人表皮干细胞体外诱导分化为汗腺样上皮细胞的研究

发布时间：2018-04-22 14:49

本文选题：表皮干细胞 + 汗腺样细胞　；参考：《重庆医学》2017年10期

【摘要】：目的探讨体外人表皮干细胞向汗腺样上皮分化的影响条件,并对诱导成的管腔样结构进行鉴定。方法将人表皮干细胞接种到复方壳多糖真皮基质上和胶原凝胶中,加入不同浓度的表皮生长因子(EGF),体外垂直震荡培养,进行三维培养和定向诱导分化,采用HE染色、免疫荧光等手段观察表皮干细胞定向分化为汗腺样上皮的条件及形态、表型改变。结果 15~20ng/mL的EGF可诱导组织工程真皮上的表皮干细胞向真皮内生长并可出现腺样结构。HE染色该结构,显示为单层细胞连接为环状,中间见明显的腔隙,细胞质嗜酸性。利用CK19荧光染色,在激光共聚焦显微镜下观察到细胞团中央有管腔结构,同时此结构表达CK18、癌胚抗原(CEA)。结论体外培养的人表皮干细胞接种在组织工程真皮上,在一定浓度的EGF诱导条件下,能形成管腔样结构,该管腔样结构在形态学、组织学上与在体汗腺分泌部细胞相似。
[Abstract]:Objective to investigate the effect of human epidermal stem cells on the differentiation of human epidermal stem cells into sweat gland epithelium in vitro and to identify the induced luminal structure. Methods Human epidermal stem cells were inoculated into compound chitosan dermal matrix and collagen gel. Different concentrations of epidermal growth factor (EGFN) were added to culture in vitro vertical shock culture, three-dimensional culture and directional differentiation, HE staining was used. The morphological and phenotypic changes of epidermal stem cells differentiated into sweat gland-like epithelium were observed by immunofluorescence. Results 15~20ng/mL EGF could induce the epidermal stem cells to grow into the dermis, and the glandular structure. He staining showed that the monolayer cells were connected to each other in annular shape, with obvious lacunae and eosinophilic cytoplasm. By using CK19 fluorescence staining, the lumen structure in the center of the cell was observed under laser confocal microscope, and the structure expressed CK18, carcinoembryonic antigen (CEA). Conclusion cultured human epidermal stem cells inoculated on tissue engineered dermis can form luminal structure under certain concentration of EGF induction. The morphology and histology of human epidermal stem cells are similar to those of sweat gland secretory cells in vivo.
【作者单位】：第三军医大学大坪医院野战外科研究所整形美容科;第三军医大学大坪医院野战外科研究所皮肤科;
【基金】：国家自然科学基金资助项目(81201244)
【分类号】：R622

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