小鼠子宫平滑肌细胞的分离培养及鉴定

为寻找研究子宫平滑肌细胞生理和病理的实验研究模型,试建立一种新的简易机械分离培养小鼠子宫平滑肌细胞的方法。先采用机械分离方法分离子宫平滑肌,再用Ⅰ型胶原酶和胰蛋白酶混合消化法消化,可获得90%以上具有正常代谢活性的小鼠子宫平滑肌细胞,存活的细胞经倒置显微镜观察和免疫荧光染色进行鉴定。原代培养3d后细胞贴壁生长,约2周后细胞汇合成片,所培养的细胞在倒置显微镜下呈梭形和典型"峰-谷"样生长,平滑肌细胞肌动蛋白(α-Actin)免疫荧光染色强阳性。采用本法体外培养的小鼠子宫平滑肌细胞生长良好,纯度高,可用于小鼠子宫平滑肌细胞生理和病理等方面的研究。

Isolation Culture and Identification of Mouse Myometrial Smooth Muscle Cell

To study a new simple mechanical method for the isolation and culture of Mouse myometrial smooth cells （mMSMCs） in order to provide a experimental model for study the physiology and pathology of MSMCs. A new mechanical method was used to isolate mouse myometrial smooth. And then type Ⅰ collagenase and trypsin were used to isolate mMSMCs. More than 90%of the dissociated cells exhibited initial viability. The surviving cells were observed by phase con- trast microscope and verified by immunofluorescence staining. After 3 days of incubation, primary cultures of cells attached the wall of the incubation dishes, and reached confluence with 2 weeks. The cells grew with spindle--shape and showed typi- cal ＂hill-valley＂ pattern under phase contrast microscope. Immunofluorescence staining for smooth musclea--actin shewed positive reaction in more than 98% attached cells. The mMSMCs cultured in this experiment survive well, with highly puri- ty, and can serve the further studies of physiology and pathology.