In VitroDecidualization and Molecular Characterization of Murine Endometrial Stromal Cells.

초록

In VitroDecidualization and Molecular Characterization of Murine Endometrial Stromal Cells. Jungwoo Kim,1 Yoon Young Kim, 1 Yong Jin Kim,2 Sung Woo Kim,1 Hoon Kim,1 Seung-Yup Ku.1 1 Seoul National University Hospital, Seoul, Korea, Republic of; 2 Korea University Guro Hospital, Seoul, Korea, Republic of. Introduction: In pregnancy, the decidualization of uterine endometrial cells has a pivotal role in the implantation of a fertilized embryo. Decidualization of endometrium includes drastic functional and morphological changes of endometrial stromal cells and can lead to the establishment of a successful pregnancy. Endometrial decidualization is caused by a complex combination of transcription factors, morphogens, cytokines, cell cycle regulators, and signaling pathways. However, the deeper knowledge of molecular characteristics and mechanisms of reception of embryo by decidualized endometrial cells were not sufciently elucidated. Methods: Uterine horns of 6 weeks old female C57BL/6 mice were isolated and the endometrial stromal cells were extracted using collagenase type I (1 mg/ml). Collected cells were expanded and treated under each condition, 1) P4, 2) P4+E2, 3) P4+ cAMP, 4) cAMP, for 14 days. The expression of specifc genes and proteins were analyzed using RT-qPCR and immunoblotting. Results: Here, we were focusing on finding the best condition of inducing decidualization in mouse endometrial stromal cells. We found that three important molecules are mainly involved in the induction of decidualization, such as cyclic adenosine monophosphate (cAMP), estradiol (E2), and progesterone (P4). We confrmed that the groups of treatment with P4 and P4+E2 for 14 days show morphological change like decidualization, which are representatively contracted cytosol. Also, we found that forkhead box O1 (FOXO1), which is a well-known transcription factor in the decidualization process, and desmin, which is an intermediate flament expressed for decidualization, are highly expressed in decidualized cells. Most importantly, the expression of a decidual marker, such as decidual prolactin (PRL), was confrmed by western blot analysis. These results indicate that P4 and P4+E2 treatment can stimulate endometrial stromal cells to transform into a decidualization state. Conclusion: These results indicate that our experimental system generates in vitro decidualized mouse endometrial cells and can reveal signals or factors increasing proper communications between the embryo and decidualized endometrial cells, even we address the potential methods to cure infertility because of abnormality of endometrial reception and communication network between them (2020R1A2C1010293 and 2020R1F1A1076286).