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대한생식의학회지 제35권 제1호 2010년
Progesterone과 TGF-β1에 의해 탈락막화가 유도된 인간 자궁내막세포의 삼차원 공배양이 2-세포기 생쥐배아의 체외발달에 미치는 영향
부산대학교 의학전문대학원 산부인과학교실1, 부산대학교병원 불임클리닉2
권욱현1•김휘곤1•이동형1•고경래2•이규섭1*,
Effects of 3-dimensional Co-culture of Human Endometrial Cells Decidualized with Progesterone and TGF-β1 on the Development of Mouse 2-cell Embryos In Vitro
Wook-Hyun Kwon1, Hwi-Gon Kim1, Dong-Hyung Lee1, Kyung-Rae Ko2, Kyu-Sup Lee1*
1Department of Obstetrics and Gynecology, School of Medicine, Pusan National University, Busan, Korea,2Infertility Clinic, Pusan National University Hospital, Busan, Korea
Objective: This study was carried out to investigate the effects of 3-dimensional co-culture of human endometrial cells decidualized with progesterone and TGF-β1 on the development of 2-cell mouse embryos. Methods: Stromal and epithelial cells isolated from human endometrial tissue were immunostained for cytokeratin and vimentin. Expression of TGF-β1, its receptor-1,-2, integrin-β3 and prolactin in mono or co-culture according to three different hormone conditions was investigated by RT-PCR. Differential staining was used to investigate the number of ICM and trophectoderm of hatched mouse blastocysts in different three conditions. Results: The immunohistochemical study was positive for cytokeratin or vimentin and confirmed that epithelial and stromal cells were isolated from endometrial tissue successfully. In co-culture, TGF-β1, its receptor-1, integrin-β3 and prolactin except TGF-β1-r2 were expressed in progesterone dominant condition. The hatching and attaching rate were higher in the co-culture with decidualized cells (p<0.05). However, we observed that lots of the incomplete hatched blactocysts attached on non-decidualized cells. The ICM number of hatched mouse blastocysts was higher in co-culture with decidualized and non decidualized cells than media only culture (p<0.05). The trophectoderm number of hatched blastocyst was higher in the co-culture with decidualized cells than non-decidualized cells or media only culture (p<0.05). Conclusion: The administration of progesterone, estrogen and TGF-β could induce decidualization of stromal and epithelial cells isolated from human endometrial tissue using 3-dimensional co-culture, and the decidualization of human endometrial cells could increase the hatching and attaching rate of 2-cell mouse embryos.
키워드 : Decidualization, TGF-β1, Co-culture, Progesterone, RT-PCR
교신저자 : kuslee@pusan.ac.kr
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