摘要
目的探索小鼠骨髓来源的单个核细胞向肝细胞分化的诱导条件。方法通过密度梯度离心的方法获得小鼠骨髓来源的单个核细胞;在含有100g/L新生牛血清的IMDM培养液中添加细胞因子HGF20ng/ml、FGF-4 10ng/ml和OSM10ng/ml,诱导骨髓来源的单个核细胞向肝细胞分化。结果小鼠骨髓单个核细胞在3种细胞因子的诱导下,随着时间的延长,体积逐渐增大、形态上向上皮样细胞转变,胞浆丰富,出现双核或多核细胞;半定量RT-PCR的结果显示,甲胎蛋白(AFP)和细胞角蛋白19(CK19)的表达量在诱导初期首先增加,在诱导后期逐渐减少,而白蛋白(ALB)、细胞角蛋白18(CK18)以及酪胺酸胺基转移酶(TAT)的表达与诱导时间呈线性关系;免疫荧光检测诱导3周后的细胞,AFP、CK19、CK18和ALB均为阳性表达;诱导3周的细胞PAS糖原合成反应呈阳性,说明已经具备糖元合成和储存这一肝细胞的特征性功能。结论培养液中添加3种细胞因子(HGF、FGF-4和OSM)组合可以诱导小鼠单个核细胞向肝细胞分化。
Objective To establish an effective method for inducing mouse bone marrow mononuclear cells （MNCs） to differentiate into hepatocytes. Methods MNCs were isolated by gradient density eentrifugation, and the cells were cultured in Iseove＇s Modified Dulbecco＇s medium supplemented with 10% new bovine serum （NBS）, 20 ng/ml hepatocyte growth factor （HGF）, 10 ng/ml fibroblast growth factor-4 （FGF-4） and 10 ng/ml oncostatin m （OSM） for 21 days′ induction. The medium was changed every 3 days. Results When MNCs were cultured with HGF, FGF-4 and OSM, cuboidal morphology was observed, and cells also expressed marker genes specific for liver cells in a time-dependent manner. α-fetoprotein （AFP） and cytokeratin 19 （CK19） were expressed on the day 7, and CK18 and TAT were detected on the day 14-21, in common with the immunofluoresence results. Differentiated cells further demonstrated these cells also acquired functional characteristics of hepatocytes. Conclusion Mouse MNCs can differentiate into hepatocytes when induced by HGF, FGF-4 and OSM.
出处
《中华器官移植杂志》
CAS
CSCD
北大核心
2006年第6期330-333,共4页
Chinese Journal of Organ Transplantation
基金
绍兴市科技计划项目(2005141)
关键词
抗原
分化
髓单核细胞
肝细胞
Antigens
differentiation
myelomonocytic
Hepatocytes
