关于改良差速贴壁法原代培养大型成年哺乳动物绵羊肌源性干细胞的研究  

A study on isolation and purification adult sheep muscle-derived stem cells by modified preplate technique

作　　者：李玢[1] 邓卓 甘露[1] 姜向阳[1] 张璐[1] 

机构地区：[1]陕西省人民医院妇科,陕西西安710068

出　　处：《中国计划生育和妇产科》2017年第12期25-29,共5页Chinese Journal of Family Planning & Gynecotokology

基　　金：国家自然科学基金青年项目(项目编号:81501238)

摘　　要：目的探讨改良差速贴壁法原代培养成年大型哺乳类动物绵羊肌源性干细胞(muscle-derived stem cells,MDSC)的可行性。方法改良差速贴壁法分离培养原代绵羊MDSC,并通过免疫荧光检测及western blotting法检测干细胞抗原-1(stem cell antigen,Sca-1),Desmin及CD 34表达情况。Western blotting法检测贴壁细胞(pp)1~pp6的Desmin表达情况来反应纯化情况。结果利用改良差速贴壁法可以成功分离培养出绵羊MDSC,该干细胞Sca-1,Desmin及CD 34表达均阳性,贴壁慢的细胞Desmin表达增高,在pp6中表达最高。结论改良差速贴壁法可以成功分离培养原代绵羊MDSC,且可扩增至20代。Objective Feasibility study of modified preplate technique for isolating the sheep muscle - derived stem ceils （MDSC）. Methods Modified preplate technique for isolation the sheep MDSC was optimized and the expressions of Sca- 1, desmin and CD 34 were detected by immunofluorescenee stain and western blotting assay. The expression of desmin of ppl - pp6 was detected by western blotting assay to learn the effect of purification MDSC by modified preplate technique. Results The adult female sheep MDSC was isolated successfully using the modified preplate technique in vitro. The MDSC was positive expression of Sca - 1, desmin and CD 34. The expression trend of desmin in ppl - pp6 was related to the speed of attachment. The slower the speed, the higher the desmin expressed. The expression of desmin was highest in pp6. Conclusion Modified preplate technique can isolate the adult female sheep MDSC successfully. It has a self - renewal characteristics of stem cells that can be amplified more than 20 generations.

关 键 词：肌源性干细胞 盆底功能障碍性疾病 组织工程 再生医学 

分 类 号：R711[医药卫生—妇产科学]