目的探讨体外诱导人毛囊干细胞成血管平滑肌细胞的可行性。方法采用中性蛋白酶(Dispase)分离人毛囊干细胞,用含10 ng/mL PDGF-BB、10%血清的低糖DMEM诱导液对其进行诱导,无PDGF-BB的培养液为对照组,观察每代细胞形态,诱导4代后检测α-平滑肌肌动蛋白(α-SM actin)与肌钙结合蛋白(Calponin)的表达。结果在诱导液的作用下,细胞形态逐步向平滑肌样转变,对照组细胞形态改变不显著。细胞免疫荧光检测显示,至第4代,实验组已明显表达α-SM actin与Calponin,对照组表达不明显;流式细胞仪检测显示,实验组α-SM actin与Calponin阳性表达率近50%,对照组则低于8%;RT-PCR显示,实验组表达Calponin,而对照组未见明显表达。结论使用含10 ng/mLPDGF-BB的低糖DMEM培养液可体外诱导人毛囊干细胞成血管平滑肌细胞。

Objective To investigate the feasibility of inducing human hair FSCs(follicle stem cells) into VSMCs(vascular smooth muscle cells) in vitro.Methods The human hair FSCs were isolated by dispase and cultured in DMEM containing 10 ng/mL PDGF-BB as experimental group,and the human hair FSCs cultured in DMEM without PDGF-BB were set up as control group.The morphological change of each passage was observed,the expression of α-Smooth Muscle actin(α-SM actin) and calponin in each group were tested after inducing FSCs at passage 4.Results The VSMC-like change was obviously observed in the induced FSCs,while the same phenomenon was not found in control group.α-SM actin and calponin were expressed by immunocytochemical staining in the induced FSCs of fourth passage,while little expression was observed in the control group.FCM showed nearly 50% of expression rate of α-SM actin and calponin in the experimental group,much higher than 8% in control group.RT-PCR further confirmed the expression of calponin in experimental group,and no obvious expression of calponin was tested in control group.Conclusion FSCs can be induced into VSMCs in vitro by DMEM containing 10 ng/mL PDGF-BB.