[1]郭玉琪,赵潇丹,路平,等.体外分离子宫内膜干细胞3种方法的比较*[J].郑州大学学报(医学版),2012,(01):20.
 GUO Yuqi,ZHAO Xiaodan,LU Ping,et al.Comparison of three methods isolating endometrial stem cells in vitro[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2012,(01):20.
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体外分离子宫内膜干细胞3种方法的比较*
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2012年01期
页码:
20
栏目:
论著
出版日期:
2012-01-20

文章信息/Info

Title:
Comparison of three methods isolating endometrial stem cells in vitro
作者:
郭玉琪赵潇丹路平张展#
郑州大学第三附属医院妇产科 郑州 450052
Author(s):
GUO YuqiZHAO XiaodanLU PingZHANG Zhan
Department of Gynecology and Obstetrics,the Third Affiliated Hospital,Zhengzhou University,Zhengzhou 450052
关键词:
子宫内膜干细胞细胞分离
Keywords:
human endometrial stem cellcell separationhuman
分类号:
R711
摘要:
目的:探索简便、实用、高效的体外分离、培养人子宫内膜干细胞的方法。方法:采用胰蛋白酶、胶原酶Ⅰ和混合法3种体外细胞分离方法分离子宫内膜干细胞,并采用相同的方法进行培养。倒置显微镜下观察细胞消化、贴壁时间,纯化情况和细胞生长形态。绘制第2代细胞生长曲线。采用免疫细胞化学法检测第2代细胞CD90和CD146蛋白的表达,采用RTPCR法检测CD90和CD146 mRNA的表达。结果:胶原酶Ⅰ法分离子宫内膜干细胞具有操作简便,获得细胞纯度高,细胞易于贴壁,生长良好的优势。胶原酶Ⅰ法分离的细胞经培养后获得的细胞符合正常人子宫内膜干细胞形态学特征和生物学特征;第2代细胞生长曲线均为近似的“S”形;第2代细胞CD90和CD146蛋白的表达为阳性,CD90和CD146 mRNA有表达。结论:胶原酶Ⅰ法较为简便、实用并具有较高的成功率,是一种培养子宫内膜干细胞的可行性方法。
Abstract:
Aim: To explore a simple, practical and efficient approach of isolation human endometrial stem cells in vitro.Methods:Trypsin, collagenaseⅠ,and mixed ways were used to isolate human endometrial stem cells,respectively.Immunocytochemistry and RTPCR method were used to identify expressions of CD90 and CD146 in the endometrial stem cells.Results:By comparison,the collagenase Ⅰ was the simplest way to obtain high purity, easyadherent cells.The cells were consistent with normal morphological characteristics and biological characteristics of endometrial stem cells by the collagenase Ⅰ method.Secondgeneration cell growth curves were similar to “S” shape.The expressions of CD90 and CD146 were positive,and the CD90 and CD146 mRNA expressed.Conclusion:The collagenaseⅠ digestion method of isolating endometrial stem cells is simple, practical and has a high success rate,which is a feasible separation method of human endometrial stem cells.

参考文献/References:

[1]Meng X,Ichim TE,Zhong J,et al.Endometrial regenerative cells:a novel stem cell population[J]. J Transl Med,2007,5:7A [2]Musina RA,Belyavski AV,Tarusova OV.Endometrial mesenchymal stem cells isolated from the menstrual[J].Blood,2008,45(4):539 [3]Li L, Xie T. Stem cell niche:structure and function[J].Annu Rev Cell Dev Biol,2005,21(3):605 [4]习佳飞,王韫芳,裴雪涛. 成体干细胞及其在再生医学中的应用[J]. 生命科学,2006, 8(4):328 [5]Meng X,Ichim TE,Zhong J,et al. Endometrial regenerative cells:a novel stem cell population[J].J Trans Med,2007,15(5):57 [6]Schwab KE,Gargett CE.Coexpression of two perivascular cell markers isolates mesenchymal stemlike cells from human endometrium[J].Hum Reprod,2007,22(11):2903

备注/Memo

备注/Memo:
*郑州市科技局科技计划基金资助项目112PPPF3189 doi:10.3969/j.issn.16716825.2012.01.007 #通讯作者,女,1959年9月生,主任技师,教授,研究方向:生殖免疫,Email:Zhangzhan@yahoo.com.cn
更新日期/Last Update: 1900-01-01