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Sensorineural hearing loss (SNHL) is commonly caused by the death or dysfunction of cochlear cell types as a result of their lack of regenerative capacity. However, regenerative medicine, such as stem cell therapy, has become a promising tool to cure many diseases, including hearing loss. In this study, we determined whether DPSCs could differentiate into cochlear hair cell in vitro. DPSCs derived from human third molar dental pulp were induced into NSCs using a medium containing basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) for 7 days, and then into cochlear hair cell using a medium containing EGF and IGF-1 for the next 14 days. We used the neuroepithelial protein marker nestin and cochlear hair cell marker myosin VIIa as the markers for cells differentiation. Cells expressing the positive markers under the microscope were confirmed to have differentiated into cochlear hair cell. DPSCs were successfully induced to differentiate into NSCs, with mean 24% nestin-positive cells. We found that DPSC-derived NSCs have a great capacity in differentiating into inner ear hair cell-like cells with an average of 81% cells presenting myosin VIIa. Thus, DPSCs have high potential to serve as a good resource for SNHL treatment. We found the high potential of DPSCs to differentiate into NSC. The ability of DPSCs in differentiating into neural lineage cell made them a good candidate for regenerative therapy in neural diseases, such as SNHL

The COVID-19 disease, which is caused by the novel coronavirus, SARS-CoV-2, has affected the world by increasing the mortality rate in 2020. Currently, there is no definite treatment for COVID-19 patients. Several clinical trials have been proposed to overcome this disease and many are still under investigation. In this review, we will be focusing on the potency of mesenchymal stem cells (MSCs) and MSC-derived secretome for treating COVID-19 patients. Fever, cough, headache, dizziness, and fatigue are the common clinical manifestations in COVID-19 patients. In mild and severe cases, cytokines are released hyper-actively which causes a cytokine storm leading to acute respiratory distress syndrome (ARDS). In order to maintain the lung microenvironment in COVID-19 patients, MSCs are used as cell-based therapy approaches as they can act as cell managers which accelerate the immune system to prevent the cytokine storm and promote endogenous repair. Besides, MSCs have shown minimal expression of ACE2 or TMPRSS2, and hence, MSCs are free from SARS-CoV-2 infection. Numerous clinical studies have started worldwide and demonstrated that MSCs have great potential for ARDS treatment in COVID-19 patients. Preliminary data have shown that MSCs and MSC-derived secretome appear to be promising in the treatment of COVID-19. Lay Summary The COVID-19 disease is an infection disease which affects the world in 2020. Currently, there is no definite treatment for COVID-19 patients. However, several clinical trials have been proposed to overcome this disease and one of them is using mesenchymal stem cells (MSCs) and MSC-derived secretome for treating COVID-19 patients. During the infection, cytokines are released hyper-actively which causes a cytokine storm. MSCs play an important role in maintaining the lung microenvironment in COVID-19 patients. They can act as cell managers which accelerate the immune system to prevent the cytokine storm and promote the endogenous repair. Therefore, it is important to explore the clinical trial in the world for treating the COVID-19 disease using MSCs and MSC-derived secretome.

Background: Retinitis Pigmentosa (RP) is a rare retinal genetic disease without available treatment to date. Previous studies show growth factor and regenerative ability of secretome from cultured allogeneic umbilical chord mesenchymal stem cells (UC-MSC). This studyaimed to report multifocal electroretinography of retinal photoreceptor responsefrom RP patients, before and after injection of secretome from allogeneicUC MSC.Patients and Methods: Four subjects with severely damage retina (visual field defect of 25 %- 50% in initial Humphrey perimetry examination) were recruited and givenperibulbar injection of secretome from allogeneic cultured UC-MSC. Visual acuity, visual field examination, multifocal electroretinogram of retinal photoreceptor examination were observed before and periodically after injection until six months periodResults: Overall, we observed subtle changes in N1 and P1 amplitude and implicit time only in ring 1 at 1, 3 and 6 month post secretome injection. Conclusion: Peribulbar injection of allogeneic UC-MSC secretome had very miminal influence on photoreceptor activity in late-stage RP patients.

