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Hyun Won Yang1 , Dong Wook Park1 , Hyuck Chan Kwon1 , Kyung Joo Hwang1 , Sei Kwang Kim2 , Dong Jea Cho2 , Kie Suk Oh1
Department of Obstetrics & Gynecology, Ajou University School of Medicine, Suwon, Korea 1
Department of Obstetrics & Gynecology, Yonsei University, College of Medicine, Seoul, Korea 2
Introduction: In order to study the human implantation mechanism, various methods for the culture of endometrial cells in vitro have been attempted. However, a disadvantage is that primary cultures of stromal and epithelial cells do not have the ability to differentiate, and therefore cannot be reproduced in the same manner as in vivo endometrium. The object of this study is to establish a 3 dimensional culture of endometrial cells which are both morphologically and functionally identical to in vivo endometrium. Materials and Methods: Endometrial tissues obtained after hysterectomies were cut into thin slices and treated with collagenase and trypsin-EDTA. The stromal cells and the epithelial cells were separated by centrifugation and cultured for 24 hours in DMEM media containing 10% FCS and 1 nM estradiol, with or without 100 nM progesterone. The cultured stromal cells were mixed with collagen gel and solidified, after which it was covered with matrigel. Epithelial cells were inoculated on the top and then cultured for 3 days. Results: The collagen gel and the 3-dimensional endometrial cell structure was maintained only when progesterone and estradiol was added. The three-dimensionally cultured endometrial cells were stained for integrin a1, 3, and cyclooxygenase-1, -2 by immunohistochemistry, all of which showed strong expression. The cultured epithelial cells showed the microvilli formation, tight junctions and pinopodes by electron microscopy. Conclusion: Progesterone and estradiol is essential for 3-dimensional endometrial cell cultures, and the histological and immunohistochemical characteristics 3 days after culture were similar to the in vivo implantation stage endometrium. It is thought that further studies into a new culture environment which would allow longer periods of culture will be necessary.
This abstract is being presented on Sunday, August 1 at 8:00 AM to 10:15 AM at CUB 2nd Floor Ballroom.