摘要
目的:研究表皮干细胞在成人体皮中的分布及其分离和培养。方法:用免疫组织化学方法检测成人体皮中β1整合素及角蛋白19的表达,对成人体皮中的表皮干细胞进行定位和定量;用Ⅳ型胶原分离培养成人体皮表皮干细胞,通过检测β1整合素及角蛋白19的表达水平及克隆形成率(以角质细胞为对照)对其进行鉴定。结果:β1整合素及角蛋白19的免疫组化阳性信号主要位于表皮的基底部及毛囊周围,阳性细胞在表皮中所占的数量较少,且在真皮中也有少量表达。体外培养,可见细胞呈克隆样生长、β1整合素及角蛋白19免疫组织化学染色呈阳性,且其克隆形成率为17.17％,明显高于对照组的6.83％(P<0.05)。结论:应用Ⅳ型胶原快速贴附法及成人成纤维细胞条件培养液,对成人体皮中表皮干细胞成功进行了分离培养,为表皮干细胞体外的大量扩增奠定了基础。
Objective:To study the localization and in -vitro culture of epidermal stem cells in normal adult skin. Meth ods:The expressions of β1 integrin and keratin 19(K19) in normal adult skin were detected with immunocytochemical methods. The epidermal stem cells were isolated by adhering to collagen type IV and cultured in vitro in conditional medium prepared according to that for human fibroblasts. Then, the cultured cells were identified by immunohistochemistry staining of (3, integrin and K19, and the colony forming efficiency was also studied. Gliocytes were taken as control. Results: The cells expressing integrin β1 and K19 in epidermal layer were less. The epidermal stem cells selected by rapid adherence to type Ⅳ collagen formed large colonies after 10 d. The clone forming rate of epidermal stem cells was 17. 17% , which was much higher than that in control group (6. 83% ,P<0. 05). Conclusion: The adult epidermal stem cells can be isolated in vitro by means of rapid adherence to type Ⅳ collagen, and can be cultured with conditional medium.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2004年第8期815-817,共3页
Academic Journal of Second Military Medical University
基金
上海市科委重大研究计划(03DJ140203).
关键词
表皮干细胞
细胞培养
细胞分离
epidermal stem cells
cell culture
cell separation
