人骨髓多能成体祖细胞的分离培养及向肝细胞样细胞分化的研究

Isolation, culture and inducing differentiation of human marrow multipotent adult progenitor cells into hepatocyte-like cells

目的 培养人骨髓多能成体祖细胞(multipotent adult progenitor cells,MAPCs)并研究其向肝细胞样细胞的分化。方法应用体外细胞培养技术将人骨髓中的单个核细胞从无血液疾病患者的髂嵴骨髓中分离出来后,用磁式细胞分离仪分离富集骨髓单个核细胞中的CD45^-HLA-DR^-细胞,把其接种于DMEM培养基中进行培养,到达对数生长期时,加入成纤维细胞生长因子-4(fibrob-last growth factor-4,FGF-4)并改用维持液来诱导其向肝细胞样细胞分化,最后用免疫细胞化学方法检测其分化情况。结果 成功地进行了人骨髓MAPCs的原代和传代培养,并在体外成功地把其诱导分化为具有肝细胞表型特征的细胞。结论 FGF-4可诱导人MAPCs向具有肝细胞表型特征的细胞发生分化。

Objective To cultivate multipotent adult progenitor cells (MAPCs)derived from human bone marrow and study their differentiation into hepatocyte-like cells. Methods Bone marrow mononuclear cells (BMMNCs) were collected from iliac.crest of patients who have no hematologic diseases. CD45- HLA-DR- cells were isolated and enriched from BMMNCs by magnetic cell sorter , then planted and cultivated in DMEM media. FGF-4 was added to maintain liquids to induce differentiation of CD45- HLA-DR- cells into hepatocyte-like cells when they achieved the logarithmic growth phase.The differentiation of MAPCs was tested by immunocytochemistry. Results Human MAPCs were successfully cultivated and differentiated into cells with phenotypic characteristics of hepatocyte in vitro. Conclusion FGF-4 can induce differentiation of human marrow MAPCs into cells with phenotypic characteristics of hepatocyte.