continue to provide benefit at later time points following E. coli injury.

Articular cartilage lacks an intrinsic repair capacity and due to the ability of mesenchymal stem cells (MSCs) to differentiate into chondrocytes, MSCs have been touted as a cellular source to regenerate damaged cartilage. However, a number of prevailing concerns for such a treatment remain. Generally, administration of MSCs into a cartilage defect results in poor regeneration of the damaged cartilage with the repaired cartilage consisting primarily of fibro-cartilage rather than hyaline cartilage. Methods that improve the chondrogenic potential of transplanted MSCs in vivo may be advantageous. In addition, the proclivity of MSC-derived cartilage to undergo hypertrophic differentiation or form bone in vivo also remains a clinical concern. If MSC-derived cartilage was to undergo hypertrophic differentiation in vivo, this would be deleterious in a clinical setting. This study focuses on establishing a mechanism of action by which hypoxia or low oxygen tension can be used to both enhance chondrogenesis and attenuate hypertrophic differentiation of both MSC and ATDC5 derived chondrocytes. Having elucidated a novel mechanism of action, the subsequent goals of this study were to develop an in vitro culture regime to mimic the beneficial effects of physiological low oxygen tension in a normoxic environment.

Mesenchymal stromal cells (MSCs) have firmly occupied the attention of orthopedic clinicians and scientists for most of the last 25 years. Hundreds of laboratories worldwide have carried out research aimed at unraveling the biological characteristics of these cells and probing the manner in which they potentially contribute to cartilage and bone repair. Clinical trials registries indicate that they are also being tested in patient studies for a wide range of conditions such as osteoarthritis, rheumatoid arthritis, fracture repair, regeneration of articular cartilage, tendon repair, and for treatment of degenerative disc disease. Despite these efforts, the effectiveness of MSCs as a treatment modality for these conditions is still uncertain and market authorizations have been limited. In addition, critical and clear phenotypic parameters for defining MSCs are uncertain and a coherent biological framework surrounding the therapeutic mechanism of action is not yet available. Added to this, cell manufacturing protocols are complex and costly and present substantial challenges in terms of regulatory oversight and standardization. Despite these obstacles, MSCs still remain at the forefront of efforts in Regenerative Medicine, based on a conviction that this technology can provide an effective treatment paradigm for major diseases where there is still an unmet need. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:1229-1235, 2019.

The Committee on Industrial Organisation was established in the early 1960s to evaluate the level of preparedness of Irish industry for the imminent dismantling of the country's protectionist trade barriers. The Committee's sectoral reports list the 900 or so manufacturing firms that it surveyed and that together accounted for more than half of Irish manufacturing employment. From a range of archival sources, this article identifies the nationality of ownership of most of these firms, alongside their date of establishment and level of employment in 1960. Industrial grants data are used to strip out export-oriented businesses that began to arrive in the 1950s. This makes it possible to estimate the employment share and sectoral presence of tariff-jumping foreign firms. The latter is analysed through the lens of modern perspectives on foreign direct investment. The article also enhances our understanding of the interest-group politics of the trade liberalization process.

