Mulholland J, Winterhager E, Beier H M
Department of Anatomy and Reproductive Biology, RWTH University of Aachen, Federal Republic of Germany.
Cell Tissue Res. 1988 Apr;252(1):123-32. doi: 10.1007/BF00213833.
Morphological and biochemical changes occurring in rabbit endometrial epithelial cells when placed in culture were investigated. Cells were examined by scanning- and transmission electron microscopy and freeze-fracture. Morphologically, cultured cells are shorter and broader than the columnar epithelial cells in vivo, but retain their polarity as indicated by the presence of apical microvilli and a well-developed junctional belt. To study changes in biochemical function, proteins synthesized by cells in primary culture were analyzed by two-dimensional gel electrophoresis. Proteins were labeled during a 24-h incubation with 35S-methionine and gels examined by fluorography. The pattern of proteins changed after cells had been in culture for 48 h. On day 3 new proteins were synthesized and several protein species labeled during days 1 or 2 of culture, including uteroglobin, no longer appeared. On days 3-8 of culture the protein patterns were similar. Addition of progesterone, estradiol, prolactin, or combinations of these hormones to the culture medium for 24-144 h failed to elicit consistent changes in the pattern of labeled proteins established after 3 days of culture. Minor differences in protein patterns among unrelated cultures appear to have been derived from the original cells of the culture. These results indicate that after 48 h in primary culture, cells grown in vitro resemble endometrial epithelial cells morphologically, but no longer reflect functionally the character of epithelial cells in the uterus.
研究了兔子宫内膜上皮细胞在培养时发生的形态学和生化变化。通过扫描电子显微镜、透射电子显微镜和冷冻蚀刻技术对细胞进行检查。在形态学上，培养的细胞比体内的柱状上皮细胞更短更宽，但通过顶端微绒毛和发育良好的连接带的存在表明它们保持了极性。为了研究生化功能的变化，通过二维凝胶电泳分析原代培养细胞合成的蛋白质。在24小时的孵育过程中用35S-甲硫氨酸标记蛋白质，并用荧光自显影检查凝胶。细胞培养48小时后蛋白质模式发生了变化。在第3天合成了新的蛋白质，并且在培养第1天或第2天标记的几种蛋白质，包括子宫珠蛋白，不再出现。在培养的第3 - 8天，蛋白质模式相似。将孕酮、雌二醇、催乳素或这些激素的组合添加到培养基中24 - 144小时，未能引起培养3天后建立的标记蛋白质模式的一致变化。无关培养物之间蛋白质模式的微小差异似乎源于培养物的原始细胞。这些结果表明，原代培养48小时后，体外生长的细胞在形态上类似于子宫内膜上皮细胞，但在功能上不再反映子宫上皮细胞的特征。
