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Effects of Serial Passage on the Characteristics and Cardiac and Neural Differentiation of Human Umbilical Cord Whartons Jelly-Derived Mesenchymal Stem Cells

机译:连续传代对人脐带沃顿胶质间充质干细胞的特性及其心脏和神经分化的影响

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摘要

Background and Objective. It is important to guarantee the quality of stem cells. Serial passage is the main approach to expand stem cells. This study evaluated effects of serial passage on the biological characteristics of human umbilical cord Wharton's jelly-derived MSCs (WJ MSCs). Methods. Biological properties of WJ MSCs in the early (less than 10 passages, P10), middle (P11–20), and late (more than P20) phases including cell proliferation, cell cycle, phenotype, senescence, oncogene expression, stemness marker expression, and differentiation capacity were evaluated using flow cytometry, real-time PCR, immunocytofluorescence, and western blot. Results. It was found that there were no significant differences in cell proliferation, cell cycle, phenotype, and stemness marker expression in different phases. However, the expression of senescence-related gene, p21, and oncogene, c-Myc, was significantly upregulated in the late phase, which had close relations with the obviously increased cell senescence. Moreover, cardiac differentiation capability of WJ MSCs decreased whereas the propensity for neural differentiation increased significantly in the middle phase. Conclusions. This study reveals that WJ MSCs in the early and middle phases are relatively stable, and effect of serial passage on the lineage-specific differentiation should be considered carefully.
机译:背景和目标。保证干细胞的质量很重要。连续传代是扩增干细胞的主要方法。这项研究评估了连续传代对人脐带沃顿氏胶来源的MSC(WJ MSC)生物学特性的影响。方法。 WJ MSC在早期(少于10代,P10),中期(P11–20)和晚期(大于P20)阶段的生物学特性,包括细胞增殖,细胞周期,表型,衰老,癌基因表达,干性标记物表达,使用流式细胞仪,实时荧光定量PCR,免疫细胞荧光和蛋白质印迹法评估其分化能力。结果。发现在不同阶段,细胞增殖,细胞周期,表型和干性标志物表达没有显着差异。然而,衰老相关基因p21和癌基因c-Myc的表达在晚期明显上调,这与细胞衰老明显增加密切相关。此外,在中期,WJ MSCs的心脏分化能力下降,而神经分化的倾向则明显增加。结论。这项研究表明,WJ MSC的早期和中期是相对稳定的,应仔细考虑系列传代对谱系特异性分化的影响。

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