体外不同诱导条件对大鼠胚胎肝干细胞分化影响的实验研究

发布时间：2018-06-28 15:18

本文选题：肝干细胞 + 肝细胞生长因子　；参考：《第四军医大学》2010年硕士论文

【摘要】： 目的探讨和比较不同体外条件对大鼠胚胎肝干细胞分化成熟的影响。方法将取自孕14天胎龄F344大鼠胚胎肝组织的肝干细胞分别接种至含10、20、40 ng/mL肝细胞生长因子(Hepatocyte Growth Factor HGF),0.5%、1%、2%二甲基亚砜(Dimethyl Sulfoxide DMSO),10 ng/mL HGF+0.5% DMSO、20 ng/mL HGF+1% DMSO、40 ng/mL HGF+2% DMSO以及空白的培养基中进行体外培养15 d;诱导15 d后,酶联免疫吸附实验(enzyme-linked immunosorbent assay ELISA)检测和比较不同组细胞内甲胎蛋白(alpha fetoprotein AFP)和白蛋白(albumin ALB)的浓度,实时定量PCR比较不同组细胞ALB、葡萄糖6-磷酸酶(Glucose-6-phosphatase G-6P)、细胞角蛋白8(Cytokeratin 8 CK-8)、细胞角蛋白18(Cytokeratin 18 CK-18)的mRNA的表达水平。结果诱导15 d后各实验组与空白对照组相比肝干细胞ELISA检测AFP明显下降(P0.01),ALB明显升高(P0.01);实时定量PCR结果表明各实验组ALB、G-6P、CK-8、CK-18的mRNA水平较对照组均明显升高,其中大鼠胎肝干细胞体外最佳诱导条件为20 ng/mL HGF+1% DMSO(P0.05)。结论 HGF和DMSO均可在体外诱导大鼠肝干细胞分化,不同浓度HGF和DMSO影响其分化成熟程度;HGF、DMSO对大鼠胎肝干细胞的分化具协同效应。
[Abstract]:Objective to investigate and compare the effects of different conditions on the differentiation and maturation of rat embryonic liver stem cells in vitro. Methods Hepatic stem cells derived from fetal liver tissue of F344 rats were inoculated into 10% 20 ~ 40 ng / mL Hepatocyte growth Factor (HGF) and 0.5% dimethyl Sulfoxide DMSO 10 ng / mL HGF 0.5% DMSO-1% DMSO-40 ng / mL HGF 2% HGF 2% HGF 20 ng / mL HGF 20 ng / mL HGF 1% DMSO-1% DMSO-40 ng / mL HGF 2%. DMSO and blank medium were cultured in vitro for 15 days, and after 15 days of induction, Enzyme linked immunosorbent assay (enzyme-linked immunosorbent assay) was used to detect and compare the concentrations of (alpha fetoprotein and albumin. The mRNA levels of ALB, Glucose-6-phosphatase G-6P, cytokeratin 8 CK-8 and cytokeratin 18 CK-18 were compared by real-time quantitative PCR. Results after 15 days of induction, the levels of AFP in liver stem cell Elisa were significantly decreased (P0.01) and ALB was significantly increased (P0.01) in each experimental group compared with the control group, and the mRNA level of CK-8 CK-18 in all experimental groups was significantly higher than that in the control group. The optimal induction condition of rat fetal liver stem cells in vitro was 20 ng / mL HGF 1% DMSO (P0.05). Conclusion both HGF and DMSO can induce the differentiation of rat hepatic stem cells in vitro. Different concentrations of HGF and DMSO have synergistic effects on the differentiation of rat fetal liver stem cells.
【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R329

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