One of the main causes of acute respiratory distress syndrome in coronavirus disease 2019 (COVID-19) is cytokine storm, although the exact cause is still unknown. Umbilical cord mesenchymal stromal cells (UC-MSCs) influence proinflammatory T-helper 2 (Th 2 ) cells to shift to an anti-inflammatory agent. To investigate efficacy of UC-MSC administration as adjuvant therapy in critically ill patients with COVID-19, we conducted a double-blind, multicentered, randomized controlled trial at four COVID-19 referral hospitals in Jakarta, Indonesia. This study included 40 randomly allocated critically ill patients with COVID-19; 20 patients received an intravenous infusion of 1 Â 10 6 /kg body weight UC-MSCs in 100 ml saline (0.9%) solution (SS) and 20 patients received 100 ml 0.9% SS as the control group. All patients received standard therapy. The primary outcome was measured by survival rate and/or length of ventilator usage. The secondary outcome was measured by clinical and laboratory improvement, with serious adverse events. Our study showed the survival rate in the UC-MSCs group was 2.5 times higher than that in the control group (P = .047), which is 10 patients and 4 patients in the UC-MSCs and control groups, respectively. In patients with comorbidities, UC-MSC administration increased the survival rate by 4.5 times compared with controls. The length of stay in the intensive care unit and ventilator usage were not statistically significant, and no adverse events were reported. The application of infusion UC-MSCs significantly decreased interleukin 6 in the recovered patients (P = .023). Therefore, application of intravenous UC-MSCs as adjuvant treatment for critically ill patients with COVID-19 increases the survival rate by modulating the immune system toward an antiinflammatory state.
: Human adipose derived stem cells (hADSCs) can be cultured in outdated platelet lysate (PL) containing medium. Processing of thrombocyte concentrate to get PL can be done by various freeze-thaw cycles. There was no information whether various processed PL containing media gave the same proliferation potential and surface marker expressions. Therefore, the aim of this study was to know the proliferation and surface marker characteristics of hADSCs after expansion in various processed PL containing media. hADCSs were cultured in various processed PL containing media, namely F1, F2, and F3, where the numbers denoted once, twice and three times freeze-thaw cycles. Proliferations were measured on day-2, day-4, day-7, and day-10. Positive surface markers were CD90, CD73, and CD105, while negative markers were a cocktail of CD34, CD45, CD11b, CD19, and HLA-DR. Difference in proliferations and surface marker expressions between F1, F2, and F3 were analyzed by one-way ANOVA. We found that cell proliferation in F1, F2, and F3 showed no significant difference on day-7 and day-10. The expression levels of CD90 and CD 105 in F1, F2, and F3 were all above 90%, which correspond with mesenchymal stem cells according to the requirement of International Society for Cell Therapy (ISCT). However, CD73 showed highest (97%) expression on F2 and lowest on F3 (81.8%). Negative marker range was 2.4-2.9%. In Conclusion, at the end of culture, hADSCs showed similar profile and proliferation, when cultured in F1, F2, and F3.
A liver organoid is an in vitro reconstruction of the liver that mimics the in vivo liver microstructure and performs liver functions. Liver organoids can be used for drug testing, as a model of liver disease pathogenesis, and as a bioartificial liver prototype material to develop promising alternative therapies for liver failure. In this study, we reconstructed liver organoids using primary rat hepatocytes, a hepatic stellate cell line (LX2), human umbilical cord-mesenchymal stem cells (UC-MSCs), and human umbilical cord blood (UCB)-CD34+ hematopoietic stem/progenitor cells. Suspensions of primary rat hepatocytes, LX2 cells, UC-MSCs, and UCB-CD34+ cells were co-cultured using 11 ratio sets, and spheroid formation was evaluated for 2 days. Ratio sets with a positive liver organoid appearance were cultured in four different culture media, and after they were harvested, their viability was compared with that of a hepatocyte monoculture. Liver organoids were further analyzed for 14 days to assess albumin and urea production as well as relative gene expression. We found that a 5:1:2:2 cellular density ratio of hepatocytes:LX2 cells:UC-MSCs:UCB-CD34+ cells, respectively, and Williams E medium supplemented with platelet lysate, ITS, and dexamethasone were the most suitable conditions for liver organoid reconstruction. Expression of the albumin and GPT1 genes and CD31 in the liver organoid increased until day 14, while urea secretion increased until day 5. Liver organoids reconstructed through the 3D co-culture of primary rat hepatocytes, LX2 cells, UC-MSCs, and UCB-CD34+ cells at a specific cellular ratio using an economical medium with a simple composition maintained their functions until day 14. As a future direction, these organoids can be used to develop a bioartificial liver.
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