毛囊干细胞体外诱导分化为血管内皮细胞的实验研究

目的探讨体外诱导人毛囊干细胞成血管内皮细胞的可行性。方法采用中性蛋白酶(Dispase)分离人毛囊干细胞,用含10 ng/mL VEGF、2 ng/mL bFGF及10%血清的EGM-2诱导液对其诱导,以无诱导因子的基础培养液为对照组,对每代细胞形态进行观察。诱导4代后,检测vWF(von Willebrand factor)与CD31的表达。结果在诱导液的作用下,细胞形态逐步向内皮细胞的铺路石样形态转变,对照组细胞形态改变不明显。至第4代,实验组已明显表达vWF与CD31,对照组表达不明显;流式细胞仪检测显示,实验组阳性表达率近80%,对照组则低于5%;RT-PCR显示,实验组表达vWF,对照组未见明显表达。结论使用含10 ng/mL VEGF、2 ng/mL bFGF及10%血清的EGM-2诱导液,可在体外诱导人毛囊干细胞分化成血管内皮细胞。

Preliminary Study on Inducing Hair Follicle Stem Cells into Vascular Endothelial Cells in Vitro

Objective To investigate the feasibility of inducing human hair FSCs （follicle stem cells） into VECs （vascular endothelial cells） in vitro. Methods The dispase was used to isolate the stem cells and stem cells were cultured in EGM-2 plus 10 ng/mL VEGF and 2 ng/mL bFGF as long as 4^th passage as a experimental group, the control group was cultured in EGM-2 without growth factor. The morphological change of each passage was observed and vWF （von Willebrand factor） and CD31 were tested 4 passages later. Results The morphological change to VECs was obviously found in the increasing passage of the induced FSCs, but the phenomenon was not distinct in the control group. Induced FSCs of 4^th passage expressed vWF and CD31 in immunocytochemical staining, while little expression was observed in the control groups. FCM showed nearly 80% of induced FSCs expressed vWF and CD31, much higher than the control ones which were lower 5%. RT-PCR further confirmed the experimental group expressed vWF, but not obvious expression in the control group. Conclusion FSCs can be induced to VECs in vitro by EGM-2 plus 10 ng/mL VEGF and 2 ng/mL bFGF.