胎肝源性细胞促进人骨髓间充质干细胞向肝细胞的定向分化  

Fetal liver-derived cells stimulate differentiation of human mesenchymal stem cells into hepatocytes in vitro

作　　者：汪泱[1] 车媛梅[2] 张一[2] 谢安[1] 崔苏萍[1] 娄远蕾[1] 

机构地区：[1]南昌大学第一附属医院泌尿外科研究所,南昌330006 [2]南昌大学第一附属医院感染科,南昌330006

出　　处：《解剖学杂志》2009年第5期599-602,共4页Chinese Journal of Anatomy

基　　金：江西省科技厅重点项目;江西省自然科学基金（2007GZY1470）

摘　　要：目的：探讨人胎肝源性细胞（HFLC）对骨髓间充质干细胞（MSCs）体外诱导分化为肝系细胞的效应和机制。方法：药物流产的胎肝源性细胞和Hoechst33342标记的人MSCs，按培养方式不同，分为HFLC和MSCs直接接触共培养组及transwell共培养组，用倒置显微镜观察细胞形态，在不同时段用免疫细胞荧光法检测肝系细胞标志如AFP、CK-18和CK-19并进行糖原染色。结果：在直接接触共培养组，免疫细胞荧光法显示分化细胞表达AFP、CK-18和CK-19，并随分化时间延长表达量增加；同时，分化细胞具有贮存糖原功能，PAS染色显示分化细胞糖原染色阳性。而在transwell共培养组，分化细胞未检测到肝系细胞标志物的表达及糖原的贮存。结论：HFLC与MSCs的直接接触共培养体系的微环境，有利于MSCs定向分化为肝系细胞。Objective： To investigate effects of human fetal liver-derived cell （HFLC） on differentiation of human mesenchyreal stem cells （MSCs） into hepatocytes. Methods： Fetal hepatocytes from medical abortion and adult human MSCs labeled with Hoechst 33342 were co-cultured directly and/or separately in a dual chamber dish. Cell morphology was observed under a phase microscope. Hepatocyte specific markers alpha-fetoprotein （AFP）, cytokeratin （CK）18 and CK19 were identified by immunofluoresent staining and glycogen storage was detected by Periodic Acid-Sehiff （PAS） staining at different time intervals. Results： In the direct co-culture system, immunofluorescent staining demonstrated that the differentiated cells had the ability of expressing AFP, CK18 and CK19, and increased with time of differentiation. In addition, the differentiated cells had functional properties of hepatoeytes, such as glycogen storage, which was evaluated by PAS staining. This phenomenon did not appear when HFLC were separated from human MSCs in a dual chamber dish. Conclusion： The direct co-culture of HFLC with adult human MSCs may create a particularly favorable microenvironment for the transdifferentiation of the latter into hepatocyte-like cell, which may serve as a cell source for cell therapy of hepatic tissues.

关 键 词：骨髓间充质干细胞 胎肝源性细胞 共培养 肝细胞 分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R735.7[医药卫生—基础医学]