N. Fukunaga

The use of time-lapse observations to confirm pronuclei formation and fertilization

THE USE OF TIME-LAPSE OBSERVATIONS TO CONFIRM PRONUCLEI FORMATION AND FERTILIZATION. K. Nakayama, N. Fukunaga, H. Kitasaka, T. Yoshimura, F. Tamura, M. Katou, N. Aoyagi, Y. Kida, Y. Hashiba, Y. Asada. IVF Laboratory, Asada Ladies Nagoya Clinic, Nagoya, Aichi, Japan; IVF Laboratory, Asada Ladies Kachigawa Clinic, Nagoya, Aichi, Japan; Asada Institute for Reproductive Medicine, Asada Ladies Clinic, Nagoya, Aichi, Japan.

Establishing a continuous blastocyst culture system without direct observation and exchange of culture medium by employing a time-lapse incubation system

Oct4, Plac8, Cox2, and Glut1 was determined. Data were analyzed using ANOVA with Duncan’s post-hoc test and significance was defined as a P-value less than 0.05. RESULTS: Development to fully hatched blastocyst stage was significantly increased in biopsied embryos compared to LAH embryos, which was significantly increased compared to the unmanipulated controls at 24, 28 and 40 hrs post manipulation. While the percentage of fully hatched blastocysts in the LAH-cryo and biopsy-cryo groups seemed to lag in development compared to their fresh counterparts, by 40 hrs there was no significant difference. Transcript abundance for 18s rRNA, Ppia, and Oct4 were significantly higher in biopsied embryos compared to all other treatment groups. Glut1 was significantly higher in the biopsied groups compared to all other treatments except for the biopsy-cryo, which was not significantly different. Plac8 was not significantly different in any of the treatment groups. CONCLUSION: The short-term and long-term effects of embryo manipulation are still unknown. This study suggests that survivability of biopsied embryos may be improved compared to unmanipulated embryos, contrary to the belief that ‘the least amount of manipulation is best.’ Biopsied embryos, regardless of whether the embryos were cryopreserved post-manipulation, had better developmental capabilities compared to control and LAH embryos. The increase in expression of essential developmental genes further suggests that biopsied embryos are able to compensate for the insult that occurred during manipulation, and may actually have advanced faster. Supported by: Baylor Scott & White Healthcare.