差速贴壁法分选人毛囊干细胞的研究  

Enrichment of human hair follicle stem cells by adhering to typeⅣcollagen

作　　者：张群[1] 杨光辉[1] 丛笑倩[1] 崔磊[1] 张文杰[1] 刘伟[1] 曹谊林[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院整复外科,200011

出　　处：《中华实验外科杂志》2006年第6期755-757,i0002,F0003,共5页Chinese Journal of Experimental Surgery

基　　金：国家高技术研究发展计划资助项目(863计划)(2003AA205120 2003AA205122)

摘　　要：目的探讨Ⅳ型胶原差速贴壁法在分选人毛囊干细胞中的作用。方法将人毛囊ORS细胞接种在Ⅳ型胶原涂被的培养皿上进行黏附分选,实验组细胞和对照组细胞分别进行K15、β1-整合素和外皮蛋白免疫荧光和流式细胞术检查。检测细胞超微结构并测定克隆形成率。结果黏附率约为22%。实验组 K15和β1整合素的表达分别为(70．1±6．7)%和(82．4±9．3)%,对照组相应为(32．1±8．9)%和(30．3±1．3)%,外皮蛋白两组间的表达为(26．7±8．6)%和(79．2±15．3)%,两组间差异有统计学意义(P＜0．05)。电镜示实验组细胞小,核浆比例大,细胞器不成熟。实验组和对照组克隆形成率分别为(66．0±7．1)%和(30．3±2．3)%(P＜0．05)。结论利用Ⅳ型胶原差速贴壁法可初步分选、纯化人毛囊干细胞。Objective In order to enrich human hair follicle stem cells, type Ⅳ collagen coated dishes were utilized based on their different adhering potentials. Methods Human hair foUicleORS cells were cultivated on type Ⅳ collagen coated dishes. Cells were characterized by immunfluorescent stains and flow cytometry with monoclonal antibody against K15, β1-integrin and involucrin. Cell observed under electron microscopy. Colony-forming efficiency （CFE） was measured. Results About 22% adherent cells were obtained. In two groups, K15, β1 integrin and involucrin expression rate were （70.1±6.7）% and （32.1± 8.9）%, （82.4± 9.3）% and （30.3± 1.3）%, （26.7± 8.6）% and （ 79.15 ± 15.3） % respectively （ P〈0.05）. The adherent cells showed high nuclear to cytoplasmic ratio with fewer organelles. The CFE was （66.0± 7.06）% and （30.3 ± 2.3）% respectively in two groups （ P〈0.05）. Conclusion By selective adhesion to type Ⅳ collagen, human hair follicle stem cells could be enriched.

关 键 词：毛囊 干细胞 Ⅳ型胶原 黏附 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R457.1[医药卫生—基础医学]