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BASIC STUDIES FOR IMPROVEMENT OF IN VITRO FERTILIZATION AND CRYOPRESERVATION

改进体外受精和冷冻保存的基础研究

基本信息

批准号：
10671518
负责人：
TSUTSUMI Osamu
金额：
$ 2.18万
依托单位：
UNIVERSITY OF TOKYO
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671518/
关键词：
embryo cryopreservation in vitro fertilization glucose vitirification GLUT implantation cryopreservatiopn vitirification dioxin estrogen receptor

项目摘要

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.

比较了两种冷冻方法（常规慢速控制速率冷冻和直接投入液氮玻璃化冷冻）对小鼠2-细胞胚胎及其囊胚发育的影响，并将新鲜和冷冻的小鼠2-细胞胚胎在体外发育为囊胚。玻璃化冷冻胚胎发育成囊胚的比例显著低于新鲜胚胎和慢速控制速率冷冻胚胎。虽然从每个冷冻保存程序的囊胚出现形态正常，无论是在囊胚中的细胞数量，也没有在体外滋养层扩散显着不同，有显着差异，在他们的功能活力。首先，玻璃化冷冻和解冻的2-细胞胚胎中葡萄糖掺入活性（[3 H]2-脱氧葡萄糖（2-DG）摄取）与新鲜或慢速控制速率冷冻和解冻的2-细胞胚胎相比显著降低。第二，从新鲜2-细胞胚胎和从慢速控制速率冷冻或玻璃化冷冻的2-细胞胚胎体外发育的囊胚的2-DG摄取分别为105 +/- 75、43.0 +/- 28.3和22.0 +/- 11.4 fmol/胚胎/h。玻璃化冷冻2-细胞胚胎体外发育的囊胚植入率（10.2%）显著低于新鲜2-细胞胚胎（30.8%）和慢速冷冻2-细胞胚胎（22.1%）。由于这些数据表明，冷冻保存可能会对胚胎的功能发育产生深远的影响，玻璃化冷冻可能会产生更有害的影响比缓慢控制速率冷冻，更多的注意，在玻璃化冷冻是常规使用在临床计划之前，其安全性。