Background: Due to their self-renewal and differentiation ability, the mesenchymal stem cells (MSCs) have been studied extensively. However, the MSCs lifespan is restricted; they undergo several divisions in vitro that cause several alternations in cellular features and relatively lessens their application. Thus, this study was aimed to assess the effect of platelet-derived microparticles (PMPs), a valuable source of proteins, microRNAs (miRNAs), and growth factors, on the expression of hTERT, c-MYC, p16, p53, and p21 as the most important aging and cell longevity genes alongside with population doubling time (PDT) of PMP-treated cells in comparison to a control group. Methods: Umbilical cord MSCs (UC-MSCs) were used in this study, whereby they reached a confluency of 30%. MSCs were treated by PMPs (50 µg/mL), and then, PDT was determined for both groups. Quantitative expression of hTERT, c-MYC, p16, p53, and p21 was examined through quantitative real-time PCR at various intervals (i.e. after five and thirty days as well as freezing-thawing process). Results: Our results demonstrated that the treated group had a shorter PDT in comparison to the control group (P<0.050). The real-Time PCR data also indicated that PMPs were able to remarkably up-regulate hTERT and c-MYC genes expression while down-regulating the expression of p16, p21, and p53 genes (P<0.050), especially following five days of treatment. Conclusion: According to these data, it appears that PMPs are a safe and effective candidate for prolonging the lifespan of UC-MSCs; however, further investigations are needed to corroborate this finding.
ntroduction: Due to high differentiation potential and self-renewality, Mesenchymal stem cells are now widely considered by researchers in several diseases. FBS is used as a supplement in culture media for proliferation, differentiation, and other culture processes of MSCs, which is associated with transmission risk of a variety of infections as well as immune responses. PRGF derived from platelets contains growth factors causing the growth and proliferation of MSCs. This study was conducted to compare the effect of PRGF in comparison to FBS on the expression of h-TERT gene, in umbilical cord-derived MSCs. Materials and Methods: This study is an experimental research. Four expired platelet concentrate bags were obtained from Kerman blood transfusion center, and PRGF was prepared by multiple centrifugation rounds of the platelet bag. Calcium chloride was added as an anticoagulant to PRGF in order to prevent gelatinization of the culture medium. On the other hand, mesenchymal stem cells were isolated from the umbilical cord as a primary culture. The phenotype of the cells was confirmed by flow cytometry, and the cells were randomly cultured as control (using FBS) and experimental (using PRGF) groups. The expression of the gene involved in increasing cell longevity (h-TERT) was investigated by real-time PCR technique after six days. Results: Mesenchymal stem cells were successfully isolated from the umbilical cord. Morphologically, the mesenchymal cells cultured in the experimental group (using PRGF) were similar to the cells in the control medium. The cells exhibited a high expression level of CD73, CD90, and CD105, while the surface markers of hematopoietic cells such as CD45 and CD34 were slightly expressed. Therefore, there was no significant difference in the expression of cell surface markers between control and experimental groups. In this study, using the real-time PCR technique, it was shown that PRGF derived from the platelet could increase the expression of h- TERT gene in the umbilical cord mesenchymal stem cells compared with the control. (P = 0.034)
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