摘要
目的：利用肝细胞生长因子（HGF）、表皮生长因子（EGF）和成纤维细胞生长因子-4（FGF-4）诱导骨髓间充质干细胞（MSCs）向肝细胞分化,为肝组织工程提供新的细胞来源。方法：取志愿者骨髓,使用密度梯度离心法联合贴壁筛选法获取MSCs,用含有HGF、EGF、FGF的培养基进行培养。用流式细胞仪、免疫细胞化学法鉴定细胞表型,检测诱导后细胞胆红素处理能力。结果：骨髓MSCs生长良好,表型为CD29阳性,CD34阴性。诱导后细胞可在21～28 d时形成肝样细胞。诱导7 d后细胞检测到AFP的表达,第14天时CK18呈阳性表达,随着诱导时间的延长,阳性率增加,并能显著降低胆红素浓度。结论：HGF、EGF、FGF联合能诱导骨髓MSCs分化为具有处理胆红素能力的肝样细胞。
Objective To explore the possibility of differentiation of mesenchymal stem cells （MSCs） into hepatocyte-like cells induced by hepatocyte growth factor（HGF）, epidermal growth factor（EGF） and fibroblast growth factor （FGF）, and to find a new cell source for liver tissue engineering. Methods Bone marrow cells were obtained from volunteers, hMSCs were separated by density gradient centrifugation and were cultured through adhere culture. MSCs were cultured in DMEM/F12 medium with HGF, EGF, FGF. The phenotypes of cells were identified by flow cytometry, immunohistochemistry, at last the hepatocyte-like cells were cultured in DMEM/F12 medium with bilirubin. Results The MSCs, collected were CD29 positive and CD34 negative, the MSCs induced became hepatocyte-like cells and immunocytochemieal analysis for CK18 and AFP showed positive staining reaction. The serum concentration of TBIL in treatment group was significantly decreased （P 〈 0.05 ）. Conclusion HGF, EGF and FGF can induce MSCs into hepatocyte-like cells which have biotransformation functions.
出处
《东南大学学报（医学版）》
CAS
2009年第5期420-423,共4页
Journal of Southeast University(Medical Science Edition)
基金
江苏省自然科学基金资助项目(BK2004012)
