| 학회지 검색 | HOME > 논문 및 학회지 > 학회지 검색 |
대한생식의학회지 제26권 제1호 2010년
생쥐 모델을 이용한 배아의 할구 생검법과 할구가 생검된 배아의 배양시 공배양 효과에 관한 연구 : 인간에서의 착상 전 유전진단 기술 개발을 위한 동물실험 모델의 개발
서울대학교 의과대학 산부인과학교실, 의학연구원 인구의학연구소1, 한국과학기술원 의과학연구센터2, 울산대학교 의과대학 산부인과학교실3
김석현, 류범용1, 지병철, 최성미1, 김희선1, 방명걸2, 오선경1, 서창석, 최영민, 김정구, 문신용, 이진용, 채희동3, 김정훈3,
Effects of Coculture on Development of Biopsied Mouse Embryos as a Preclinical Model for Preimplantation Genetic Diagnosis of Human Embryos
Kim sh, Ryu1 by, Jee bc, Choi1 sm, Kim1 hs, Pang2 mg, Oh1 sk, Suh cs, Choi ym, Kim jg, Moon sy, Lee
The genetic defects in human gametes and embryos can cause adverse effects on overall reproductive events. Biopsy of embryos for preimplantation genetic diagnosis(PGD) offers a new possibility of having children free of the genetic disease. In addition, advanced embryo culture method may enhance the effectiveness of embryo biopsy for the practical application of PGD. This experimental study was undertaken to evaluate the effects of coculture on the development in vitro of biopsied mouse embryos as a preclinical model for PGD of human embryos. Embryos were obtained after in vitro fertilization(IVF) from F1 hybrid mice(C57BL♀/CBA♂). Using micromanipulation, 1, 2, 3 or 4 blastomeres of 8-cell stage embryos were aspirated through a hole made in the zona pellucida by zona drilling(ZD) with acidic Tyrode's solution(ATS). After biopsy of blastomeres, embryos were cultured in vitro for 110 hours in Ham's F-10 supplemented with 0.4% BSA or cocultured on the monolayer of Vero cells in the same medium. The frequence of blastocyst formation were recorded, and the embryos beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. There was no significant difference in the blastocyst formation between the zona intact control group and the zona drilling(ZD) only, or biopsied groups. The hatching rate of all the treatment groups except 4/8 group was significantly higher than that of control group. In all the treatment groups, there was a significant reduction in the mean cell number of embryos beyond blastocyst stage(50.2±14.0 in control group vs. 41.2±7.9 in ZD, 39.3±8.8 in 7/8, 29.7±6.4 in 6/8, 25.1±5.7 in 5/8, and 22.1±4.3 in 4/8 groups, p<0.05). When the same treatments were followed by coculture with Vero cells, a similar pattern was seen in the blastocyst formation and the hatching rate. However, in all the treatment groups, there was a significant increase in the mean cell number of embryos beyond blastocyst stage with coculture, compared with the parallel groups without coculture. In the cleavage rate of biopsied blastomeres cultured for 110 hours after IVF, there was no significant difference between coculture and non-coculture groups(87.2% vs. 78.7%). However, the mean cell number of embryos developed from the biopsied blastomeres was significantly higher in coculture group(11.5±4.7 vs. 5.9±1.9, p<0.05). In conclusion, biopsy of mouse embryos after ZD with ATS is a safe and highly efficient method for PGD, and coculture with Vero cells showed a positive effect on the development in vitro of biopsied mouse embryos and blastomeres as a preclinical model for PGD of human embryos.
키워드 : Preimplantation genetic diagnosis(PGD), Mouse embryo, Blastomere biopsy, Zona drilling, Coculture, V
교신저자 :
전문 파일 : 