BACKGROUND: Type 2 Diabetes Mellitus (T2DM) is a metabolic disease, due to the disorder of insulin function, insulin secretion, or both. Long-term hyperglycemia conditions promote endothelial dysfunction precedes to the development of multiple organ dysfunctions. Endothelial progenitor cells (EPCs) and hematopoietic stem cell (HSCs) are the key cellular effectors of postnatal neovascularization and play central role in endothelial dysfunction. However, in T2DM condition, the number of apoptotic HSCs increase, it may cause the reduction in potency and number of EPCs. In diabetes, the circulating EPCs number decrease and their functionality is impaired, but mechanism underlie of this impairement is unknown. The purpose of this study was to examine the relationship duration diabetes with the number and potency of EPC cells in T2DM patients controlled and poorly controlled.METHODS: Thirty-eight T2DM male patients were classified into two group based on Indonesian Society of Endocrinology/Perkumpulan Endokrinologi Indonesia (PERKENI) criteria on T2DM. The first group was a controlled glycemic condition group (hemoglobin A1c (HbA1C) 7.0%). Cluster of differentiation (CD)34+ and CD133+ expressions were used as specific marker for EPC, while quantified bright aldehyde dehydrogenase (ALDHbr) assay was used to represented the potency of EPCs.RESULTS: This study showed that in poorly controlled T2DM group the number of EPCs was lower by 24.80% (p

BACKGROUND: Umbilical Cord Blood (UCB) has been widely use as regenerative medicine due to the content of undifferentiated cell which have capability to do self-renewal and differentiation into various type of cell called stem cells. Recent studies show that UCB contains not only hematopoietic stem cell (HSC) but also others stem cell and progenitor cell such as endothelial progenitor cell (EPC) and very small embryonic-like stem cell (VSEL). It is beliefs that HSC and EPC shared the same progenitor. In this study, correlation between the cell number of HSC, EPC and VSEL is analyzed in umbilical cord blood as the source of stem cell for clinical application. METHODS: The cell number of HSC, EPC and VSEL is counted from cryopreserved UCB collected from 22 women delivered via cesarean section which already stored for more than 2 years in this study. Sample were incubated with antibodies such as cluster of differentiation (CD)34-phycoerythrin (PE)/CD45-fluorescein isothiocyanate (FITC), CD133/1 (anti-CD (AC)133)-antigen-presenting cell (APC) and, CD184 (C-X-C chemokine receptor (CXCR)4)-PE.Vio770 to detect the present of HSC, EPC and VSEL in UCB. Sample were analyze using flowcytometer BD FACS Canto II. RESULTS: The cell population of HSC and late-EPC is 0.009% and 0.01% of total cell in UCB. VSEL only represented 0.001% from total cell in UCB, showing the lowest number of cell population in UCB. The correlation between the cell number of HSC and EPC is r=0,483*, p =0.023) and between HSC and VSEL is r=0.510*, p =0.015. CONCLUSION: In this study, both EPC and VSEL have a significant positive correlation with HSC. KEYWORDS: stem cell, umbilical cord blood, endothelial progenitor, flowcytometry

