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ZHJ-multipotent adult progenitor cells differentiate into hepatocyte-like cells as co-cultured with human liver cell line L02
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- Author:
- No author available
- Journal Title:
- WORLD CHINESE JOURNAL OF DIGESTOLOGY
- Issue:
- 23
- DOI:
- 10.3969/j.issn.1009-3079.2006.23.007
- Key Word:
- 人骨髓来源多能成体祖细胞;肝细胞;直接共培养;间接共培养
Abstract： 目的:探索人骨髓来源多能成体祖细胞(ZHJ-MAPCs)在与人肝细胞系L02在体外共培养条件下诱导分化为肝细胞的可行性.方法:(1)间接共培养:将第4代ZHJ-MAPCs和人肝细胞系L02仅培养液相通行间接共培养.分别于培养第1、3、5、7天应用免疫细胞化学法鉴定ZHJ-MAPCs的ALB、AFP、CK-18、CK-19等肝细胞特征性表型表达情况,并设阳性对照和阴性对照,计数阳性细胞比率;(2)直接共培养:将CFSE荧光(绿色)标记的第4代ZHJ-MAPCs与L02(密度均为1×107/L)按照各50%比例混合行直接共培养.5 d后用SABC-Cy3免疫荧光(红色)双标后在激光共聚焦显微镜下分别观察ZHJ-MAPCs表达ALB,AFP,CK-18的情况.同样设阳性对照和阴性对照.结果:(1)间接共培养:AFP在ZHJ-MAPCs间接共培养第1天即表现为强阳性,随后表达逐渐减弱.ALB在共培养第1天有可疑阳性,第3天出现较强的阳性,第5天达到高峰.CK-18在培养第1、3天检测均为阴性,至第5天开始出现阳性,第7天表达较前增强.CK-19在各时间点均为阴性着色.(2)直接共培养:胞质内呈黄色荧光ALB和CK18的阳性细胞即向肝细胞分化的ZHJ-MAPCs在共培养5 d时出现较多,而AFP阳性表达则仅出现在极个别细胞,该结果与阳性对照细胞荧光表达类似.结论:人骨髓来源的ZHJ-MAPCs与人肝细胞系L02行间接或直接共培养均能够诱导其向成熟肝样细胞定向分化,且直接共培养有提前趋势.
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