Familial osteochondritis dissecans (FOCD) is an inherited skeletal defect characterized by the development of large cartilage lesions in multiple joints, short stature, and early onset of severe osteoarthritis. It is associated with a heterozygous mutation in the ACAN gene, resulting in a Val-Met replacement in the C-type lectin domain of aggrecan. To understand the cellular pathogenesis of this condition, we studied the chondrogenic differentiation of patient bone marrow mesenchymal stromal cells (BM-MSCs). We also looked at cartilage derived from induced pluripotent stem cells (iPSCs) generated from patient fibroblasts. Our results revealed several characteristics of the differentiated chondrocytes that help to explain the disease phenotype and susceptibility to cartilage injury. First, patient chondrogenic pellets had poor structural integrity but were rich in glycosaminoglycan. Second, it was evident that large amounts of aggrecan accumulated within the endoplasmic reticulum of chondrocytes differentiated from both BM-MSCs and iPSCs. In turn, there was a marked absence of aggrecan in the extracellular matrix. Third, it was evident that matrix synthesis and assembly were globally dysregulated. These results highlight some of the abnormal aspects of chondrogenesis in these patient cells and help to explain the underlying cellular pathology. The results suggest that FOCD is a chondrocyte aggrecanosis with associated matrix dysregulation. The work provides a new in vitro model of osteoarthritis and cartilage degeneration based on the use of iPSCs and highlights how insights into disease phenotype and pathogenesis can be uncovered by studying differentiation of patient stem cells. We sincerely thank the family members who participated in the study and Dr. Yelverton Tegner, Luleå Technical University, Luleå, Sweden, for providing skin samples. We acknowledge the facilities and technical assistance of the Flow Cytometry Facility at the National University of Ireland Galway, a facility that is funded by National University of Ireland Galway and the Irish Government's Programme for Research in Third Level Institutions, Cycle5, National Development Plan 2007–2013. We thank Pierce Lalor, Dr. Kerry Thompson, and the Centre for Microscopy and Imaging (http://www.imaging.nuigalway.ie) for assistance with TEM and confocal imaging, and Jingqiu Zhang (A*STAR Institute of Medical Biology, Singapore) for assistance with the iPSC reprogramming technique. We also thank Maggie Hall for helpful editing of the manuscript. This study was funded by Science Foundation Ireland Grant SFI 09/SRC.B1794 and Irish Research Council Grant GOIPG/2014/96. This project has also received funding from the European Union's Seventh Framework Programme for Research, Technological Development and Demonstration under grant agreement no. 223298. peer-reviewed

Advanced therapeutics, specifically cell and gene therapies, provide an opportunity to target previously unmet clinical conditions and offer a potential solution to the social and economic burden associated with an aging population [1,2]. Human mesenchymal stem/stromal cells (hMSCs) are a promising cell therapy candidate for the treatment of numerous clinical indications [3]; however, a transformation is required in the way we isolate, manufacture, characterize and deliver these therapies to ensure they are both efficacious and affordable [2,4]. Manufacture of hMSCs requires in vitro expansion to increase the available number of cells to meet clinical demand; however, progress is impeded by the lack of advanced methods for the isolation and expansion of cells that are scalable, amenable for automation and closed. Many allogeneic processes still require manual intervention which has significant quality and cost implications [5], and developing reproducible, consistent bioprocesses is still a major challenge. Even when a scale-out approach is to be employed (e.g., for autologous therapies), there is a significant practical challenge of manipulating, processing and segregating multiple production batches in an aseptic, closed manner. Such processes demand small units for manufacture in which line segregation is a priority in order to avoid product–patient mismatch and disease transmission. As such, there is an industrial trend toward automated systems as it is increasingly recognized that such systems facilitate consistent manufacture and will play a pivotal role in the translation of cell therapies by improving quality control, process economics, scalability, process capability and provide a platform for understanding process variation and optimization [6]. Through AUTOSTEM (European Commission Horizon 2020 funded research and innovation actions), academic and industrial groups from across the EU are working in collaboration to take a holistic approach to enable large-scale hMSC production, at clinical-grade quality, by implementing a robotic automated pipeline for cell isolation and culture [7]. AUTOSTEM builds on the ground-breaking work of the StemCellFactory project [8] which built and demonstrated an automated robotic pipeline for the production of human induced pluripotent cell lines (Figure 1). AUTOSTEM develops this technology further to enable automated, closed and good manufacturing practice (GMP)ready hMSCs (a ‘StromalCellFactory’) and will focus on the development of

