Basic study in cryopreservation and chromosomal analysis of single blastomeres.

单卵裂球冷冻保存和染色体分析的基础研究。

基本信息

批准号：
02670728
负责人：
SENGOKU Kazuo
金额：
$ 1.41万
依托单位：
ASAHIKAWA MEDICAL COLLEGE
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1992
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02670728/
关键词：
embryobiopsy cryopreservation chromosomal analysis blastomeres

项目摘要

Biopsy of the preimplantation embryo has wide application as a tool to investigate the developmental potential of the preimplantation embryo and also offered a possibility for prenatal diagnosis. The purpose of this study were first, to examine the developmental potential of single blastomeres of four-and eight-cell mouse embryos, and second to investigate the possibility of successful cryopreservation and karyotyping of single blastomeres.(1) Removal of a single blastomere from 4- and 8-cell mouse embryos by extrusion technique did not affect development to the blastocyst stage in vitro (80% of control,75% and 78.3% of biopsied embryo at 4- and 8-cell stage, respectively). However,after transfer to pseudopregnant recipient mice, the implantation rates of biopsied embryos were significantly lower than intact-controls (24.3% at 4-cell, 34.8% at 8-cell and 50% at control, respectively).(2) Most single blastomeres had undergone several cell divisions in vitro. After a short culture of single blastomeres with subsequent exposure to colcemid for 16 hours, 27.3% of 4-cell and 52.2% of 8-cell biopsies were karyotyped. However, 20 to 30% of biopsied blstomeres were lost through cell death and technical problems.(3) Biopsied single blastomeres could not be successfully cryopreserved when the freezing medium contained 1.5M DMSO+0.25M sucrose and 5-step dilution method was used. The techniques developed in this study may provide the useful tool for the study of prenatal diagnosis.

植入前胚胎的活检具有广泛的应用，是研究植入前胚胎的发育潜力的一种工具，还提供了产前诊断的可能性。 The purpose of this study were first, to examine the developmental potential of single blastomeres of four-and eight-cell mouse embryos, and second to investigate the possibility of successful cryopreservation and karyotyping of single blastomeres.(1) Removal of a single blastomere from 4- and 8-cell mouse embryos by extrusion technique did not affect development to the blastocyst stage in vitro (80% of control,75% and 78.3% of biopsied分别在4个和8细胞阶段的胚胎）。但是，在转移到伪久的受体小鼠后，活检胚胎的植入率显着低于完整控制的（4细胞时24.3％，在8细胞时为34.8％，对照组分别为50％）。（2）大多数单个单胚层的植入率都在维持了几个单元格。在短期培养单个胚泡并随后暴露于Colcemid持续16小时后，对4细胞的27.3％和52.2％的8细胞活检进行了核分型。然而，通过细胞死亡和技术问题，有20％至30％的活检蓝藻丢失了。（3）当冻结培养基含有15m dmso+0.25万蔗糖和5步稀释方法时，活检的单个胚泡无法成功保存。这项研究中开发的技术可能为研究产前诊断提供了有用的工具。