| Establishment of Three Dimensional in-vitro Culture System with Human Endometrial Cells: Induction of Differentiation by Sex Steroid Hormone & Characterization. |
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Hyuk Chan Gwon, Won Il Park, Sun Young Kim, Dong Wook Park, Mi Ran Kim, Sung Won Han, Dae Woon Kim, Ho Joon Lee, Su Yoo Hong, Joon Young Park, Se Gwang Kim, Dong Je Jo |
1Department of Obstetrics and Gynecology, Eulji University School of Medicine, Seoul, Korea.
2Department of Molecular Science and Technology, Graduate School, Aj ou University, Suwon, Korea.
3Department of Obstetrics and Gynecology, Aj ou University School of Medicine, Suwon, Korea.
4Department of Physiology, Eulji University School of Medicine, Seoul, Korea.
5Department of Obstetrics and Gynecology, Yonsei University College of Medicine, Seoul, Korea. |
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| Abstract |
OBJECTIVE
The aim of this study was to establish three-dimensionally cultured endometrial cell model containing endometrial stromal cell (ESC), endometrial epithelial cell (EEC) and extracellular matrix (ECM) and to compare the morphological and biomolecular expression patterns of this model with mid-luteal endometrium in vivo. MATERIALS AND METHODS: The EEC and ESC was obtained from hysterectomy specimen and cultured separately. The EEC was overlayered in Matrigel layer on ESC embedded in collagen. The model had been cultured for 48 h in DMEM medium containing estrogen and progesterone. The ultrastructure was evaluated by electron microscopy. The expression of integrins, cyclooxygenases and matrix metalloproteinases were examined by immunohistochemistry and zymography. RESULTS: EEC in three-dimensional culture model grew with polarity and tight junction and desmosome between cells were found. The formation of pinopodes was also detected. In three-dimensionally cultured endometrial cell model, the expression of integrin alpha1, alpha4, beta3, MMP-1, -2, -3 and 9 was detected which was not expressed in monolayer culture of EEC, ESC or ESC embedded in collagen. CONCLUSION: The three-dimensionally cultured endometrial cell model possessed the morphological and biomolecular characteristics of in vivo endometrium of implantation period. These characteristics could be achieved by paracrine interactions between ESC and EEC. This model may contribute to the studies of differentiation of endometrium, process of implantation and pathophysiology of implantation-related diseases. |
| Key Words:
Three dimensional co-culture, Human endometrial cell, Integrin, MMPs |
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