第三代端粒酶缺失小鼠皮肤表皮干细胞分离培养特性观察

【摘要】目的 观察第三代端粒酶缺失小鼠与野生型小鼠皮肤表皮干细胞数目及功能差异。方法 利用流式细胞术对皮肤表皮干细胞进行数目分析及分选；实时定量PCR方法检测p21基因表达；表皮干细胞克隆形成数目检测细胞自我更新能力。结果 野生型小鼠基底部表皮干细胞与基底部上层表皮干细胞比例分别为9.56% 和1.22%，在第三代端粒酶缺失小鼠中这两部分细胞比例分别为17.36% 和 2.92%；第三代端粒酶缺失小鼠p21相对表达水平为野生型小鼠的6.4倍；野生型小鼠每104 个表皮干细胞能够形成280.2个克隆，第三代端粒酶缺失小鼠每104 表皮干细胞仅能够形成29.28个克隆,以上结果均具有统计学差异（p<0.05）。结论 与野生型小鼠比较，第三代端粒酶缺失小鼠皮肤表皮干细胞出现衰老性的损伤。

Isolation and Culture of Epidermal Stem Cells from the Third Generation of Mice That Were Knocked-out ofTerc

Abstract] Objective To observe the difference in epidermal stem cell number and function between wild-type (WT) mice and the third generation of Terc knockout (G3Terc-/-) mice. Methods Flowcytometry was used to analyse and sort epidermal stem cells;Quantitative real-time PCR to analyse the relative expression level of p21 in epidermal stem cells; The ability of self-renewal was reflected by the number of colonies formed by epidermal stem cells. Results Ratio of basal and suprabasal epidermal stem cells in WT mice were 9.56% and 1.22% respectively, ratio of basal and suprabasal epidermal stem cell in G3Terc-/- mice were 17.36% and 2.92%. Relative p21 expression level in G3Terc-/- mice was 6.40 fold to WT mice; Number of colonies formed by WT mice epidermal stem cells were 280.2 per 104cell,number of colonies formed by G3Terc-/- mice epidermal stem cells were 29.28 per 104 cell, it has significant difference in the above results（p<0.05）.Conclusion Compared to WT mice, epidermal stem cells in G3Terc-/- mice were aging.