| Effects of 3-dimensional Co-culture of Human Endometrial Cells Decidualized with Progesterone and TGF-beta1 on the Development of Mouse 2-cell Embryos In Vitro. |
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Wook Hyun Kwon, Hwi Gon Kim, Dong Hyung Lee, Kyung Rae Ko, Kyu Sup Lee |
1Department of Obstetrics and Gynecology, School of Medicine, Pusan National University, Busan, Korea.
2Infertility Clinic, Pusan National University Hospital, Busan, Korea. |
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| Abstract |
OBJECTIVE
This study was carried out to investigate the effects of 3-dimensional co-culture of human endometrial cells decidualized with progesterone and TGF-beta1 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-beta1, its receptor-1,-2, integrin-beta3 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-beta1, its receptor-1, integrin-beta3 and prolactin except TGF-beta1-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-beta 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. |
| Key Words:
Decidualization; TGF-beta1; Co-culture; Progesterone; RT-PCR |
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