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异位子宫内膜干细胞的检测及其标志物研究

王宇全1,尹利荣1,郭蕊萌2   

  1. 1. 天津医科大学第二医院妇科
    2. 天津医科大学第二医院
  • 收稿日期:2012-06-26 修回日期:2012-12-15 出版日期:2013-05-15 发布日期:2013-05-15
  • 通讯作者: 王宇全

The Study of Identification of Ectopic Endometriail Stem Cells and Exploxion of Its Markers in Mouse Model of Endometriosis.

WANG Yuquan ,YIN Li rong,GUO Ruimeng   

  1. Department of Gynecology, The Second Hospital of Tianjin Medical University
  • Received:2012-06-26 Revised:2012-12-15 Published:2013-05-15 Online:2013-05-15
  • Contact: WANG Yuquan

摘要:

【摘要】 目的  了解异位子宫内膜干细胞在病灶中的数量、分布及定位情况,并筛选其可能的标志物表达。方法 在子宫内膜异位症裸鼠模型建造前24h开始进行5-溴脱氧尿苷(BrdU)脉冲标记,共5d。在标记结束后4h、1周、3周、5周、6周、7周、8周及9周,通过免疫组化检测异位病灶中BrdU的表达情况,确定标记滞留细胞(The label-retaining cells,LRCs)。通过免疫组化双染检测LRCs表面CD34和CD146的表达情况。结果  腺上皮和间质的BrdU初始最大标记率分别为(69.89±0.78)%和(73.27±1.06)%,于第6周和8周时分别有约(2.07±0.19)%和(4.63±0.72)%的腺上皮和间质细胞仍保留标记,即为上皮和间质标记滞留细胞,多位于异位内膜与所种植的裸鼠组织的交界处。LRCs并不表达CD34,少数间质LRCs表达CD146。结论 异位子宫内膜中有上皮和间质两种干细胞,主要定位于异位内膜与所种植的组织的交界处,CD34不是异位内膜干细胞的标志物,CD146表达于异位内膜干细胞,但并非其特异性标志物。 

关键词: 子宫内膜异位症, 干细胞, 染色与标记, 溴脱氧尿苷, 免疫组织化学, 抗原, CD34, 抗原

Abstract: Abstract   Objective  To identify and locate adult stem cells in ectopic endometrium and to find its markers using the 5-bromo-2-deoxyuridine (BrdU) labeling method in mouse model of endometriosis. Method  The female nude mice were pulse labeled by BrdU for 5 consecutive days, 24h prior to the endometriosis model was established. The label-retaining cells(LRCs) were identified by getting BrdU labeling rate by immunohistochemistry on 4h,1w, 3w, 5w, 6w,7w,8w,9w when the last injection was finished. The expression level of CD34 and CD146 on LRCs were detected by double staining of immunohistochemistry. Result  The maxium BrdU labeling rate was 69.89±0.78% and 73.27±1.06% for epithelial and stromal respectively. After a 6-week and an 8-week chase, 2.07±0.19% epithelial cells and 4.63±0.72% stromal cells retaind BrdU labeling respectively, which were considered to be epithelial and stromal LRCs.Both the epithelial LRCs and the stromal LRCs were mainly located in the boundary of the lesions and the mice tissues where the lesion was implanted. The positive expression rate of CD34 and CD146 were 19.32±0.03% and 14.76±0.03% respectively. CD34 did not co-expressd on LRCs, CD146 co-expressed on a few stromal LRCs. Conclusion  Two kinds of stem cells were identified successfully in ectopic endometrium by label-retaining cell approach, which were epithelial and stromal LRCs located in the boundary of the lesions and the mice tissues where the lesion was implanted. It seems that CD34 was not the marker for ectopic endometrial stem cells, whether CD146 can be the candidate marker for ectopic endometrial stem cells or not by combining with other markers deserving further research.

Key words: Endometriosis, stem cell, staining and labeling, bromodeoxyuridine, immunohistochemistry, antigens, CD34, 抗原