中文摘要：

目的探讨最适合人毛囊干细胞体外培养及扩增的纤维连接蛋白（FN）铺层浓度。方法将新鲜FN稀释至所需要的4个浓度：5μg/mL、200ng/mL、10ng/mL、0ng/mL。使用酶消化法分离人毛囊细胞,将原代人毛囊干细胞在不同浓度铺层的培养皿中的细胞增值能力和克隆形成率进行统计分析,获得最适合人毛囊干细胞体外培养的铺层浓度。结果接种同一样本相同数量细胞,做克隆效率测定,在原代培养第8天做Giemsa染色,结果显示0ng/cm2铺层的培养皿中克隆团小,数量少;10ng/cm2铺层的克隆团大,数量多;200ng/cm2和5μg/cm2铺层的克隆团较小,数量不多。取10个样本,分别对4个FN铺层浓度的克隆效率进行统计分析,10ng/cm2铺层的克隆效率最高。在不同FN铺层浓度条件下,细胞扩增能力也不同,10ng/cm2铺层浓度细胞扩增能力最强,与其他3个浓度比较,差异均有统计学意义（P〈0.05）,200ng/cm2与其余3个浓度差异均无统计学意义（P〉0.05）。结论10ng/cm2的FN铺层浓度效果最佳。

英文摘要：

Objective To find the most suitable for human hair follicle stem cell adhesion of fibronectin（fibronectin,FN）concentrations.Methods The fresh fibronectin diluted to the desired concentration.The experiment is divided into the following four concentrations：5μg/mL,200ng/mL,10ng/mL,0ng/mL.Using enzyme digestion isolated from human hair follicle cells,the original generation of value-added hair follicle stem cells and cloning efficiency in a dish with different concentrations of plies,statistical analysis ply concentration most suitable for human follicle stem cells cultured in vitro.Results The same number of cells seeded in the same sample,do the cloning efficiency was measured in the first eight days of primary culture do Giemsa staining results：dish plies 0ng/cm2 of cloned groups of small,small number;10ng/cm2 Overlay dish,cloning large group and number,the smaller the dish200ng/cm2 and 5μg/cm2 plies cloned groups a small number.Take 10 samples,respectively,four FN Overlay concentration cloning efficiency statistical analysis found that,10ng/cm2 overlay genomic clone for maximum efficiency.10ng/cm2 overlay the strongest concentration of cell proliferation,and the other three groups were statistically significant,200ng/cm2 and the rest 3group of concentrations were not statistically different.Conclusion 10ng/cm2 concentration is the best FN overlay.