子宫内膜异位症体外细胞模型的建立

目的从子宫内膜组织分离腺上皮细胞和间质细胞，建立子宫内膜异位症体外多细胞模型。方法收集子宫内膜异位症和其他良性疾病的在位子宫内膜，采用高浓度胶原酶消化法进行内膜细胞的原代培养，以及传代培养。观察两组细胞形态，SABC免疫细胞化学进行细胞鉴定，MTT法绘制两组细胞的生长曲线。结果高浓度胶原酶消化后，子宫内膜异位症组和对照组腺上皮细胞和间质细胞大部分在48h内都能贴壁；腺细胞平均生长时间为6周；间质细胞平均生长时间为15周。腺上皮细胞角蛋白（cytokeratin）免疫细胞化学染色呈阳性，渡形蛋白（vimentin）为阴性；间质细胞角蛋白染色为阴性，而波形蛋白为阳性。两组生长曲线接近。结论高浓度胶原酶消化法建立子宫内膜异位症体外多细胞模型简单、易行。子宫内膜异位症在位内膜细胞的形态、生长和正常内膜细胞无明显差异。可以用作研究子宫内膜异位症细胞基因型特征、及其他因素对异位内膜细胞的影响的模型。

Establishment of endometriosis cell model in vitro

Objective To establish a multicellular model of endometriosis in vitro by separating epithelial cell and stomal cell from endometrium.Methods Eutopic endometrium of endometriosis and other benign disease was collected.Primary culture of en- dometrial cells was performed by high concentration collagenase digestion method.Morphology of endometrial cells of two groups was observed under phase-contras microscope.SABC immunocytochemistry to identify the cells,and MTT method to describe the cell growth curve of two groups were done.Results After high concentration collagenase digestion,epithelial and stromal cells of endometriosis and the control group could stick to the wall of culture flask in 48h under microscope,the average life time of epitheli- al cells was 6 weeks;stromal cells was 15 weeks.Immunocytochemical staining against cytokeratin of epithelial cell was positive, and that against vimentin was negative.On the contrary,immunocytochemical staining against vimentin of stromal cell was posi- tive,and that against cytokeratin was negative.The growth curve of two groups was close to each other.Conclusion High con- centration collagenase digestion method to establish multicellular model of endomctriosis in vitro is simple and easy.There is no distinct difference between endometrial cell of endometriosis and normal endometrium in the morphology and growth conditions.It can be used as a model to study gene characteristics of the endometrial cells and other facter's effects on them.