Isolation, culture and identification of hmnan epithelial stem cells

LI Dan; et al

doi:10.13406/j.cnki.cyxb.2007.12.002

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Isolation, culture and identification of hmnan epithelial stem cells

更新时间：2024-01-17

- Isolation, culture and identification of hmnan epithelial stem cells
- Journal of Chongqing Medical University Issue 12, Pages: 1272-1275(2007)
- 作者机构：
  
  1. 第三军医大学西南医院整形外科
  2. 第三军医大学西南医院整形外科,重庆,400038
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2007.12.002
  CLC： R329
- Published：2007，
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[1]李丹,李世荣,曹川.人表皮干细胞的体外分离培养和鉴定[J].重庆医科大学学报,2007(12):1272-1275.

LI Dan, et al. Isolation, culture and identification of hmnan epithelial stem cells. [J]. Journal of Chongqing Medical University (12):1272-1275(2007)
[1]李丹,李世荣,曹川.人表皮干细胞的体外分离培养和鉴定[J].重庆医科大学学报,2007(12):1272-1275. DOI： 10.13406/j.cnki.cyxb.2007.12.002.

LI Dan, et al. Isolation, culture and identification of hmnan epithelial stem cells. [J]. Journal of Chongqing Medical University (12):1272-1275(2007) DOI： 10.13406/j.cnki.cyxb.2007.12.002.

摘要

目的:探讨人表皮干细胞的体外快速分离培养及鉴定方法。方法:中性蛋白酶和胰蛋白酶两步法从手术切除的人包皮组织中分离表皮层和真皮层

并获得表皮单细胞悬液

采用Ⅳ型胶原铺板选择性黏附、分离和角质细胞无血清培养基（K-SFM）培养表皮干细胞。倒置显微镜下观察培养细胞的生长状况

检测细胞克隆形成率

免疫组化染色观察表皮干细胞标志物β1整合素和角蛋白19（K19）的表达;以角质形成细胞作为对照。结果:组织学观察显示

培养24h后细胞呈克隆状生长;所分离、培养细胞的克隆形成率高于对照角质形成细胞;免疫组化染包显示

培养细胞β1整合素及K19均呈阳性表达。结论:运用Ⅳ型胶原黏附结合K-SFM培养可以实现人表皮干细胞的体外快速分离和培养。

Abstract

Objective:To explore a method for isolation and culture of human epidermal stem cells. Methods:The Epidermal stem cell were isolated by adhering to collagen typeⅣafter obtained by digesting human foreskin with DispaseⅡand Tryp and culture in vitro in K-SFM. The expressions ofβ1-integrin and keratin 19 (K19) in epidermal stem cell were detected with immunocytochemical methods

and the colony forming efficiency was also studied. Keratinocytes were served also detected. Results:It was revealed by histologieal observation that colonieswere formed 24 hours after inoculation. The isolated and cultured cell cloning efficiency was higher than that of the control group. Positive expression ofβ1-integrin and K19 of cultured cells was detected by immunocytochemistry. Conclusion:Adult epidermal stem cells could be successfully isolated and cultured by adhension with typeⅣcollagen and culture with K-SFM

关键词

表皮干细胞细胞培养细胞分离鉴定

Keywords

Epidermal stem cellscell culturecell isolationidentification

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