Background & Aim Background Stem cell therapy has been considered as a potential strategy in regenerative for the patient with neurological deficits. Mesenchymal Stem/Stromal Cells (MSC) have been extensively investigated as a treatment in stroke. However, the MSC lost its potency for neural repair in aged patients due to the limited numbers of Neural Stem Cell (NSC). NSC is essential for neurogenesis of the injured brain. In vitro studies have shown that MSCs which were cultured in the laboratory has the capability to transform into different cells. Adding stimulants such as growth factors into the culture media could induce the trans-differentiation of MSC into NSC. However, to produce NSC in the clinical setting on a large scale is still a challenge. Objective In this study, we like to evaluate the use of epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), heregulin B27, and N2 the trans-differentiation mechanism of umbilical cord MSC (UC-MSC) into NSC. Methods, Results & Conclusion Method MSC was transferred into a 24-well plate at 2 × 104 cells per well. Induction of NSC is done using DMEM containing N2/B27 supplement, 20 ng/ml epidermal growth factor (EGF), and 10 ng/ml basic fibroblast growth factor (bFGF). Identification of success trans-differentiation is analyzed using NeuroFlour CDr3 antibody. Result and Discussion MSC transdifferentiates into NSC as an important source of cell in therapy for neurological diseases. UC-MSC was induced using N2 to helps the cell to commit to differentiation and its survival. B27 is used for the long term survival of neurons cells and helps the NSC to grow and proliferates. bFGF beliefs as pre-inductive molecules that enhance neuronal differentiation and self-renewal. Upon bFGF stimulation, NGFβR and NRP1 will upregulate and promotes differentiation. On the contrary, EGF induces differentiation at lower concentrations. EGF is used for inducing the expression of the neuron and glial marker. To confirm the successful transdifferentiating process, the expression of CDr3 in the cell culture is analyzed. CDr3 positive result as the membrane-permeable fluorescent probe was shown in Figure 1. NeuroFluor CDr3 binds to human fatty acid-binding protein 7 (FABP7) which use to confirm the neural induction from human stem cells. Conclusion UC-MSC could be cultured and differentiate into NSC in vitro when supplemented with N2/B27, bFGF, and EGF, which may produce NSC in the clinical setting on a large scale.

Background & Aim Umbilical cord tissue is a potential source of Mesenchymal Stem Cells (MSCs). These cells have been widely used especially in pre-clinical and clinical applications resulting its various effectiveness. Banking process of umbilical cord involves transportation from the hospital where the cord is received to the banking facility to be processed further. Prior to processing, pre-sterility test must be conducted and while waiting for the result, the umbilical cord is quarantined. This quarantine process has to be carried out carefully to maintain cell's recovery and viability. Suitable range of time should be chosen to deliver high quality cells. In this study, we aim to determine the maximum time for quarantined umbilical cord to be processed. Methods, Results & Conclusion One umbilical cord (UC2018030001) was used in this study. Medium transportation is used to carry the umbilical cord to banking facility. Prior to processing, pre sterility was done using spread plate method by spreading the medium transport onto TSA agar. After the results were out and passed, the tissues were put into quarantine medium and were processed at day 0, 7, 14, and 21 for MSCs isolation. The tissue was processed using explant method and sub-cultured until 3rd passage. Graphic shown below are the cell doubling level from different passages at various days of quarantine. The doubling level remain constant with maximum deviation at 0.512. Results showed that tissues which was quarantined until day 21 still produce cells normally. The cell number are slowly increasing because the higher the passage, the more pure the cells are so they can propagate easily. This shows normal growth of normal cells. Their doubling level are also on track with no significant difference. In brief, umbilical cord tissue can be quarantined until 21 days and still produce cells normally.

Background & Aim Coronary artery disease (CAD) is one of the leading cause of death in the world. Currently, the common therapy for this disease is to use drugs and surgical procedures however the result has been less optimum. Therefore, it is necessary to develop an alternative treatment using Mesenchymal Stem Cell (MSC) and Conditioned Medium (CM). Methods, Results & Conclusion MSCs were obtained from the umbilical cord of eligible donor and process in a cGMP-standardized laboratory certified by the Indonesian Ministry of Health. A 52-year-old subject who experienced Coronary Artery Disease was treated with PCI four years before the stem cell therapy. The subject signed an informed consent for stem cell therapy before the injection. Allogeneic UC-MSC for CAD was packed in a syringe containing 20 ml NaCl 0.9% with cell concentration of 20 × 106. Umbilical Cord-Mesenchymal Stem Cells were administered intravenously and conducted three times over 4 months period. Meanwhile, Conditioned Medium (CM) was applied intranasally with a dose of 0.4 cc/day for a week after each UC-MSCs administrated. Assessment with Magnetic Resonance Imaging (MRI) shows improvement on the Left Ventricular Ejection Fraction (LVEF) from 55% at baseline and 75% at 3 weeks after first stem cell transplantation. Endothelial Cell Progenitor (EPC) count show decreasing the number of EPC cells from 11,893 cells/mL into 4,932 cells/mL. This shows the angiogenesis process takes place after the UC-MSC injection. On the twelve weeks after the first injection, the subject undergo spirometry test and shows improvement in the vital capacity (85% into 94%), forced capacity (85% into 94%), and forced expansion volume (93% into 103%). There are no adverse events occur and the patient experience improvement in physic, MRI, laboratory and spirometry assessment.

