Isolation and culture of single 8-cell blastomeres in-rabbits with a novel approach

Abstract

This study was undertaken to develop a new technique to produce identical offspring by aggregating a quarter or eighth embryo with a parthenogenetically activated egg in the mouse. One or two blastomeres from 8-cell embryos were aggregated with a parthenogenetic 4-cell egg from which one or two blastomeres had been removed. After micromanipulation and culture for 2 d in vitro, the morphologically normal blastocysts were transferred to the uterus of recipient females. The success rate in micromanipulation of eggs was 93 to 100%: aggregation of blastomeres occured about 60% of the time and the proportion of live young after transfer of aggregated eggs was 11 to 33% for the quarter and 2 to 24% for the eighth egg. The proportion of chimaeras as judged by coat color was 10% for the quarter and 20% for the eighth egg. However, GPI-1 analysis and progeny testing could not detect a parthenogenetic contribution in all offspring. The mean number of young obtained from one embryo was 1.7 for the quarter and 1.6 for the eighth embryo. The maximal number of young obtained from splitting one 8-cell embryo into quarters was three and into eighths was four. The mice of each set derived from a single embryo were of the same sex.Our study clearly demonstrates that the parthenogenone can assist development of the quarter and eighth mouse embryo to term. The proportion of chimaeras is low compared with that obtained when two fertilied eggs are combined.