Cx43基因干扰对鼠胎肝干细胞培养的优化效应

发布时间：2018-06-05 02:42

本文选题：肝细胞 + 连接蛋白　；参考：《中国组织工程研究》2015年14期

【摘要】：背景:胎肝干细胞具有分化成肝细胞、胆管细胞的潜能,参与肝脏的修复与重建,是肝细胞的重要来源,但是人体的胎肝干细胞含量极少,如何获取一定数量和较高纯度的胎肝干细胞是目前研究的热点。目的:构建能有效抑制大鼠胎肝干细胞Cx43基因表达的siR NA载体,探讨抑制Cx43表达对体外培养的鼠胎肝干细胞增殖及细胞周期的影响。方法:体外培养鼠胎肝干细胞,设计及合成靶向Cx43的siR NA序列(Cx43-siR NA)以及阴性对照序列(NC-si RNA),采用电转法转染大鼠胎肝干细胞,即为实验组和对照组,未转染的胎肝干细胞为空白组。应用real-time PCR和Western blot法检测转染前后鼠胎肝干细胞Cx43基因和蛋白的表达;细胞生长曲线、CCK-8法观察细胞生长增殖情况;流式细胞术测定细胞周期分布的变化。结果与结论:转染Cx43-siR NA后,实验组与对照组、空白组相比Cx43基因和蛋白水平表达均明显降低,细胞的生长速度明显增快,G0/G1期细胞减少,S期细胞数增多,差异有显著性意义(P0.05),结果表明通过电转法转染靶向Cx43的si RNA能促进体外培养的鼠胎肝干细胞增殖,对其培养有优化作用。
[Abstract]:Background: fetal liver stem cells (FHSCs) have the potential to differentiate into hepatocytes and bile duct cells and participate in the repair and reconstruction of liver, which is an important source of hepatocytes. How to obtain a certain number and high purity of fetal liver stem cells is a hot topic at present. Aim: to construct a siR na vector that can effectively inhibit the expression of Cx43 gene in rat fetal liver stem cells (FHSCs), and to investigate the effect of inhibiting Cx43 expression on the proliferation and cell cycle of rat fetal liver stem cells (FHSCs) cultured in vitro. Methods: rat fetal liver stem cells were cultured in vitro. The siR na sequence (Cx43-siR NAA) targeting Cx43 and the negative control sequence (NC-si RNAs) were designed and synthesized. Rat fetal liver stem cells were transfected by electrotransposed method, that is, the experimental group and the control group. The untransfected fetal liver stem cells were blank group. Real-time PCR and Western blot methods were used to detect the expression of Cx43 gene and protein in fetal liver stem cells before and after transfection, cell growth curve CCK-8 method was used to observe cell growth and proliferation, and cell cycle distribution was measured by flow cytometry. Results and conclusion: after transfection of Cx43-siR na, the expression of Cx43 gene and protein in the experimental group and the control group were significantly lower than those in the control group, and the cell growth rate was significantly increased. The results showed that si RNA targeting Cx43 could promote the proliferation of rat fetal liver stem cells in vitro and optimize its culture.
【作者单位】：天津市第四中心医院肝胆外科;天津市南开医院微创外科;
【分类号】：R575

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