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대한생식의학회지 제29권 제3호 2010년
Cumulus Free 생쥐 성숙란의 초자화 동결-융해 후 Simple Media에서의 수정 및 배 발달
고려대학교 의과대학 산부인과학교실
정수경, 김성건,이정재,오지현,이용호,김선행
In vitro Fertilization and Embryo Development in Simple Media of the Frozen-Thawed Cumulus-free Mouse Oocytes Cryopreserved by Vitrification
Jung SK
Department of Obstetrics and Gynecology School of Medicine, Korea University
Objective: To observe the capability of fertilization and embryo development including blastocyst formation of the oocytes in simple media after thawing of the cryopreserved cumulus-free mouse oocytes by vitrification method. Methods: Oocytes were collected from 5 to 6 weeks old ICR female mice, and were denuded from the cumulus cells by 0.1% hyaluronidase. Recovered mature oocytes in study group were cryopreserved by vitrification method using EM grid for 5~7 days. In brief, oocytes were exposed in dPBS containing 1.5 M EG and 5.5 M EG+1 M sucrose for 2.5 minutes and 20 seconds each, and then executed vitrification by plunging in LN2 after loading on EM grid. Thawing treated by exposure of 1, 0.5, 0.25 and 0.125 M sucrose solution for 2.5 minutes each in order and used for experiments. Spermatozoa aspirated form the epididymis of 12 weeks old ICR male mice were used for insemination after capacitation. T6 media containing 0.4% BSA were used for fertilization and development. Results: Survival and fertilization rates after thawing were 76.9% and 79.6% respectively. Fertilization rate was lower (p<0.005) than that of control group (92.9%). There was no difference in embryo developmental rates from 2-cell to morula, however, the blastocyst formation rate and mean cell numbers of blastocysts in study group (63.3%, 58.9¡¾9.2) were lower compared with those of control group (76.1%, 63.5¡¾8.9). Conclusion: Vitrification is an effective method for mouse mature oocyte cryopreservation with high survival and fertilization rate after thawing. And in simple media, fertilization rates and embryo development of frozen-thawed mouse oocytes are satisfactory.
키워드 : : Frozen-Thawed by vitrification, Fertilization, Embryo development
교신저자 : derosi@hanmail.net
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