大鼠表皮干细胞的分布与体外分离培养  

Distribution, isolation and culture of epidermal stem cells of rats in vitro

作　　者：陈伟[1] 杨恬[1] 连小华[1] 杨珂[1] 黄恩毅[2] 

机构地区：[1]第三军医大学基础医学部细胞生物学教研室,重庆400038 [2]重庆大学生物工程学院,重庆400044

出　　处：《第三军医大学学报》2007年第5期376-379,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30371383)~~

摘　　要：目的研究大鼠表皮干细胞的分布与体外分离培养。方法用免疫组织化学技术研究大鼠表皮干细胞的分布;用DispaseⅡ、胰酶二步法消化新生大鼠皮肤,获得单细胞悬液,Ⅳ型胶原筛选,体外培养大鼠表皮干细胞,进行免疫组化鉴定,荧光标记流式细胞仪分析。结果在表皮基底层、毛囊外根鞘区α6-integrin、K15等表皮干细胞标记物染色阳性,而分化指标CD71染色阴性;CD34在表皮基底层阴性表达,而在毛囊隆突区阳性表达;体外分离的基底层表皮干细胞生长良好,具有较高的克隆形成率;干细胞标记物α6-integrin、K15呈阳性表达,流式细胞仪检测α6-integrin达到84%,说明较为成功地体外分离培养出大鼠表皮基底层干细胞。结论在表皮基底层、毛囊外根鞘区存在表皮干细胞。Objective To investigate the distribution, isolation and culture of the epidermal stem cells from rats. Methods Immnohistochemical methods were used to confirm the location of the epidermal stem cells. The skin of neonatal rats were dissociated into single cells by dispase Ⅱ and trypsin solution, the rapidly adherent cells to collgenⅣ were cultured with KSFM ,and those no rapidly adherent cells were regarded as control. Immunohistology and flow cytometry were conducted to identify the epidermal stem cells. Results α 6integrin and K15 were expressed in the basal layer cells and hair follicle bulge cells, while the CD71 was negative negatively expressed. CD34 were expressed in the hair follicle bulge cells while not in the basal layer cells. The epidermal stem cells isolated by collgenⅣ had higher colony forming efficiency. Immunocytochemical staining showed that α 6-integrin and K15 were strongly expressed in the cultured epidermal stem cells. Flow cytometry indicated that 84% cultured epidermal stem cells were expressed α 6-integrin. Conclusion The epidermal stem cells of rats are located at the basal layer of epidermis and the hair follicle bulge.

关 键 词：表皮干细胞 细胞培养 体外 

分 类 号：R322.991[医药卫生—人体解剖和组织胚胎学] R329.2[医药卫生—基础医学]