Cell-based therapies, defined here as the delivery of cells in vivo to treat disease, have recently gained increasing public attention as a potentially promising approach to restore structure and function to musculoskeletal tissues. Although cell-based therapy has the potential to improve the treatment of disorders of the musculoskeletal system, there is also the possibility of misuse and misrepresentation of the efficacy of such treatments. The medical literature contains anecdotal reports and research studies, along with web-based marketing and patient testimonials supporting cell-based therapy. Both the American Society for Bone and Mineral Research (ASBMR) and the Orthopaedic Research Society (ORS) are committed to ensuring that the potential of cell-based therapies is realized through rigorous, reproducible, and clinically meaningful scientific discovery. The two organizations convened a multidisciplinary and international Task Force composed of physicians, surgeons, and scientists who are recognized experts in the development and use of cell-based therapies. The Task Force was charged with defining the state-of-the art in cell-based therapies and identifying the gaps in knowledge and methodologies that should guide the research agenda. The efforts of this Task Force are designed to provide researchers and clinicians with a better understanding of the current state of the science and research needed to advance the study and use of cell-based therapies for skeletal tissues. The design and implementation of rigorous, thorough protocols will be critical to leveraging these innovative treatments and optimizing clinical and functional patient outcomes. In addition to providing specific recommendations and ethical considerations for preclinical and clinical investigations, this report concludes with an outline to address knowledge gaps in how to determine the cell autonomous and nonautonomous effects of a donor population used for bone regeneration. © 2019 American Society for Bone and Mineral Research.

Background & Aim Bone marrow-derived mesenchymal stromal cells (BM-MSCs) are an attractive candidate for use in regenerative therapies and are being widely investigated for the treatment of a broad spectrum of diseases. Despite this, the efficacy of treatment is unproven and interpretation of trial data is difficult, in part due to the heterogeneity of MSC preparations as a result of variable manufacturing processes. MSCs are currently isolated in basal media containing fetal bovine serum (FBS), platelet lysate or other supplements but little is understood about the impact of changes in media composition on the biology, therapeutic action or potency of MSCs. The aim of this study was to analyse the influence of widely used culture media on the growth parameters, differentiation potential and surface marker expression of human BM-MSCs. Methods, Results & Conclusion Whole bone marrow obtained from the iliac crest of a 24-year-old healthy donor was plated in a standard basal medium with 5 different supplements: (1) a proprietary serum-free supplement, (2) 5% human platelet lysate, (3) 10% FBS, screened and proven to support MSC isolation and expansion (4) 10% screened FBS and 1 ng/ml recombinant human basic fibroblast growth factor (FGF2) and (5) 10% unscreened FBS and 1 ng/ml FGF2. The impact of media composition on growth kinetics, morphology, differentiation propensity and surface immunophenotype was assessed. The latter involved the BD Lyoplate flow cytometry screening panel of 242 monoclonal antibodies. MSC growth kinetics were highly influenced by expansion medium, with the greatest proliferation rate observed in MSCs cultured in the serum-free supplemented medium. Cell morphology varied greatly between cell preparations. Adipogenesis and osteogenesis were maintained in all culture conditions, but the degree of differentiation varied between conditions. Chondrogenesis was most starkly effected by changes in expansion medium and was only observed in serum-containing media. Analysis of surface marker expression revealed significant variation in response to culture conditions, with 70 of the 242 markers differentially expressed between cell preparations. These data suggest that changes in the composition of the culture media – for example switching batches of FBS – have profound effects on key biological attributes of BM-MSCs and on their surface marker expression profiles. This demonstrates the critical need for harmonisation of media formulation to aid interpretation of clinical results.

Foreign direct investment (FDI) statistics are widely used to study the impact of international capital movements and multinational enterprise (MNE) activities. FDI intensity is also an important indicator of globalisation and economic integration. Datasets spanning long time periods and with broad country coverage have been employed in numerous studies to analyse various aspects of the determinants and consequences of FDI. Focusing on a relatively homogeneous group of six Western European EU countries, the present study finds major inconsistencies in the construction and coverage of these data both through time and across countries, leading to large discrepancies. Asymmetries will be far greater for broader groups of more economically and institutionally diverse countries. This study recommends extreme caution in drawing conclusions based on FDI data.

