富生长因子血小板:间充质干细胞培养中胎牛血清(FBS)的合适替代品

Fatemeh Hoseinpour Kasgari, Maryam Samareh Salavati Pour, A. Farsinejad, A. Fatemi, R. M. Khalilabadi
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引用次数: 1

摘要

简介:由于间充质干细胞具有很高的分化潜力和自我更新能力,目前研究人员广泛考虑将其用于多种疾病。FBS在培养基中用作MSC增殖、分化和其他培养过程的补充,这与各种感染的传播风险以及免疫反应有关。源自血小板的PRGF含有引起MSC生长和增殖的生长因子。本研究旨在比较PRGF与FBS对脐带来源的MSCs中h-TERT基因表达的影响。材料与方法:本研究为实验研究。从克尔曼输血中心获得四个过期的浓缩血小板袋,并通过血小板袋的多次离心制备PRGF。向PRGF中加入氯化钙作为抗凝剂,以防止培养基的凝胶化。另一方面,从脐带中分离间充质干细胞作为原代培养物。通过流式细胞术确认细胞的表型,并将细胞随机培养为对照组(使用FBS)和实验组(使用PRGF)。6天后,通过实时PCR技术研究了参与延长细胞寿命的基因(h-TERT)的表达。结果:成功地从脐带中分离出间充质干细胞。在形态学上,实验组(使用PRGF)培养的间充质细胞与对照培养基中的细胞相似。细胞表现出CD73、CD90和CD105的高表达水平,而造血细胞的表面标记物如CD45和CD34轻微表达。因此,对照组和实验组之间的细胞表面标记物表达没有显著差异。在本研究中,使用实时PCR技术,与对照相比,来自血小板的PRGF可以增加脐带间充质干细胞中h-TERT基因的表达。(P=0.034)
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Platelet Rich in Growth Factors (PRGF): A Suitable Replacement for Fetal Bovine Serum (FBS) in Mesenchymal Stem Cell Culture
 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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