| Project/Area Number |
09470357
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | Shiga University of Medical Science |
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Principal Investigator |
NODA Yoichi Shiga University of Medical Science, School of Medicine, Professor, 医学部, 教授 (50115911)
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| Co-Investigator(Kenkyū-buntansha) |
TAKEBAYASHI Koichi Shiga University of Medical Science, School of Medicine, Assistant Professor, 医学部, 助手 (20303779)
KIMURA Toshio Shiga University of Medical Science, School of Medicine, Assistant Professor, 医学部, 助手 (40273411)
HIROSE Masaya Shiga University of Medical Science, School of Medicine, Lecturer, 医学部, 講師 (50238408)
TAKAKURA Kenji Shiga University of Medical Science, School of Medicine, Associate Professor, 医学部, 助教授 (10221350)
喜多 伸幸 滋賀医科大学, 医学部, 助手 (20273419)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥8,600,000 (Direct Cost: ¥8,600,000)
Fiscal Year 1998: ¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 1997: ¥4,700,000 (Direct Cost: ¥4,700,000)
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| Keywords | mouse / endometrial stromal cells / decidualization / co-culture of embryo and endometrial stromal cells / 胚共培養と子宮内膜間質細胞 / 脱落膜細胞 / マウス胚 |
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| Research Abstract |
Progress in understanding the mechanism of implantation has been hampered by the complexity of cellular interactions between embryos and endometrial stromal cells (ESCs). Although in vitro system of implantation may contribute to understanding such complex biological phenomena in vivo, it is ethically impossible to use human embryos in these experiments. Therefore, the establishment of in vitro model of mouse implantation is mandatory to understand the mechanism of implantation. As a first step, we tried to establish in vitro model of decidualization. Mouse ESCs were collected and incubated in DMEM supplemented with 10% FCS, estradiol (E2, 0.1nM) and progesterone (P, 100nM).ESCs cultured with EP transformed into large and decidua-like cells and produced decidual protein. Ultrastructurally, these cells became to have abundant rough endoplasmic reticulum during the transformation. These findings show that mouse ESCs in culture respond to ovarian steroids and showed morphological and func
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tional changes similar to decidualization in vivo. Thus, this culture system may serve as an in vitro model of mouse decidualization.
As a second step, development of embryos and invasion of the trophoblast into ESCs were studied using this in vitro. model of decidualization. ESCs isolated from 4 week old mice were cultured for 9 days in DMEM supplemented with E2 (0.1 nM) and P (100 nM). The blastocysts isolated from 4 day pregnant mice were co-cultured with ESCs with or without E2/P, and the blastocysts were also cultured without ESCs as control (single culture). After 12-day culture, the blastocysts development and their interaction with ESCs were examined under light microscopy and transmission electron microscopy. The results were as follows : 1) blastocysts in the single culture were degenerated by the 12th day of culture regardless of E2/P addition, while the blastocysts co-cultured with ESCs survived without degeneration ; 2) in the co-culture, the area of growing embryos significantly increased in E2/P containing medium, compared with those without E2/P ; 3) the invasion of trophoblasts into ESCs was apparent in medium without E2/P, while it was inhibited in medium with E2/P. ; 4)ESCs were transformed into decidua-like cells in spite of culture without E2/P when they were co-cultured with blastocysts. These results suggest that decidual cells enhance the embryonic development and regulate the trophoblastic invasion, while embryos may produce some decidualization-stimulating factor(s), thus indicating the presence of a mutual interaction between decidualized ESCs and embryos. Less
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