论文部分内容阅读
目的 为进一步实验需要，建立子宫内膜间质细胞模型。方法 胶原酶消化联合应用差速离心和密度梯度离心以分离纯化目的细胞; 恒温孵箱培养细胞; 倒置显微镜观察细胞形态; 台盼兰染色计数细胞存活率; 免疫组化鉴定细胞。结果 该方法分离纯化子宫内膜间质细胞的存活率高，形态与子宫内膜腺上皮细胞易区别。体外培养的间质细胞具有一定分裂增殖能力，传代培养的形态特征与原代细胞一致。免疫组化显示Vimentin(+) Keratin(-)是鉴定间质细胞的标志。 结论 该方法分离子宫内膜间质细胞高效 、简便 、省时.
Objective To establish a model of endometrial stromal cells for further experimental purposes. Methods Collagenase digestion was performed by differential centrifugation and density gradient centrifugation to separate and purify the cells of interest. Cells were cultured in incubator under constant temperature. Cell morphology was observed under inverted microscope. Cell viability was counted by trypan blue staining and cells were identified by immunohistochemistry. Results The method of isolation and purification of endometrial stromal cells with high survival rate, morphology and endometrial glandular epithelial cells easily distinguishable. In vitro cultured mesenchymal cells have a certain ability to divide and proliferate, and the morphological characteristics of subculture are consistent with those of primary cells. Immunohistochemistry showed that Vimentin (+) Keratin (-) is a hallmark of stromal cells. Conclusion The method of isolation of endometrial stromal cells is efficient, simple and time-saving.