大鼠骨髓间充质干细胞体外诱导分化为肝样细胞的研究

发布时间：2018-05-27 21:12

本文选题：骨髓间充质干细胞 + 肝细胞生长因子　；参考：《广西医科大学》2006年硕士论文

【摘要】： 目的:探讨体外诱导大鼠骨髓间充质干细胞(bone mesenchymal stem cells , BMSCs)定向分化为肝样细胞的可行性。方法:肝细胞生长因子(hepatocte growth factor ,HGF)体外诱导大鼠BMSCs,诱导第0d、7d、14d、21d,分别观察细胞形态特征;将培养出的细胞进行碱性磷酸酶染色;免疫细胞化学法检测甲胎蛋白(AFP)、细胞角质蛋白18(CK18)及白蛋白(ALB)的表达;糖原染色检测细胞糖原的表达;分别将基础培养基培养21d及用HGF定向诱导21d的细胞进行透射电镜观察其超微结构的变化。结果:HGF诱导组细胞从长梭形逐步转变为圆形,失去放射状排列的特点;非诱导组细胞呈长梭形生长,放射状或旋涡状排列。培养至第7d的细胞进行碱性磷酸酶染色呈阳性。HGF诱导组:第7d时AFP表达增加,但此后表达逐渐减少;CK18、ALB及糖原随诱导时间表达逐渐增强。在非诱导组:ALB、CK18、糖原在各时间点均呈阴性,AFP第0 d有少量表达,此后不再表达。定向诱导21d后的骨髓干细胞电镜下发现细胞呈肝索样排列,细胞间有大量的微绒毛,细胞内可见到丰富的粗面内质网、线粒体、溶酶体、吞饮泡,极类似于大鼠肝细胞电镜下的结构。结论:BMSCs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;其形态学上符合肝细胞的超微结构。
[Abstract]:Aim: to investigate the feasibility of inducing rat bone mesenchymal stem cells (BMSCs) to differentiate into hepatoid cells in vitro. Methods: hepatocyte growth factor (HGF) was used to induce rat BMSCs in vitro, and to observe the morphological characteristics of BMSCs on the 7th day, 14d and 21d, respectively, and the cultured cells were stained with alkaline phosphatase (ALP). Immunocytochemistry was used to detect the expression of alpha-fetoprotein (AFP), cytokeratin 18 (CK18) and albumin (ALB), and glycogen staining was used to detect the expression of glycogen. The ultrastructural changes of the cells cultured in basic culture medium for 21 days and induced by HGF for 21 days were observed by transmission electron microscope. Results the cells of the group induced by 1% HGF gradually changed from spindle shape to round shape and lost the characteristic of radial arrangement, while the cells in non-induced group grew in long spindle shape, radial or swirl shape. The expression of AFP increased on the 7th day, but the expression of CK18, ALB and glycogen increased gradually with the induction time. In the uninduced group, no expression of glycogen was found on day 0 of AFP at all time points, but no expression was found after that. Under electron microscope of bone marrow stem cells induced by directional induction for 21 days, the cells were found to be arranged like hepatic cord, with a large number of microvilli between the cells, and abundant rough endoplasmic reticulum, mitochondria, lysosomes, and swallowing vesicles were found in the cells. Very similar to the structure of rat hepatocytes under electron microscope. Conclusion HGF induced BMSCs differentiated into hepatocyte-like cells, and the expression of hepatocyte markers coincided with the development of hepatocytes, and its morphology was in accordance with the ultrastructure of hepatocytes.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329.2

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