Background & Aim Background: Type 2 diabetes mellitus (T2DM) is a metabolic disease caused by insulin disruption, insulin secretion or both. Hyperglycemia occurs in diabetes, where long-term hyperglycemia causes an increase in oxidative stress, a decrease in antioxidants and an increase in pro-inflammatory factors that contribute to endothelial dysfunction. In these conditions where microenvironment is not right, the amount of available ATP is not adequate, causing a decrease in the number and potential of Endothelial Progenitor Cells (EPC) in playing its function and role in the decreased colonization process. In this preliminary study, the aim was to prove changes in microenvironment by adding growth media to improve the condition of the microenvironment (niche) in an in-vitro able to increase the number and potential of EPC Methods, Results & Conclusion Method: Circulating EPC counts were calculated using a combination of expressions from CD34+ and CD133+ antigen surfaces using BD FACS Canto II flow cytometry, and the potential of EPC was done by adding ALDH bright staining. Then the EPC was isolated and cultured on the growth medium, then cultured for seven days. After that, the number of EPC circulating and the EPC potential is recalculated Result The results of this study indicate that after the addition of growth medium and incubated for seven days, the number of EPCs increased by 413.20%, and 1,401.1% in the controlled and uncontrolled group. Also, the potential of EPC showed by express of ALDHbright was obtained at the end of incubation of 99.9%, in each group in diabetes. Discussion This study illustrates that by adding a growth medium, it can restore microenvironment conditions to be better characterised by the ability of EPC to express bright ALDH which increases very high. Bright ALDH activity shows the ability to re-populating and differentiation and illustrates the operation of retinoids that have a pro-angiogenic effect with modulation of endothelial cell cytokines, the impact of migration and vasculogenesis. Conclusion The addition of growth medium in EPC culture from peripheral blood improves microenviral conditions and can increase the amount of EPC and the potential of EPC associated with its role in the colonisation function.

Background & Aim Umbilical Cord derived Mesenchymal Stem Cells (UC-MSCs) and Stromal Vascular Fraction (SVF) has been widely used in preclinical and clinical applications, with varying degrees of effectiveness (Menasche, 2011). Due to freshness condition and different cell doses for therapy of each patient, the laboratory have to provide cell product while maintaining the optimal quality of the cells. The effects of storage conditions such as storage solutions, temperature, and duration of cell storage greatly influence the cell quality for transplant. The quality of cells directly can be evaluated from the cell viability rapid measurement. High cell viability shows that proportion of live cells is higher than dead cells, while low cell viability indicates that many cells have begun to be critical or even death. Therefore, our research aims to evaluate of several mediums and temperatures storage to maintain stability of UC-MSCs and SVF viability for transplantation. Methods, Results & Conclusion Both of UC-MSCs and SVF were divided into 4 groups with two variables temperature and medium storage. Cells have been evaluated in room temperature (RT) (18-22°C) and cold temperature (2-8°C) with 0.9% NaCl and conditioned medium (UC-MSCs) or autologous serum (SVF). The viability from each group was assessed day by day using Tryphan Blue. At room temperature, viability of cells in both storage mediums ware significantly decreased while stored at cold temperature the viability was more stable. The highest viability of UC-MSCs at RT was obtained from cells in 0.9% NaCl solution (77% viable). However, the highest UC-MSCs viability at cold temperature was obtained from cell storaged in 0.9% NaCl solution (93% viable). Data shown, for SVF viability from day 0 (when SVF was produced) to day 6, cells are most stable in 0.9% NaCl and autologous serum suspension. Both of them which stored at cold temperature stable on 90.4% viability. SVF in 0.9% NaCl at room temperature has a less stable viability where cell viability decrease from 96% at first day until 77% at sixth day. Cold temperature (2-8°C) can maintain stability of UC-MSCs and SVF cell viability. The most stable of UC-MSCs viability was obtained from cell storage in 0.9% NaCl solution and SVF in autologous serum.

