诱导大鼠骨髓Thy-1^+ β_2M^-细胞分化为肝脏细胞  

Differentiation of rat bone marrow derived Thy-1^+ β_2M^-cells into hepatocytes

作　　者：王韫芳[1] 南雪[1] 李艳华[1] 张锐[1] 闫舫[1] 岳文[1] 裴雪涛[1] 

机构地区：[1]军事医学科学院输血医学研究所干细胞生物学研究室,北京100850

出　　处：《中华血液学杂志》2005年第2期69-73,共5页Chinese Journal of Hematology

基　　金：国家高技术"863"计划领域重大专项基金资助项目(2002AA205051和2003AA205160);国家重点基础研究发展规划(937)资助项目(2001CB509906);北京市科委课题资助项目(H020220010190)

摘　　要：目的探讨定向诱导大鼠骨髓来源的Thy1+β2M-细胞(bonemarrowderivedThy1+β2M-cell,BDTC)分化为成熟的、有功能的肝脏细胞及其可能的机制。方法分别采用Transwell培养板联合培养BDTC和丙烯醇(allylalcohol,AA)损伤的肝脏细胞,并用含有肝细胞生长因子和碱性成纤维细胞生长因子的条件诱导液单独培养BDTC,通过光镜和电镜观察BDTC的形态变化,免疫细胞化学和RTPCR检测BDTC诱导前后肝细胞特异性基因的表达;用靛青绿(indocyaninegreen,ICG)摄取、排泌实验及白蛋白、尿素分泌检测肝细胞的相关功能。结果联合培养及单独以条件诱导液培养7d,部分BDTC体积明显增大,出现大而圆的单个或多个细胞核,胞浆富含线粒体、内质网和糖原颗粒等;这些细胞表达幼稚或成熟肝细胞特异性的AFP、OV6、CK18等基因,并具有靛青绿摄取、排泌和分泌白蛋白、氨基代谢生成尿素等功能。结论大鼠BDTC在AA所致肝损伤环境中或在特定诱导体系内均可以向成熟的、有生理功能的肝脏细胞分化,而且该过程无需通过细胞融合而实现。Objective To investigate the differentiation of bone marrow derived Thy-1^+ β_~2 M^- cells (BDTC) into mature and functional liver cells and its mechanism. Methods BDTC were cocultured with allyl alcohol (AA)-injured hepatocytes and cultured alone in conditional medium containing HGF and bFGF, respectively. BDTC morphologic transformation was observed with phase-contrast and electron-microscopy. Hepatocyte-specific gene expression in cultured BDTC was identified by immunocytochemistry and reverse transcription-polymerase chain reaction (RT-PCR). Indocyanine green(ICG) ingestion/excretion and urea, albumin production were carried out to evaluate hepatocyte-related function. Results Some BDTC derived hepatocyte-like cells with high nuclear to cytoplasmic ratio containing mono-or multi-nuclei and abundant mitochondria, endoplasmic reticulum and glycogenic granules appeared after 7-day culture in both the two culture systems. These cells expressed hepatocyte-specific genes (AFP, OV-6, CK18, etc.), and possessed functions of ICG uptake, albumin production and ammonium metabolism. Conclusion Rat BDTCs could differentiate into mature and functional liver cells in special stimulation systems. Moreover, these differentiations were realized by “transdifferentiation”, and might dispense with “cell fusion”.

关 键 词：肝脏细胞 诱导 大鼠 Β2M 骨髓 白蛋白 肝细胞 分泌 细胞融合 细胞核 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]