中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (10): 1813-1816.doi: 10.3969/j.issn.1673-8225.2012.10.023

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

乙醛脱氢酶活性检测子宫内膜干细胞★

张晓露，何援利，马  颖   

  1. 南方医科大学珠江医院妇产科，广东省广州市 510282
  • 收稿日期:2011-10-08 修回日期:2011-12-28 出版日期:2012-03-04 发布日期:2012-03-04
  • 通讯作者: 何援利，硕士，教授，博士生导师，南方医科大学珠江医院妇产科，广东省广州市 510282 heyuanli310@yahoo.com
  • 作者简介:张晓露★，女，1987年生，汉族，南方医科大学在读硕士，主要从事妇科的研究。zhangxiaolu1333@sina.com

Application of acetaldehyde dehydrogenase in detection of endometrial stem cells

Zhang Xiao-lu, He Yuan-li, Ma Ying   

  1. Department of Obstetrics and Gynecology, Zhujiang Hospital of Southern Medical University, Guangzhou  510282, Guangdong Province, China
  • Received:2011-10-08 Revised:2011-12-28 Online:2012-03-04 Published:2012-03-04
  • Contact: author: He Yuan-li, Master, Professor, Doctoral supervisor, Department of Obstetrics and Gynecology, Zhujiang Hospital of Southern Medical University, Guangzhou 510282, Guangdong Province, China heyuanli310@yahoo.com
  • About author:Zhang Xiao-lu★, Studying for master’s degree, Department of Obstetrics and Gynecology, Zhujiang Hospital of Southern Medical University, Guangzhou 510282, Guangdong Province, China zhangxiaolu1333@sina.com

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摘要：

背景：人子宫内膜中存在干细胞特性的细胞，但特异性标志物尚未找到，导致此类细胞很难被分离。
目的：分析乙醛脱氢酶(aldehyde dehydrogenase，ALDH)活性检测能否分选出子宫内膜干细胞。
方法：采用机械和酶消化相结合方法分离人子宫内膜细胞，用两次滤网方法分离出基质细胞和上皮细胞，根据乙醛脱氢酶活性采用流式细胞仪分选出ALDHhigh细胞和ALDHlow细胞，用有限稀释法培养细胞，观察细胞形态及生长特性；采用流式细胞仪鉴定细胞c-kit，CD90，CD73及CD29的表达。
结果与结论：基质细胞中ALDHhigh细胞占(1.88±0.17)%，经过15 d的原代培养后，ALDHlow细胞克隆形成率达(1.06±0.34)%，ALDHhigh细胞克隆形成率达(4.50±0.82)%，ALDHhigh细胞集落形成率较ALDHlow细胞高(P < 0.05)，同时ALDHhigh细胞大克隆数较ALDHlow高(P < 0.05)；上皮细胞中未发现ALDHhigh细胞及克隆形成。子宫内膜集落形成细胞表现出c-kit、CD29、CD90、CD73阳性。说明ALDH活性检测对子宫内膜干细胞有一定的分选作用。

关键词: 人子宫内膜干细胞, 成体干细胞, 乙醛脱氢酶, ALDHhigh细胞, 有限稀释法, 表型鉴定 

Abstract:

BACKGROUND: Stem-like cells have been found in human endometrium but the specificity marker has not been found. This kind of cells is difficult to be separated.
OBJECTIVE: To investigate the usefulness of aldehyde dehydrogenase (ALDH) activity as a marker for endometrial stem cells.
METHODS: Human endometrial cells were isolated and dissociated mechanically and enzymatically from human endometrium. Stromal and epithelial cells were separated by two series of filters. The ALDHhigh cells and ALDHlow cells were selected by flow cytometry based on ALDH activity. Limiting dilution culture was performed to culture these cells. Cell morphology and growth characteristics were observed. Flow cytometry analysis was used to identify the expression of c-kit, CD90, CD73 and CD29 markers in cultured cells.
RESULTS AND CONCLUSION: The proportion of ALDHhigh cells was (1.88±0.17)% in stromal cells. After primary cultured for 15 days, the colony forming rate of the ALDHhigh cells reached 4.50%±0.82% which were significantly higher than ALDHlow cells (1.06±0.34)% (P < 0.05). Meanwhile, the large colony forming number of ALDHhigh cells was higher than that of ALDHlow cells (P < 0.05). The ALDHhigh cells and colony forming were not detected in epithelial cells. The expressions of c-kit, CD29, CD90 and CD73 cells were positive in endometrium colony forming cells. Results have proved that ALDH activity detection can select the endometrial stem cells.

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