1999 Fiscal Year Final Research Report Summary
BASIC STUDIES FOR IMPROVEMENT OF IN VITRO FERTILIZATION AND CRYOPRESERVATION
| Project/Area Number |
10671518
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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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 | UNIVERSITY OF TOKYO |
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Principal Investigator |
TSUTSUMI Osamu UNIVERSITY OF TOKYO, DEPARTMENT OF OBSTETRICS AND GYNECOLOGY, PROFESSOR, 医学部・附属病院・分院, 教授 (60134574)
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| Project Period (FY) |
1998 – 1999
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| Keywords | embryo / cryopreservation / in vitro fertilization / glucose / vitirification / GLUT / implantation |
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| Research Abstract |
Effects of two cryopreservation procedures (conventional slow controlled-rate freezing using a programmable freezer and vitrification by direct plunging into liquid nitrogen) were compared on 2-cell embryos and their subsequent development to blastocysts, fresh or cryopreserved 2-cell mouse embryos were developed into blastocysts in vitro. The percentage of vitrified embryos which developed into blastocysts was significantly lower than that of fresh and slow controlled-rate frozen embryos. Although blastocysts from each cryopreservation procedure appeared morphologically normal and neither number of cells in the blastocysts nor in-vitro trophoblast spreading differed significantly, there were significant differences in their functional viability. First, the glucose incorporation activity in terms of [(3)H]2-deoxyglucose (2-DG) uptake in vitrified and thawed 2- cell embryos significantly decreased compared with fresh or slow controlled-rate frozen and thawed 2-cell embryos. Second, 2-DG uptake by blastocysts developed in vitro from fresh 2-cell embryos and from slow controlled-rate frozen or vitrified 2-cell embryos was 105 +/- 75, 43.0 +/- 28.3 and 22.O +/- 11.4 fmol/embryo/h respectively. Third, the implantation rate of blastocysts developed in vitro from vitrified 2-cel1 embryos (10.2%) was significantly lower than that from fresh 2-cell embryos (30.8%) or slow controlled-rate frozen 2-cell embryos (22.1%). Since these data suggest that cryopreservation may have ulterior consequences on the functional development of embryos and that vitrification may exert a more harmful effect than slow controlled-rate freezing, more attention should be paid to its safety before vitrification is used routinely in a clinical programme.
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[Publications] Osuga, J., Ishibashi, S., Oka. T., Yagyu, H., Tozawa, R., Fujimoto, A.. Shionoiri. F., Yahagi, N., Kraemer, F. B., Tsutsumi, O. and Yamada, N.: "Targeted disruption of hormone-sensitive lipase results in male sterility and adipocyte hypertrophy, but riot in obesity [In Process Citation]."Proc Natl Acad Sci U S A. 97(2). 787-792 (2000)
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[Publications] Kamei, Y., Tsutsumi, O., Yamakawa, A., Oka, Y., Taketani, Y. and Imaki, J.: "Maternal epidermal growth factor deficiency causes fetal hypoglycemia and intrauterine growth retardation in mice : possible involvement of placental glucose transporter GLUT3 expression."Endocrinology. 140(9). 4236-4243 (1999)
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[Publications] Hiroi, H., Momoeda, M., Inoue, S., Tsuchiya, F., Matsumi, H., Tsutsumi, O., Muramatsu, M. and Taketani, Y.: "Stage-specific expression of estrogen receptor subtypes and estrogen responsive finger protein in preimplantational mouse embryos."Endocrine Journal. 46. 153-158 (1999)
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[Publications] Osuga, Y., Tsutsumi, O., Momoeda, M., Okagaki, R., Matsumi, H., Hiroi, H., Suenaga, A., Yano, T. and Taketani, Y.: "Evidence for the presence of hepatocyte growth factor expression in human ovarian follicles."Molecular Humman Reproduction. 5. 703-707 (1999)
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[Publications] Tsutsumi, O., Uechi, H., Sone, H., Yonemoto, J., Takai, Y., Moeda, M., Tohyama, C., Hashimoto, S., Morita, M. and Taketani, Y.: "Presence of dioxins in human follicular fluid: their possible stage- specific action on the development of preimplantation mouse embryos."Biochemical Biophysical Research. 250. 498-501 (1998)
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