Background & Aim Adipose tissue provides an abundant source of stromal vascular fraction (SVF), which resides a potential regenerative medicine named mesenchymal stem cells (MSC). Storage period and temperature before isolation may impact the quality of isolated cells. Here, we identify the effect of storage periods and temperatures to SVF viability and cell number along with the MSC recovery. Methods, Results & Conclusion A total of 70 ml donor-adipose tissue was divided into two different groups, room temperature (RT) and 4-8°C. SVF was isolated using manual method. Each group was process after stored for 4, 5, 6, 7, and 12 days. After it passed each incubation time, adipose samples were processed enzymatically until SVF was obtained. The viability of SVF stored at RT was decreasing until 40%, but relatively constant when stored at 4-8°C (>75%). Nevertheless, the sample stored at RT revealed to have better proliferation and colonization than sample stored at 4-8°C. The sample retained the proliferation capability when stored at RT up to 7 days, but only up to 5 days when stored at 4-8°C. The average of SVF number was higher in the sample stored at RT (8,552,315 cell/mL) compared to the sample at 4-8°C (4,581,111 cell/mL). The highest SVF number was observed in sample stored at 4 days before isolation. The average of MSC recovery was also higher at RT compared to sample stored at 4-8°C, which was 6% and 3%, respectively. In conclusion, adipose tissue is preferable to be stored at RT before isolation for higher SVF total number and MSC recovery. The adipose tissue should be processed immadiately or stored only up to 4 days after sample collection.

Stroke is a leading cause of death and long-term disability. This due to the ischemic event that cause by embolism of blockage blood flow. Thrombolytic agent plasminogen activator (tPA) is the only treatment approved by FDA. However, the used of tPA is limited to the short time window period. Neural stem cells (NSCs) show the potential to repair neuronal damage naturally after stroke. However, isolating NSCs is a challenging process due to the limitations of the method and its invasiveness. Some studies that had used mesenchymal stem cell (MSCs) as the main source of stem cell for therapy show that MSCs have the potency to differentiate into NSCs. in vitro, a differentiation process from MSC to NSC has been developed by combining the supplement or growth factor needed in the culture media.Keywords: stem cells, neuron stem cell, mesenchymal stem cell, stroke, trans-differentiation

BACKGROUND: Central obesity is closely associated with chronic inflammation, characterized by abnormal cytokine production such as IL-1β, tumor necrosis factor-α (TNF-α) and tumor necrosis factor receptor-2 (TNFR-2). Central obesity and chronic inflammation form a complex link with insulin resistance, leading to the development of type-2 diabetes mellitus (T2DM) and cardiovascular disease (CVD). Knowing the coinciding occurrence of chronic inflammation and central obesity, this study aimed to examine pro-inflammatory cytokine profiles of Indonesian adult men with central obesity.METHODS: This cross-sectional study recruited 80 apparently healthy Indonesian adult men, aged 23-53 years with waist circumference of 64-125 cm. This study was done in Jakarta. Measurements included clinical parameters like systolic blood pressure (SBP), diastolic blood pressure (DBP), fasting blood glucose (FBG), aspartate amino transferase (AST), alanine amino transferase (ALT), creatinine, high sensitivity C-reactive protein (hsCRP), anthropometric parameters, namely weight, height and waist circumference; and pro-inflammatory cytokines TNF-α, soluble TNFR-2 (sTNFR-2) and IL-1β.RESULTS: Basic characteristics of the subjects showed linear increase in values of SBP, DBP and the serum concentrations of AST, ALT, FBG, GFR, hsCRP (p

