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目的 探讨胚胎干细胞 (ES)源表皮样干细胞的分化潜能 ,为研究其分化的调控及寻找新的皮肤组织工程种子细胞奠定基础。 方法 将Hoechst33342标记或未标记的小鼠ES细胞 ,与人羊膜共培养 4d ,诱导其定向分化为表皮样干细胞克隆 ,胰酶消化后移植于裸鼠皮下 ,10、2 0、30和 4 5d ,对移植后细胞的分化进行形态学和免疫组织化学观察分析。 结果 10～ 2 0d后 ,标记的细胞可分化为由单层或复层上皮样细胞构成的管状或泡状结构 ,它们分别呈 β1 整合素、CK19、CK15、PK、套膜蛋白 (involucrin)和CEA阳性 ;移植 4 5d后 ,可见角化复层扁平上皮、皮脂腺样、汗腺样和毛囊样等组织结构。 结论 小鼠ES源表皮样干细胞具有分化为角化复层扁平上皮和毛囊样、汗腺样和皮脂腺样等结构的潜能。
Objective To investigate the differentiation potential of embryonic stem cell (ES) -derived epidermal-like stem cells and lay the foundation for the study of differentiation regulation and search for new skin tissue engineering seed cells. Methods Hoechst33342 labeled or unlabeled mouse ES cells were co-cultured with human amniotic membrane for 4 days to induce their differentiation into epidermal-like stem cell clones. After being digested with trypsin, they were subcutaneously transplanted in nude mice at 10, 20, 30 and 45 days, The morphological and immunohistochemical analysis of the differentiated cells after transplantation was performed. Results After 10 ~ 20d, the labeled cells could differentiate into tubular or vesicular structures composed of monolayer or stratified epithelial-like cells. They were β1 integrin, CK19, CK15, PK, involucrin and CEA positive; 4d after transplantation, we can see keratinized stratified squamous epithelium, sebaceous gland, sweat gland-like and hair follicles-like tissue structure. Conclusion Mouse ES-derived epidermal-like stem cells have the potential to differentiate into keratinized flattened epithelium and hair follicle-like, sweat-like and sebaceous-like structures.