Mesenchymal stromal cell (MSC) therapies have been pursued for a broad spectrum of indications but mixed reports on clinical efficacy have given rise to some degree of skepticism regarding the effectiveness of this approach. However, recent reports of successful clinical outcomes and regulatory approvals for graft-versus-host disease, Crohn's disease and critical limb ischemia have prompted a shift in this perspective. With hundreds of clinical trials involving MSCs currently underway and an increasing demand for large-scale manufacturing protocols, there is a critical need to develop standards that can be applied to processing methods and to establish consensus assays for both MSC processing control and MSC product release. Reference materials and validated, uniformly applied tests for quality control of MSC products are needed. Here, we review recent developments in MSC manufacturing technologies, release testing and potency assays. We conclude that, although MSCs hold considerable promise clinically, economies of scale have yet to be achieved although numerous bioreactor technologies for scalable production of MSCs exist. Additionally, rigorous disease-specific product testing and comprehensive understanding of mechanisms of action, which are linked to relevant process and product release potency assays, will be required to ensure that these therapies continue to be successful.

Osteoarthritis (OA) is an inflammatory condition still lacking effective treatments. Mesenchymal stem/stromal cells (MSCs) have been successfully employed in pre-clinical models aiming to resurface the degenerated cartilage. In early-phase clinical trials, intra-articular (IA) administration of MSCs leads to pain reduction and cartilage protection or healing. However, the consistent lack of engraftment indicates that the observed effect is delivered through a “hit-and-run” mechanism, by a temporal release of paracrine molecules. MSCs express a variety of chemokines and cytokines that aid in repair of degraded tissue, restoration of normal tissue metabolism and, most importantly, counteracting inflammation. Secretion of therapeutic factors is increased upon licensing by inflammatory signals or apoptosis, induced by the host immune system. Trophic effectors are released as soluble molecules or carried by extracellular vesicles (ECVs). This review provides an overview of the functions and mechanisms of MSC-secreted molecules found to be upregulated in models of OA, whether using in vitro or in vivo models.

What We Already Know about This TopicWhat This Article Tells Us That Is NewBackgroundHuman mesenchymal stromal cells demonstrate promise for acute respiratory distress syndrome, but current studies use highly heterogenous cell populations. We hypothesized that a syndecan 2 (CD362)–expressing human mesenchymal stromal cell subpopulation would attenuate Escherichia coli–induced lung injury and enhance resolution after ventilator-induced lung injury.MethodsIn vitro studies determined whether CD362+ human mesenchymal stromal cells could modulate pulmonary epithelial inflammation, wound healing, and macrophage phagocytosis. Two in vivo rodent studies determined whether CD362+ human mesenchymal stromal cells attenuated Escherichia coli–induced lung injury (n = 10/group) and enhanced resolution of ventilation-induced injury (n = 10/group).ResultsCD362+ human mesenchymal stromal cells attenuated cytokine-induced epithelial nuclear factor kappa B activation, increased epithelial wound closure, and increased macrophage phagocytosis in vitro. CD362+ human mesenchymal stromal cells attenuated Escherichia coli–induced injury in rodents, improving arterial oxygenation (mean ± SD, 83 ± 9 vs. 60 ± 8 mmHg, P < 0.05), improving lung compliance (mean ± SD: 0.66 ± 0.08 vs. 0.53 ± 0.09 ml · cm H2O−1, P < 0.05), reducing bacterial load (median [interquartile range], 1,895 [100–3,300] vs. 8,195 [4,260–8,690] colony-forming units, P < 0.05), and decreasing structural injury compared with vehicle. CD362+ human mesenchymal stromal cells were more effective than CD362− human mesenchymal stromal cells and comparable to heterogenous human mesenchymal stromal cells. CD362+ human mesenchymal stromal cells enhanced resolution after ventilator-induced lung injury in rodents, restoring arterial oxygenation (mean ± SD: 113 ± 11 vs. 89 ± 11 mmHg, P < 0.05) and lung static compliance (mean ± SD: 0.74 ± 0.07 vs. 0.45 ± 0.07 ml · cm H2O−1, P < 0.05), resolving lung inflammation, and restoring histologic structure compared with vehicle. CD362+ human mesenchymal stromal cells efficacy was at least comparable to heterogenous human mesenchymal stromal cells.ConclusionsA CD362+ human mesenchymal stromal cell population decreased Escherichia coli–induced pneumonia severity and enhanced recovery after ventilator-induced lung injury.