中文摘要：

目的 观察转化生长因子-β1（TGF-β1）对体外培养的毛囊干细胞（HFSCs）生物学特性的影响.方法 体外分离培养人HFSCs,观察其在10-μg/L TGF-β1条件培养基下的细胞形态变化,噻唑蓝（MTT）比色法于24、48、72 h分别检测加入条件培养基前后细胞增殖能力,细胞接近铺满时划痕,并于0、24、48、72 h后用目镜测微尺测量划痕宽度检测迁移能力,将细胞接种于Transwell小室上层,下层加入含10 μg/L TGF-β1条件培养基,48 h后计数穿透细胞数,免疫荧光化学方法检测加入条件培养基前后角蛋白19、β-整合素、E钙黏蛋白、N钙黏蛋白、波形蛋白的表达.结果在TGF-β1作用下,HFSCs之间的紧密连接消失,细胞由圆形或多角形向梭形细胞转变,MTT结果显示,在TGF-β1作用下HFSCs各时间点A值与对照组比较,差异无统计学意义（P〉0.05）,细胞划痕实验结果显示,TGF-β1能够促进细胞迁移,对照组差异有统计学意义（P〈0.01）.Transwell穿透实验结果表明,加入TGF-β1后,HFSCs细胞定向迁移穿透基底膜数量与对照组比较显著增多（47.9±8.8比35.8±6.7,P〈0.01）,免疫荧光结果显示TGF-β1能够降低角蛋白19、β-整合素和E钙黏蛋白表达,而N钙黏蛋白、波形蛋白的表达增加.结论 TGF-β1通过诱导体外培养的HFSCs上皮间质转化而促进其迁移.

英文摘要：

Objective To investigate the effects of transforming growth factor （TGF）-β1 on biological activities of hair follicle stem cells （HFSCs） in vitro.Methods The morphological and biological characters of HFSCs were observed when cultured with keratinocyte-serum free medium （K-SFM） containing 10μg/L TGF-β1 and K-SFM in vitro.The proliferative abilities were examined by MTT assay at the time points of 24,48,96 h after the HFSCs were cultured with both medium.The migration ability was measured by wound healing assay at the time points of 0,24,48,72 h.The penetrating cells were calculated at the 48th h after cultured in Transwell with both medium,and the expression of K19,β-intergrin,E-Cadhenin,N-Cadhenin and Vimentin were also detected by immunofluorescence.Results The HFSCs cultured with TGF-β1 lost their typical cobblestone patterns of epithelial cells,displaying spindle-shape,fibroblast-like morphology.The absorbance （A） of HFSCs in each time point had no significant difference between two groups （P〉0.05）.The wound healing assay showed that the migration ratio of HFSCs induced by TGF-β1 was higher than in control group （P 〈0.01 ）,and the counts of HFSCs penetrating through membrane of Transwell in TGF-β1 group were 47.9 ± 8.8,significantly more than those in the control group [（ 35.8 ± 6.7 ）,P〈0.01].Immunofluoresence revealed that the expression of K19,β-intergrin and E-Cadhenin was decreased and that of N-Cadhenin and Vimentin was increased in cells stimulated by TGF-β1.Conclusion TGF-β1 can prompt HFSCs migration activity by driving their epithelial-mesenchymal-transition in vitro.