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目的:探讨并建立表皮干细胞Oct4基因的非转基因诱导方法。方法:分离培养人表皮干细胞,在培养至3~5d出现克隆性生长时加入工作浓度为10μmol/L的5-AzaC,24h后再诱导1次。采用RT-PCR方法,检测经5-AzaC刺激48h后的表皮干细胞Oct4基因转录水平。结果:PCR产物经1%琼脂糖凝胶电泳后,出现247bp(Oct4)和219bp(β-actin)目的片段明亮的特异性条带。结论:5-AzaC能有效激发出表皮干细胞Oct4基因的转录。
Objective: To explore and establish a non-transgenic induction method of Oct4 gene of epidermal stem cells. Methods: Human epidermal stem cells were isolated and cultured. 5-AzaC at the concentration of 10μmol / L was added to the clonogenic cultures after 3-5 days of culture, and then induced once more after 24 hours. The transcription level of Oct4 gene in epidermal stem cells after 5-AzaC stimulation was detected by RT-PCR. Results: The specific bands of 247bp (Oct4) and 219bp (β-actin) fragments were detected by 1% agarose gel electrophoresis. Conclusion: 5-AzaC can effectively stimulate the transcription of Oct4 gene in epidermal stem cells.