دوشنبه 26 مرداد 1405
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Ethics code: IR.TUMS.SPH.REC.1397.319
چکیده: (17 مشاهده)
Background and objective: Exosomes derived from umbilical cord blood mesenchymal stem cells (UCB-MSCs) may enhance hematopoietic stem cells (HSCs) proliferation while preserving their undifferentiated state. This study evaluates the potential of UCB-MSC-derived exosomes as a cell-free strategy to expand HSCs, addressing the limitation of low HSC numbers for hematological disorder therapies.
Methods: In this in vitro study, MSC-derived exosomes (MSC-EXOs) were isolated and characterized. Subsequently, three umbilical cord blood samples were collected, and CD34+ HSCs were purified using magnetic-activated cell sorting (MACS). To identify the optimal exosome concentration, CD34+ cells were cultured with MSC-EXOs at four different concentrations (1, 5, 10, and 100 ng/mL). Gene expression levels of CXCR4, VLA-4, VLA-5, hTERT, and HOXA9 were assessed using quantitative RT-PCR, while colony-forming cell (CFC) assays were performed to evaluate the clonogenic potential of HSCs.
Results: After 7 days of co-culture, an increase in mononuclear cells (MNCs) was observed across all groups. Notably, the expression of the CD34 marker was significantly highest in Group III (71%) and Group IV (approximately 48%), exceeding levels seen in the other groups. Gene expression analysis revealed a significant upregulation of target genes in Group IV. Consistent with these findings, the CFC assay showed that Group IV had a significantly higher colonization rate than the other experimental conditions.
Conclusion: The results demonstrate that MSC-derived exosomes can effectively mimic the supportive functions of their parent cells in vitro, thereby significantly enhancing HSC proliferation.
Methods: In this in vitro study, MSC-derived exosomes (MSC-EXOs) were isolated and characterized. Subsequently, three umbilical cord blood samples were collected, and CD34+ HSCs were purified using magnetic-activated cell sorting (MACS). To identify the optimal exosome concentration, CD34+ cells were cultured with MSC-EXOs at four different concentrations (1, 5, 10, and 100 ng/mL). Gene expression levels of CXCR4, VLA-4, VLA-5, hTERT, and HOXA9 were assessed using quantitative RT-PCR, while colony-forming cell (CFC) assays were performed to evaluate the clonogenic potential of HSCs.
Results: After 7 days of co-culture, an increase in mononuclear cells (MNCs) was observed across all groups. Notably, the expression of the CD34 marker was significantly highest in Group III (71%) and Group IV (approximately 48%), exceeding levels seen in the other groups. Gene expression analysis revealed a significant upregulation of target genes in Group IV. Consistent with these findings, the CFC assay showed that Group IV had a significantly higher colonization rate than the other experimental conditions.
Conclusion: The results demonstrate that MSC-derived exosomes can effectively mimic the supportive functions of their parent cells in vitro, thereby significantly enhancing HSC proliferation.
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