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目的探讨Oct4单因子转染人胎儿头皮来源的原代混杂细胞和第3代黑素细胞后,形成多能干细胞的可能性。方法培养人胎儿头皮来源的原代混杂细胞和第3代黑素细胞,应用慢病毒介导Oct4单因子诱导这些细胞。所生成的克隆采用碱性磷酸酶染色及荧光显微镜观察。结果 Oct4单因子诱导的第8和第12天,被转染的原代细胞和第3代黑素细胞出现克隆。转染后第18天,人工挑取的克隆接种于基质胶包被板,克隆很快贴壁生长,荧光显微镜下显现出绿色荧光,碱性磷酸酶染色呈紫色。结论 Oct4单因子可以诱导人胎儿头皮来源的混杂细胞及第3代黑素细胞产生克隆,但产生效率很低。
Objective To investigate the possibility of pluripotent stem cells after Oct4 single factor transfection of human primary scalp derived primary mixed cells and 3rd generation melanocytes. Methods Primary mixed scFv cells and third generation melanocytes were cultured and induced by lentivirus. The generated clones were stained with alkaline phosphatase and observed under a fluorescence microscope. Results On the 8th and 12th days induced by Oct4 single factor, the primary cells transfected and the 3rd generation melanocytes appeared clones. On the 18th day after transfection, the clones picked by hand were inoculated on the matrigel coated plate, the clones grew rapidly adherently, the green fluorescence appeared under the fluorescence microscope, and the alkaline phosphatase staining was purple. Conclusion Oct4 single factor can induce clonal hybridization of human fetal scalp and third generation melanocytes, but the production efficiency is very low.