损伤肝脏条件培养液体外诱导小鼠骨髓间充质干细胞分化为肝细胞的研究

发布时间：2018-08-09 13:16

【摘要】： 成体干细胞跨越分化为其它组织类型细胞的研究已成为当前干细胞研究的新热点，，它不仅对探索细胞分化发育的机制具有重要的理论意义，而且为临床细胞移植治疗开辟了新的途径。作为最重要的成体干细胞之一，骨髓来源的干细胞分化研究倍受重视，以往的研究表明，它们具有较强的可塑性，能够分化成神经细胞、心肌细胞、骨骼肌细胞、上皮细胞和胰岛细胞。近年来，Petersen等进一步发现，在CCl_4致肝脏损伤的微环境下，移植的骨髓细胞能够分化成为肝卵圆细胞甚至成为成熟的肝细胞和胆管细胞。然而，由于体内研究背景的复杂性，目前这种跨越分化的机制尚不清楚，这就需要建立相关的体外分化技术进行研究。另外，骨髓中存在能分化为肝细胞的干细胞虽已成定论，但究竟是其中的几个亚型还是所有的骨髓细胞均可分化为肝细胞仍值得研究。本文将注射过CCl_4的小鼠肝脏进行培养，收集损伤肝脏条件培养液，添加到培养液中以模拟体内肝损伤微环境，诱导间充质干细胞的分化。利用逆转录聚合酶链反应(RT-PCR)和免疫荧光反应检测各组细胞中甲胎蛋白(AFP)、细胞角蛋白18(CK18)、酪胺酸胺基转移酶(TAT)和白蛋白(ALB)基因的表达，并利用高碘酸希夫反应检测细胞糖原合成。结果显示损伤肝脏条件培养液可以成功诱导骨髓间充质干细胞体外分化成肝细胞。从而为骨髓间充质干细胞跨越分化为肝细胞的机制研究打下基础，同时也为体外利用骨髓细胞分化为肝细胞的相关临床应用研究，建立新的技术方法。
[Abstract]:The study of adult stem cells crossing into other types of cells has become a new focus of stem cell research. It is not only of great theoretical significance to explore the mechanism of cell differentiation and development. It also opens up a new way for clinical cell transplantation therapy. As one of the most important adult stem cells, stem cells derived from bone marrow have attracted much attention. Previous studies have shown that they can differentiate into nerve cells, cardiomyocytes and skeletal muscle cells. Epithelial cells and islet cells. In recent years, Petersen et al further found that in the microenvironment of liver injury induced by CCl_4, transplanted bone marrow cells can differentiate into hepatic oval cells and even become mature hepatocytes and bile duct cells. However, due to the complexity of in vivo research background, the mechanism of in vitro differentiation is not clear, which requires the establishment of in vitro differentiation technology. In addition, although the existence of stem cells that can differentiate into hepatocytes in bone marrow has been decided, it is still worth studying whether some of these subtypes or all bone marrow cells can differentiate into hepatocytes. In this paper, the liver of mice injected with CCl_4 was cultured, and the conditioned medium of liver injury was collected and added to the medium to simulate the microenvironment of liver injury in vivo and induce the differentiation of mesenchymal stem cells. Reverse transcriptase polymerase chain reaction (RT-PCR) and immunofluorescence reaction were used to detect the expression of keratin 18 (CK18), tyramidine aminotransferase (TAT) and albumin (ALB) gene in (AFP), cells. The results showed that the liver injury conditioned medium could induce the differentiation of bone marrow mesenchymal stem cells into hepatocytes in vitro. It will lay a foundation for the study of the mechanism of bone marrow mesenchymal stem cells to differentiate into hepatocytes, at the same time, it can also be used to study the clinical application of bone marrow mesenchymal stem cells differentiation into hepatocytes in vitro, and establish a new technique and method.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329

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