The Establishment And Evaluation Of Visualization In Nude Mouse Endometriosis Model

Objective:This study was based on the characteristics of endometriosis, and collected the eutopic and ectopic endometrium cells from patients with chocolate cyst of ovary for in vitro culture, after that, the cultured cells were injected subcutaneously into the nude mice to establish animal models of endometriosis and evaluated them then, which may provide a reliable model for further research on pathogenesis of endometriosis.Method:First, the eutopic and ectopic endometrium cells were selected from patients with chocolate cyst of ovary and cultured in vitro, which were divided into separate groups (glandular epithelial cells group, stromal cells group and mixed group), then the growth characteristics of cells was observed, Giemsa and immunofluorescence staining were used for cell morphological and specific identification (expression of vimentin and keratin), at last, the different amount (1.0×106、2.5×106、5.0×106、1.0×107) of cell suspension in each group were injected subcutaneously into the nude mice (5-6w), then the changes of injection sites at different stages were observed, at the same time, the injection sites were dissected to produce lesion specimens for HE staining (observe the pathological histology at different stages by use light microscopy), then for further clarifying the tissue cell types, the ectopic lesions which had the glandular growth were detected by immunohistochemistry, at last, the time and rate of being model by different methods were compared and analysed.Results:The epithelial cells could not be subcultured in vitro, while the stromal cells and mixed cells in each group could be subcultured, and the mixed cells cultured had a stronger proliferation than stromal cells, the difference was statistically significance (P<0.05), in the mixed cultured cells, the ectopic cells had a stronger proliferation than the eutopic cells, the difference was statistically significant (P<0.05). In the course of the experiment, good growth of ectopic lesions could be perceived by the unaided eye, which showing for the uplift of the vesicle-like structures, surface with angiogenesis and connective tissue covering, and the pathology confirmed that there were endometrial glands, glandular epithelial and stromal cells growth at the cell transplantation sites; at the first5.days, the ectopic lesion volume formed in each group was different, the difference was statistically significant (P<0.05), after10and15days, the difference was not statistically significant (P>0.05). In the eutopic group, different amouts of cells resulted different mold rates at5,10and15days, the difference was statistically significant (P<0.05), and there was no difference between5.0×106and1.O×107(P>0.05); In the ectopic group, different amouts of cells resulted different mold rates at5,10and15days, the difference was statistically significant (P<0.05), and there was no difference between5.0×006and1.0×107(P>0.05); when the number of cells were5.0×106, the mold rate of ectopic group was higher than that of eutopic, and the difference was statistically significant (P<0.05)Conclusion:The glandular epithelial cells could be subcultured in vitro when it cultured with stromal cell, the stromal cells could be subcultured, but it had a lower proliferation than mixed cells, the ectopic cells had a stronger proliferation than the eutopic cells in the mixed cultured cells; The ectopic endometrial cells (5-6generation) in mixed cultured could successfully build a mold of endometriosis when they were injected subcutaneously into the abdomen of female nude at the concentration of5.0×106/200ul, and the method could provide a visualization mold at a higher molding rate and shorter time.