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대한생식의학회지 제25권 제1호 2010년
동결보존시 생쥐전핵배아의 시기에 따른 생존율과 발생율의 비교
서울대학교 의학연구원 인구의학연구소, 서울대학교 의과대학 산부인과학교실
김희선, 류범용, 오선경, 서창석, 김석현, 최영민, 김정구, 문신용, 이진용,
Effects of Pronuclear Age for Freezing in Mouse Embryos Survival and Development in Vitro after Cryopreservation
Kim hs, Ryu by, Oh sk, Suh cs, Kim sh, Choi ym, Kim jg, Moon sy, Lee jy
This study was designed to evaluate the influence of pronuclear age on the survival and post-thawing development after cryopreservation of mouse embryos. Freezing and thawing were performed in the different pronuclear stages of mouse embryos after IVF. Embryos were obtained from F1 hybrid mice and classified into 4 groups according to the pronuclear stage ( 6hr, 9hr, 12hr and 15hr after insemination). Pronuclear ova were slowly cooled in a biological freezer using 1.5M 1,2-propanediol and 0.1M sucrose as cryoprotectant. Thawing was done at room temperature and 1,2-propanediol was removed by multi-step dilutions. Both frozen-thawed embryos and control fresh embryos were cultured in vitro in Ham's F-10 medium supplemented with 4mg/ml BSA. In control group, the development rate after 48hr was 99.3%, and the complete hatching rate after 144hr was 61.3%. In experimental groups, the survival rate after thawing was 95.4% in 6hr, 88.7% in 9hr, 75.2% in 12hr and 62.4% in 15hr after insemination, the development rate after 48hr was 61.1, 77.0, 67.0 and 79.6%, respectively, and the complete hatching rate after 144hr was 25.7, 43.7, 42.2 and 60.0%, respectively. The survival rate in 15hr was significantly lower ( p<0.05) compared with other groups. In vitro development rates after 48hr were similar in all groups, but complement hatching rate was significantly lower ( p<0.05 ) in 6hr group. In conclusion, cryopreservation of mouse pronuclear ova with 2 distinct pronuclei ( 9hr and 12hr groups) showed better results after thawing compared with early (6hr group) or late pronuclear ova just prior to cleavage (15hr group).
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