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  

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

作　　者：张俊伶[1] 黄颂[1] 路璐[1] 李德冠[1] 吴红英[1] 孟爱民[1] 

机构地区：[1]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,300192

出　　处：《天津医药》2014年第8期762-764,共3页Tianjin Medical Journal

基　　金：国家973重大专项资助项目(2011CB964800-G);国家自然科学基金海外合作基金(81129020);国家自然基金面上项目(81072237;81372928);天津市重点基金资助项目(11JCZDJC19100);教育部高等学校博士学科点专项基金(20111106110038)

摘　　要：目的观察第三代端粒酶缺失(G3Terc-/-)小鼠与野生型(WT)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 WT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±1.06)%和(1.22±0.08)%,在G3Terc-/-小鼠中这两部分细胞比例分别为(17.36±3.56)%和(2.92±0.72)%;G3Terc-/-小鼠p21相对表达水平为WT小鼠的6.4倍;WT小鼠每104个表皮干细胞能够形成(280.20±29.81)个克隆,G3Terc-/-小鼠每104表皮干细胞仅能够形成(29.28±5.24)个克隆,以上结果差异均有统计学意义(均P<0.05)。结论与WT小鼠比较,G3Terc-/-小鼠皮肤表皮干细胞出现衰老性的损伤。Objective To observe the difference in number of epidermal stem cell and its 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 is used to analyse the relative expression level of p 21 in epider-mal stem cells;Self-renewal ability was reflected by the number of colonies formed by epidermal stem cells. Results Basal and suprabasal ratios in epidermal stem cells in WT mice were （9.56 ± 1.06）% and （1.22 ± 0.08）% respectively; basal and suprabasal ratios in epidermal stem cell in G3Terc-/-mice were （17.36±3.56）%and （2.92±0.72）%respectively. 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 （280.20±29.81） per 104 cell, number of colonies formed by G3Terc-/-mice epidermal stem cells were （29.28±5.24） per 104 cell, which present significant difference to each other（P〈0.05）. Conclusion Compared to WT mice, epidermal stem cells in G3Terc-/-mice were aging.

关 键 词：小鼠 基因敲除 皮肤衰老 干细胞 端粒 末端转移酶 

分 类 号：Q255[生物学—细胞生物学]