人子宫内膜间充质干细胞的分离、培养与鉴定

发布时间：2018-12-25 12:12

【摘要】：目的建立一种体外分离、纯化、培养和扩增子宫内膜间充质干细胞(MSCs)的方法,并对培养的子宫内膜MSCs进行鉴定。方法采用机械和胶原酶消化相结合的方法分离、纯化子宫内膜间质细胞,再利用克隆纯化法分离子宫内膜MSCs。对得到的子宫内膜MSCs进行传代培养和扩增。流式细胞术检测细胞表面抗原CD90、CD105、CD146、CD34和CD45的表达。将第3代子宫内膜MSCs冻存1个月后再复苏,鉴定复苏后细胞的免疫表型。结果子宫内膜间质细胞可形成克隆,从大克隆可分离出MSCs。流式细胞术检测得到的子宫内膜MSCs表达MSCs的标志物CD90、CD105和CD146,不表达造血干细胞的表面标志物CD34和CD45。冻存再复苏后的细胞活力达95%以上,细胞免疫表型与冻存前相似。结论该方法可以较好地分离子宫内膜MSCs,是一种实用、可靠、高效的分离方法。
[Abstract]:Objective to establish a method for isolation, purification, culture and amplification of endometrial mesenchymal stem cell (MSCs) in vitro, and to identify the cultured endometrial MSCs. Methods the endometrial stromal cells were purified by mechanical and collagenase digestion, and the endometrial MSCs. was isolated by cloning and purification. The endometrial MSCs was subcultured and amplified. The expression of CD90,CD105,CD146,CD34 and CD45 was detected by flow cytometry. The third generation of endometrial MSCs was cryopreserved for 1 month before resuscitation to identify the immunophenotype of the cells after resuscitation. Results MSCs. could be isolated from endometrial stromal cells by cloning. Flow cytometry was used to detect the expression of MSCs in endometrial MSCs, CD90,CD105 and CD146, did not express the surface markers CD34 and CD45. of hematopoietic stem cells. After cryopreservation and resuscitation, the cell viability was over 95%, and the immunophenotype of the cells was similar to that before cryopreservation. Conclusion this method is a practical, reliable and efficient method for the isolation of endometrial MSCs,.
【作者单位】：中国中医科学院江苏分院;江苏省中医药研究院妇产科;
【基金】：江苏省自然科学基金(BK20131457)
【分类号】：R329.2

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