Aim: To examine biochemical markers of adiponectin, total anti-oxidant status (TAOS) and high sensitvity C-reactive protein (hsCRP) in individuals with and without metabolic syndrome (MetS).Methods: A cross-sectional study on 36 non-MetS and 36 MetS subjects was undertaken in Jakarta. Measured indicators were adiponectin, TAOS and hsCRP, apart from weight, height, waist circumference (WC), systolic blood pressure (SBP), diastolic blood pressure (DBP), and fasting blood glucose (FBG). Odds ratio (OR) of adiponectin, TAOS and hsCRP were calculated to assess risk for the development of MetS. Median values were determined as cutoffs to define high and low values of each parameter. Relationships between adiponectin, TAOS and hsCRP with WC were analyzed by using Spearman correlation analysis, and the contributions of all indicators to the development of MetS were analyzed by using logistic regression.Results: Adiponectin dan hsCRP differed signifi cantly between non MetS and MetS subjects (4.2 + 1.4 vs 3.1 + 1.0 ug/mL) dan (0.97 + 0.92 vs 3.35 + 3.43 mg/L) (p < 0.01), but no signifi cant difference was found in TAOS (1.24 + 0.1 vs 1.28 + 0.2 mmol/L). Adiponectin associated negatively with WC (rs= -0.436; p < 0.01), while TAOS and hsCRP associated positively with WC (rs= 0.286, p = 0.02 and rs= 0.597, p < 0.01). The odds ratios (ORs) of adiponectin and hsCRP for the development of MetS were 4 (p = 0.01) and ~6,8 (p < 0.01), respectively; while the risk of subjects with adiponectinhs CRPratio of ≤ 2.31 to develop MetS was 25 times (p < 0.01) those with adiponectin-hsCRP ratio > 2.31.Conclusion: The use of adiponectin-hsCRP ratio increases the predictive power for the occurrence of MetS by 4-6 times the predictive power of adiponectin or hsCRP alone. (Med J Indones 2009; 18: 262-8)Keywords: adiponectin, hsCRP, total anti-oxidant status, metabolic syndrome.

BACKGROUND: Obesity is a major health problem in the world today. Obesity is closely associated with insulin resistance and type 2 diabetes. Epidemiological studies have shown that obese persons are in a state of insulin resistance, however, most of them do not progress to type 2 diabetes. This occurs because the beta cell function is still good enough for maintaining normal glucose level. Obestatin and visfatin are cytokines that are known to have a role in beta cell function. The aim of this study was to assess the relationship between visfatin and obestatin and Homeostasis Model Assessment of beta cell function (HOMA- β ) and Homeostasis Model Assessment of insulin resistance (HOMA-IR). METHODS: This was a cross-sectional study involving 80 central obesity men with waist circumference >90 cm, age 30-65 years old. Visfatin and obestatin were measured by ELISA method. Beta pancreas cell dysfunction and insulin resistance were calculated by HOMA model. RESULTS: Our study showed a correlation between visfatin and HOMA- β (r=0.244 and p = 0.029) and visfatin with HOMA-IR (r=0.287 and p=0.001) and no correlation was found between obestatin with HOMA- β (r=0.010 and p=0.990) and obestatin with HOMA-IR (r=0.080 and p=0.480). We also found visfatin and obestatin concentrations were fluctuative depending on the measurements of the waist circumferences. CONCLUSIONS: High visfatin and low obestatin concentration were independently associated with increased beta pancreas cell dysfunction and insulin resistance. KEYWORDS: obesity. visfatin, obestatin, beta cell dysfunction (HOMA- β ), insulin resistance (HOMA-IR)