Production of cloning animals by fresh and frozen-thawed nuclear transfer embryos II

Hwang, Woo-suk; Jo, Choong-ho; Lee, Chang-woo; Lee, Byeong-chun

Korean Journal of Veterinary Research 1993;33(3):547-554.

Woo-suk Hwang, Choong-ho Jo, Chang-woo Lee, Byeong-chun Lee

College of Veterinary Medicine, Seoul National University

핵이식 수정란의 동결, 융해 및 이식에 의한 클론동물의 생산 II

황우석, 조충호, 이창우, 이병천

서울대학교 수의과대학

Abstract

This study was carried out to investigate the best condition for in vitro and in vivo culture after freezing and thawing of nuclear transplant 2 cell embryos. When nuclear transplant embryos were submitted to electrofusion, the significantly higher fusion rates of 2 cell donor nuclei were achieved at the electric field strength of DC 1.5 kV/cm for 100 and $150{mu}sec$, DC 2.0 kV/cm for 100 and $150{mu}sec$ than DC 1.0 kV/cm for 100 and $150{mu}sec$(p<0.01). The significantly higher fusion rates of 4 cell donor nuclei were achievecl at DC 2.0 kV/cm for 100 and $150{mu}sec$ than DC 1.0 kV/cm for 100 and $150{mu}sec$(p<0.01). The fusion rates in 8 cell donor nuclei were 94.2~99.3%. The developmental potency to blastocyst in 2 cell donor nuclei was significantly higher in DC 2.0 kV/cm for $150{mu}sec$ treated group(p<0.01). The significantly higher developmental potency to blastocyst in 4 cell donor nuclei were achieved at the electric field strength of DC 2.0 kV/cm for $150{mu}sec$ than DC 1.5 kV/cm for 100 and $150{mu}sec$, DC 2.0 kV/cm for $100{mu}sec$ treated group(p<0.01). The develop mental potency to blastocyst in 8 cell donor nuclei was significantly higher in DC 2.0 kV/cm for $100{mu}sec$ treated group(p<0.01). The developmental potency to blastocyst after nuclear transplantation was significantly higher in 2 cell donor nuclei than in 8 cell donor nuclei(p<0.01). When the recovered embryos in normal morphology were cultured in vitro, there were no significant differences in the developmental potency to blastocyst between the freezing methods and the concentrations of cryoprotectant(p<0.01). The production rates of offspring after transfer of nuclear transplant embryos to recipient mouse were no significant difference in 2, 4 and 8 cell donor nuclei.

Key Words: Mouse, nuclear transplantation, electrofusion, cryopreservation