种植窗期子宫内膜细胞体外培养的形态学特点与妊娠结局的关系

The correlation between pregnancy outcome and morphological characters of endometrial cells culture in vitro

目的对种植窗期人子宫内膜进行体外培养，探讨不同妊娠结局的子宫内膜细胞体外培养的细胞形态学特点。方法收集拟行IVF-ET助孕的患者种植窗口期（排卵后第5～7天）子宫内膜组织进行体外培养，于倒置显微镜下观察子宫内膜细胞形态学特性，并通过免疫荧光法进行鉴定，采用培养板单层贴壁细胞的原位计数方法，计算子宫内膜间质细胞和腺上皮细胞数量及比例。按助孕后妊娠结局，分为妊娠组和未妊娠组，比较两组患者子宫内膜细胞体外培养的细胞形态学特点。结果39例子宫内膜组织，培养成功36例。其中妊娠组19例，培养成功18例，培养成功率94.7％；未妊娠组20例，培养成功18例，培养成功率90.0％，两者的培养成功率差异无统计学意义（P〉0.05）。子宫内膜间质细胞接种后即开始贴壁，细胞呈扁平状，胞浆透明，核圆居中，波形蛋白表达阳性。子宫内膜腺细胞于接种2h后开始贴壁，细胞呈多角形或蝌蚪形，旋涡状排列，核圆而大，角蛋白表达阳性。妊娠组腺细胞贴壁率为64.8％±4.7，贴壁培养后腺上皮细胞的比例为17.3％±9.5，间质细胞比例为82.7％±9.5；未妊娠组子宫内膜组织腺细胞贴壁率为57.6％±3.4，贴壁培养后腺上皮细胞的比例为14.7％±4.8，间质细胞比例为85.3％±4.8。妊娠组比未妊娠组腺上皮细胞贴壁率及贴壁后腺细胞比例高，但差异无统计学意义（P〉0.05）。结论种植窗期人子宫内膜细胞进行体外培养后，妊娠组与未妊娠组细胞的外观特征相似，虽妊娠组腺上皮细胞贴壁率及贴壁后腺细胞比例较高，但两组差异无统计学意义。单以种植窗期子宫内膜体外培养的细胞形态特点，无法判断内膜的容受性，也无法预测妊娠结局。

Objective： To investigate the morphological characteristics of endometdal cells during implantation window with different pregnancy outcomes. Methods： A small mount of endometrial tissue was collected for in-vitro cultivation from patients scheduled for IVF-ET during their implantation window （day 5～7 after ovulation） . The Endometrial cells were observed by a light microscopy and verified by immunofluorescence. By counting cell numbers under microscopy randomly, we calculated the endometrial stromal cells and glandular epithelial cells proportion. Results： 1.36 out of 39 endometrial specimens biopsied during implantation window were cultured successfully in-vitro, the resting 3 cases failed because of the too small amount of endometrial volume when biopsied. 18 out of 19 cases in pregnant group were cultured successfully and the success rate was 94.7%. 18 out of 20 cases in non-pregnant group were successful and the rate was 90.0%. The successful culture rate were not statistically significant between two groups （P〉0.05） .2. Endometrial stromal cells adhered to the plate right after seeding. They were flat, transparent cytoplasm, nuclear circle center and positive for vimentin monoclonal antibody. Glandular epithelial cells began attached to the plate 2 hours after seeding. They were polygonal or tadpole-shaped cells, with vortex-like arrangement, the nucler were round and large and positive for keratin monoclonal antibody staining.. 3. In the pregnant group, the adherent rate of primary endometrial glandular cells was 64.8% ± 4.7. The proportion of glandular cells was 17.3% ± 9.5 while the stromal cells was 82.7% ±9.5 of all the attached cells. In the non-pregnant group, the adherent rate of primary endometdal glandular cells was57.6% ± 3.4, the proportion of glandular cells was 14.7% ± 4.8, while the stromal cells was 85.3% ±4.8. 4. The adherence rate of glandular cells was less than stromal cells in non-pregnant group, but the differences were not statistically significant （P〉0.05） . Conclusions： Human endometrial cells cultured in vitro shared similar morphological characteristics between pregnant and non-pregnant groups. Although the epithelial cell adhesion rate and the proportion of glandular cells in attached cells are higher in pregnant group than non-pregnant group, the difference is not statistically significant, which offers a platform for researching endometrium functions at cellular and molecular levels.