Investigation the Effect of Serum Rich in Growth Factors on Proliferation, Growth and Expression of Genes Involved in Cell Longevity by Mesenchymal Stem Cells

R. M. Khalilabadi, Fatemeh Hoseinpour Kasgari
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Abstract

Background: The limited lifespan of Mesenchymal Stem Cells (MSCs) has highly restricted their application. The Serum Rich in Growth Factors (SRGF) contains growth factors that are involved in processes of the growth and proliferation of MSCs. The present study was aimed to examine the regulatory effects of SRGF on the expression of some genes which effect both proliferation and lifespan of MSCs. Methods: SRGF was obtained from platelets and MSCs were isolated from umbilical cord. The MSCs morphology and phenotype have been analyzed using phase-contrast microscope and flow cytometry, respectively. Cells were cultured either in presence of FBS 10% (as control) or SRGF 5% plus FBS 5% and FBS 10% alone (as tests). The cell Population Doubling Time (PDT) was measured hourly. The expression of related genes was analyzed employing real-time PCR technique. Findings: Finding of the present study showed that the experimental groups were morphologically and phenotypically as similar as to control group. We observed that the PDT in the experimental group was shorter than that was found in the control. We have also found that the expression of hTERT and c-MYC genes was increased, while, and P16 and P53 genes were down- regulated. These results were superior in the 10%SRGF group than in the 5% SRGF + 5% FBS group. Conclusion: According to the finding of this study, SRGF could possibly serve as an effective proliferative and lifespan inducing factor for MSCs. These results also indicated that SRGF has the tendency to be employed as an appropriated alternative for FBS in cell culture.
富生长因子血清对间充质干细胞增殖、生长及细胞寿命相关基因表达影响的研究
背景:间充质干细胞(Mesenchymal Stem Cells, MSCs)有限的寿命严重限制了其应用。富生长因子血清(SRGF)含有参与骨髓间充质干细胞生长和增殖过程的生长因子。本研究旨在探讨SRGF对影响间充质干细胞增殖和寿命的基因表达的调控作用。方法:从血小板中获得SRGF,从脐带中分离MSCs。利用相差显微镜和流式细胞术分别对MSCs的形态和表型进行分析。细胞分别在10%的FBS(作为对照)或5%的SRGF + 5%的FBS和单独10%的FBS(作为试验)中培养。每小时测量细胞群倍增时间(PDT)。采用实时荧光定量PCR技术分析相关基因的表达。结果:本研究发现,实验组与对照组在形态和表型上相似。我们观察到实验组的PDT比对照组短。我们还发现hTERT和c-MYC基因表达增加,P16和P53基因表达下调。这些结果在10%SRGF组优于5% SRGF + 5% FBS组。结论:根据本研究发现,SRGF可能是一种有效的MSCs增殖和寿命诱导因子。这些结果也表明,SRGF在细胞培养中有被用作FBS的合适替代品的趋势。
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