体外诱导大鼠骨髓干细胞分化为肝样细胞的实验研究

发布时间：2018-03-27 12:31

本文选题：骨髓干细胞　切入点：肝细胞生长因子　出处：《广西医科大学》2005年硕士论文

【摘要】：目的:探讨大鼠骨髓干细胞在肝细胞生长因子(HGF)诱导下分化为肝样细胞的可能性及分化特性。方法:HGF体外诱导大鼠骨髓干细胞向肝样细胞分化,诱导第O、7、14、21天,观察细胞形态特征;免疫细胞化学法检测甲胎蛋白(AFP)、细胞角质蛋白18(CKl8)的表达;PAS检测细胞糖原;逆转录聚合酶链反应(RT-PCR)法检测AFP mRNA、ALB mRNA水平的表达。结果:①新鲜分离的骨髓干细胞AFP mRNA呈弱阳性,ALB mRNA阴性。在非诱导组:AFPmRNA、ALB mRNA、AFP、CK18、糖原在各时间点均呈阴性。HGF诱导组:7天时AFP mRNA、AFP表达明显增加,但此后表达逐渐减少ALB mRNA、CK18、糖原随诱导时间表达逐渐增强。②HE染色较深的小圆形细胞在非诱导条件下其直径未发生明显变化;诱导组HE染色较深的小圆形细胞与AFP、CK18、糖原染色阳性的细胞形态大小一致,随诱导时间延长其直径增大。结论:①大鼠骨髓干细胞在HGF诱导下分化为肝样细胞,肝细胞标志物表达与肝细胞发育过程吻合。②骨髓源性肝干细胞的形态学特征为集落中HE染色较深的小圆形细胞。
[Abstract]:Objective: to investigate the possibility and characteristics of rat bone marrow stem cells differentiating into hepatoid cells induced by hepatocyte growth factor (HGF). Methods: rat bone marrow stem cells were induced to differentiate into hepatocyte-like cells in vitro, and OF71421 days were induced. The morphological characteristics of the cells were observed, the expression of AFP and CKl8 were detected by immunocytochemistry and the glycogen was detected by pas. Reverse transcriptase polymerase chain reaction reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the expression of AFP mRNA-ALB mRNA in bone marrow stem cells isolated from fresh bone marrow stem cells. Results AFP mRNA of bone marrow stem cells isolated from freshly isolated bone marrow stem cells was weakly positive for ALB mRNA negative. In the non-induced group, AFP mRNA-ALB mRNA-AFPmRNA-AFPmRNA-AFPCK18, glycogen was negative at all time points .HGF-induced AFP mRNA was negative for 7 days. The expression of AFP mRNA-AFP was significantly increased. After that, the expression of ALB mRNA-CK18 was gradually decreased, and the diameter of the small circular cells stained deeply with the induction time was not changed significantly. In the induction group, the size of the small round cells stained deeply by HE was the same as that of AFP-CK-18, and the size of the cells with positive glycogen staining was the same, and the diameter of the cells increased with the prolongation of the induction time. Conclusion: the bone marrow stem cells of rats with HGF can differentiate into hepatoid cells. The expression of hepatocyte markers coincides with the development of hepatocytes. 2 the morphological characteristics of bone marrow-derived hepatic stem cells are small round cells stained deeply by HE in the colony.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R329.2

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