摘要
目的:探讨肝损伤微环境在脐血来源的细胞转化为类肝细胞中的作用。方法:采用双室培养法将分离后的人脐血单个核细胞(mononuclearcells,MNC)与小鼠损伤肝脏共培养;分别采用细胞化学染色、反转录-聚合酶链反应(reversetranscriptionpolymerasechainreaction,RT-PCR)和基因序列分析方法对培养的细胞进行检测,以获取人脐血MNC向类肝细胞转化的相关信息。结果:在与小鼠损伤肝脏共培养72h的人脐血MNC中发现大量过碘酸-Schiff染色阳性细胞;RT-PCR显示,与小鼠损伤肝脏共培养48h和72h后的人脐血MNC中含有肝细胞标志物人清蛋白(humanalbumin,hALB)和人哺乳类动物转录因子-4(hGATA-4)mRNA表达阳性细胞,并通过序列分析确认所得到的hALB和hGATA-4目标片段核苷酸序列与Gene-Bank中的参考序列相吻合,而人脐血MNC与正常小鼠肝脏共培养组、单独培养的人脐血MNC组以及小鼠肝脏细胞的同类对比试验均呈阴性结果。结论:在模拟的肝损伤微环境中,人脐血来源的细胞具有向类肝细胞转化的趋向。
Objective ： To explore the effect of the microenvironment induced by damaged mouse hepatic cells on the conversion of human umbilical cord blood-derived cells into hepatocyte-like cells. Methods： A hepatic injury-like microenvironment was mimicked using carbon tetrachloride damaged mouse hepatic cells, where mononuclear cells （MNC） from human umbilical cord blood were cultured in a compartment separated by trans-well membrane. Histochemical staining, reversed transcription-polymerase chain reaction （RT-PCR） and gene sequencing were performed for the information on the conversion of human umbilical cord blood MNC. Results： A number of PAS positive stained cells in MNC co-cultured with damaged mouse hepatic cells were observed after 72 h. Cells expressing mature hepatocyte markers, human albumin （hALB） and human GATA-4 （hGATA-4） mRNA were detected by RT-PCR, which was further confirmed with sequencing. Relevant control groups, MNC co-cultured with normal mouse hepatic cells and MNC cultured alone remained negative. Conclusion： The culture system using damaged mouse hepatic cells as stimulator could be a potential in vitro system for the conversion of human umbilical cord blood-derived cells into henatocyte-like cells.
出处
《北京大学学报（医学版）》
CAS
CSCD
北大核心
2005年第4期402-405,共4页
Journal of Peking University:Health Sciences
基金
深圳市科技计划项目(200204111)~~
关键词
胎血
单核细胞
肝损伤
肝细胞
共同培养
Fetal blood
Monocytes
Hepatic injury
Hepatocytes
Coculture
作者简介
Corresponding author＇ s e-mail, zftft@ yahoo. com. cn
