Sasabe Y
1st Department of Obstetrics and Gynecology, Toho University, School of Medicine, Tokyo.
Nihon Sanka Fujinka Gakkai Zasshi. 1993 Jul;45(7):650-6.
In preimplantation genetic diagnosis, it is vital that the biopsy method does not affect embryonic development and yet provides a suitable specimen for genetic diagnosis. This study investigated whether the expulsion method, a modification of the displacement and extrusion methods, could satisfy the above mentioned conditions. Mouse embryos fertilized in vitro were biopsied at the 4, 8 and 16-cell stages, and were subsequently observed for in vitro and in vivo development. The rates of blastocyst formation, implantation, and fetal growth in the 8- and 16-cell biopsy groups were not significantly different from those in the control groups, but all 3 parameters were significantly reduced in the 4-cell group. Fetal and placental development was similar to the control group when embryos were biopsied at all three stages. Single blastomeres obtained by expulsion biopsy were subjected to DNA amplification by dual PCR, and Sry and myogenin sequences were amplified. In conclusion, expulsion biopsy did not affect the development of 8- and 16-cell embryos, and the specimens obtained were adequate for DNA amplification.
在植入前基因诊断中,活检方法不影响胚胎发育且能提供适合基因诊断的样本至关重要。本研究调查了改良的移位和挤压法——排出法,是否能满足上述条件。对体外受精的小鼠胚胎在4细胞、8细胞和16细胞阶段进行活检,随后观察其体外和体内发育情况。8细胞和16细胞活检组的囊胚形成率、着床率和胎儿生长率与对照组无显著差异,但4细胞组的这三个参数均显著降低。当在所有三个阶段对胚胎进行活检时,胎儿和胎盘发育与对照组相似。通过排出活检获得的单个卵裂球经双重PCR进行DNA扩增,扩增出了Sry和肌细胞生成素序列。总之,排出活检不影响8细胞和16细胞胚胎的发育,所获得的样本足以进行DNA扩增。