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.docVIP

  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# 张俊伶，路璐，李德冠，吴红英，孟爱民** 5 10 15 20 25 30 35 40 （中国医学科学院放射医学研究所，天津市放射医学与分子核医学重点实验室） 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 10000 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（p0.05）。 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。 关键词：皮肤生理学 野生型小鼠 第三代端粒酶缺失小鼠 皮肤表皮干细胞 衰老 中图分类号：Q419 Observation of isolation and culture in the third generation of Terc knockout mice epidermal stem cell ZHANG junling, LUlu, LI deguan, WU hongying, MENG aimin (Tianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine, Institute of Radiation Medicine,Chinese Academy of Medical Sciences) 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. Method 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. Result 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（p0.05）.Conclusion Compared to WT mice, epidermal stem cells in G3Terc-/- mice were aging. Key words: Dermatophysiology ;Wild type mice;The third generation of Terc knockout (G3Terc-/-) mice; Epidemal stem cell;Aging 0 引言