体外定向诱导人骨髓基质干细胞内皮化  

Inducement of Endothelial Differentiation in Human Bone Marrow Stromal Cells In Vitro

作　　者：胡昕婴[1] 史剑慧[1] 牛玉宏[1] 葛均波[1] 

机构地区：[1]复旦大学附属中山医院心内科上海市心血管病研究所,上海200032

出　　处：《复旦学报（医学版）》2004年第4期336-339,F002,共5页Fudan University Journal of Medical Sciences

基　　金：上海市医学发展基金-重点研究课题(2 0 0 0I-2D0 0 2 )资助项目

摘　　要：目的　探索体外定向诱导人骨髓基质干细胞 (humanbonemarrowstromalcell,hMSC)内皮分化的潜能与条件。方法　采用含多种生长因子的内皮细胞支持液EGM2 MV(Clonetics)作为体外内皮化诱导剂 ,观察形态学和细胞表面免疫标志的变化 ,研究hMSC体外内皮化的条件和诱导效果。结果　经 2 5 %EGM 2 MV诱导后 ,细胞生长良好 ,在多种生长因子的刺激下 ,原先长梭形的细胞缩短 ,出现鹅卵石样形态。诱导培养 10d后 ,经流式细胞学检测 ,内皮干细胞标志CD133阳性率明显升高 ,基质干细胞标志CD10 6阳性率下降。结论　hM SC具有向血管内皮细胞分化的潜能。利用含有多种生长因子的内皮细胞支持液EGM2 MV ,可在体外诱导hMSC向内皮细胞分化。本研究为利用自体MSC移植重建心脏血运提供实验依据。Purpose: The inducing capability of EGM2-MV(Clonetics), an endothelial sustaining medium containing human epidermal growth factor(hEGF), vascular endothelial growth factor(VEGF), R3 insulin-like growth factor-1(R3-IGF-1, an analogue of IGF-1), human fibroblast growth factor-B(hFGF-B), hydrocortisone and ascorbic acid, on hMSC endothelial development in vitro was investigated in this study. Under the stimulation of these growth factors, hMSC was triggered to vascular endothelial differentiation. Methods: To optimize the inducing condition, hMSC was cultured in basal medium with different concentration of EGM2-MV(25%, 50%, 100%). We investigated the morphological change after the inducement. Cell phenotype characteristics were analyzed through FACS before and after the treatment of EGM2-MV to confirm the differentiation of endothelium. Results: The cells cultured in 100% and 50% of EGM2-MV were found to be suspending in the medium 3 days after culturing. Observed through an inverted-type phase-contrast microscope, the cells were swelling and splitting. While hMSC cultured in 25% of EGM2-MV was growing sound. The former long fusiform cell decreased in length and displayed a single-layered cobble-like morphology. Through FACS analysis we found that hMSC could be induced to increase the expression of endothelial phenotype characteristic CD133 under the stimulation of the growth factors. Meanwhile the expression of CD106, one of mesenchymal cell markers, was decreased significantly. Conclusions: The results showed that hMSC could be triggered to endothelial differentiation by being cultured in adequate concentration of endothelial sustaining medium EGM2-MV in vitro. Under the stimulation of the growth factors, hMSC could be induced to display a cobble-like appearance and express endothelial phenotype characteristics, indicating that hMSC represents a promising cell source for cardiac revascularization.

关 键 词：体外定向诱导 人骨髓基质干细胞 内皮化 内皮细胞 表面标志 

分 类 号：R392.1[医药卫生—免疫学]