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.

比较两种冷冻保存程序（使用可编程冷冻机的传统慢速控速冷冻和直接投入液氮的玻璃化冷冻）对 2 细胞胚胎及其随后发育为囊胚的影响，新鲜或冷冻保存的 2 细胞小鼠胚胎发育为囊胚。体外囊胚。玻璃化冷冻胚胎发育成囊胚的百分比显着低于新鲜胚胎和慢控速冷冻胚胎。尽管每次冷冻保存过程中的囊胚形态均正常，且囊胚中的细胞数量和体外滋养层扩散均无显着差异，但其功能活力却存在显着差异。首先，与新鲜或缓慢控制速率冷冻和解冻的2细胞胚胎相比，玻璃化和解冻的2细胞胚胎中的[(3)H]2-脱氧葡萄糖(2-DG)摄取方面的葡萄糖掺入活性显着降低。其次，新鲜 2 细胞胚胎和缓慢控制速率冷冻或玻璃化 2 细胞胚胎体外发育的囊胚对 2-DG 的摄取量为 105 +/- 75、43.0 +/- 28.3 和 22.O +/- 11.4分别为 fmol/胚胎/h。第三，玻璃化2细胞胚胎体外发育的囊胚着床率（10.2%）显着低于新鲜2细胞胚胎（30.8%）或慢控速冷冻2细胞胚胎（22.1%）。由于这些数据表明冷冻保存可能对胚胎的功能发育产生不可挽回的影响，并且玻璃化冷冻可能比缓慢的控速冷冻产生更有害的影响，因此在临床方案中常规使用玻璃化冷冻之前，应更多地关注其安全性。