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This study was carried out to evaluate the effects of embryonic stage and toxicities of cryoprotectant on the rates of survival and development of the cryopreserved mouse early embryo and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology. Toxicities of two cryoprotectants, dimethyl sulfoxide (DMSO) and ethylene glycol (EG) were investigated using a murine embryo model. Female F-1 mice were stimulated with gonadotropin, induced ovulation with hCG and mated. Two cell embryos were collected and cultured after exposure to either DMSO or EG. Embryo development was evaluated up to the blastocyst stage. The total cell count of blastocysts that were treated with DMSO (68.1±24.1) at the 2-cell stage was significantly lower than that were treated with EG (81.2±27.0) or the control (99.0±18.3) (p<0.001). On comparison of two cryoprotectant treated groups, the DMSO treated group showed a decreased cell count compared with the EG treated group (p<0.05). Both DMSO (15.4±1.5) and EG (10.2±1.4) treated groups showed higher apoptosis rates of cells in the blastocyst compared with the control (6.1±0.9, p<0.0001). In addition, the DMSO treated group showed more apoptotic cells than the EG treated group (p<0.001). The potential toxicity of cryoprotectants was uncovered by prolonged exposure of murine embryos to either DMSO or EG at room temperature. When comparing two cryoprotective agents, EG appeared to be less toxic than DMSO at least in a murine embryo model.
목차
서 론 재료 및 방법 실험동물의 사양관리 시약 및 배양액 수정란 준비 배 발달율 분석 통계적 분석 결 과 동결 보호제에 따른 배아 발달율 비교 분석 동결 보호제에 따른 배반포 세포수 비교 분석 동결 보호제에 따른 세포사멸 비교 분석 고 찰 요 약 감사의 글 REFERENCES