表皮干细胞的分选鉴定及培养

Screening,evaluation and in vitro cultivation of epidermal stem cells

目的探讨并评价Ⅳ型胶原黏附法分离表皮干细胞的可行性。方法以不同浓度Ⅳ型胶原及不同黏附时间对表皮细胞进行分选培养。选择0.2mg/ml、15min组细胞,进行免疫细胞化学染色鉴定。结果黏附浓度对贴壁率影响不明显,而黏附时间的影响显著;0.2mg/mlⅣ型胶原黏附15min的细胞,β1整合素、K19免疫荧光细胞化学双重标记阳性细胞的比例约为28.50%。结论Ⅳ型胶原黏附法分选表皮干细胞仅仅是一种初步的筛选,是目前组织工程产业化种子细胞的分选最可行的方法,但要获取高纯度的ESC需要进一步的深入研究。

Objective To discuss the feasibility of epidermal stem cells（ESC） dissociation by means of type Ⅳ collagen adhesiveness. Methods ESC was screened and cultured in type Ⅳ collagens with different concentrations at different adhesiveness time points. ESC（0.2 mg/ml） at 15 minutes was selected for immunocytochemical stain evaluation. Results The adhesiveness concentration exerted insignificant effect on anchoring rate but adhesiveness time significantly affected anchoring rate. In ESC adhered with type Ⅳ collagen at 0.2 mg/ml for 15 minutes, percentage of double immunofluorescence cytochemical stained positive cells of β1 integrin to K19 was 28.50% or so. Conclusion Type Ⅳ collagen adhesiveness screening ESC is only a primary step but the most feasible way for screening of tissue-engineered industrial seed cells. However,a further study is needed to obtain high purity ESC.