酶消化结合差时贴壁的方法体外分离培养人子宫内膜细胞  

Isolation and cultivation of human endometrial cells in vitro with enzymolysis and differential adhesion method

作　　者：刘明慧[1] 晁贺[1] 张亚兰[2] 王鹏[2] 李建梅[3] 王淑玲[4] 刘英[1] 

机构地区：[1]首都医科大学附属北京妇产医院生殖中心,100026 [2]首都医科大学附属北京妇产医院妇科,100026 [3]首都医科大学附属北京妇产医院手术室,100026 [4]北京大学干细胞研究中心,100083

出　　处：《中国优生与遗传杂志》2013年第10期113-115,F0002,共4页Chinese Journal of Birth Health & Heredity

基　　金：北京市自然科学基金项目(5122015);北京市科技新星基金项目(H020821200190);首都医科大学附属北京妇产医院院内基金项目(201114)

摘　　要：目的探讨酶消化结合差时贴壁培养的方法对人子宫内膜细胞进行体外培养的培养效果,以寻找一种简单高效的人子宫内膜细胞体外分离培养的方法。方法采用胶原酶消化结合差时贴壁的方法,对人正常子宫内膜细胞进行体外分离培养,并传代、冻存及复苏。光镜下观察其培养效果,通过免疫荧光法对腺上皮细胞及间质细胞进行鉴定,并分析其纯度。结果共培养人子宫内膜38份,成功培养35份,培养成功率92%,冻存后成功复苏率达97%;子宫内膜腺上皮细胞和间质细胞分别经角蛋白单抗和波形蛋白单抗免疫荧光显色为阳性,腺上皮细胞和间质细胞原代纯度均达90%以上。结论酶消化结合差时贴壁的培养方法,操作简单、培养成功率高、污染机会少、省时、维持细胞活性好,是子宫内膜细胞体外分离培养的简单高效方法。Objective： To investigate the effectiveness of culturing human endometrial cells by enzymolysis and differential adhesion method, looking for a simple and efficient method for the isolation and culture of human endometrial cells in vitro. Methods ： Human endometrial cells were isolated and cultured by enzymolysis and differential adhesion in vitro. The endometrial cells were then observed by a light microscopy and verified by immunofluorescence. Results ： The endometrial cell culture was successful in 35 of 38 endometrium samples. The success rate is 92%. The cryopreservation recovery rate is 97%. Endometrial glandular epithelial cells and stromal cells were positive for keratin monoclonal antibody and vimentin monoclonal antibody staining, respectively. The purities of glandular epithelial cells and stromal ceils were more than 90%. Conclusion： Human endometrial cells were successfully isolated and cultured by enzymolysis and differential adhesion in vitro. This method is simple with high success rate, less chance of contamination, saving time, and is a simple and efficient way to isolate and culture endometrial cells in vitro.

关 键 词：子宫内膜 细胞培养 差时培养法 

分 类 号：R329-33[医药卫生—人体解剖和组织胚胎学]