The Effect of Coculture with Human Amniocyte on Growth and Development of Mouse 1-cell Stage Mouse Embryos

Kim, H M

Abstract

OBJECT: To examine the effect of coculture with human amniocyte on growth and development of 1-cell stage mouse embryo. Matrial and
METHODS
F1 hybrid mice was superovlated with PMSG/hCG and 1-cell stage mouse embryos were recruited. And 1-Cell stage mouse embryos were cocultured on amniocyte and Vero cell with m-KRB and Ham`s F-10. After then examined the development to 2-Cell stage mouse embryos, blastocysts and hatching blastocysts.
RESULTS
When 1-cell stage mouse embryos were cultured by using m-KRB media without coculture (control), with amniocyte, and with Vero cell, the development rates to 2-Cell stage mouse embryo were 100%, 100%, 100%, respectively. So there were no significant difference between each group. The development rates to blastocyst were 76.2%, 67.6%, 82.5%, respectively, and there were no significant difference between each group. And the rates to hatching blastocyst were 11.9%, 27.0%, 47.5%, respectively, and the coculture group with amniocyte and the control gorup (p<0.05), the control and the coculture group with Vero cell (p<0.01), and the coculture group with amniocyte and the coculture group with Vero cell (p<0.01) was signigicant defference, respectively. When 1-cell stage mouse embryos were cultured by using Ham`s F-10 media without coculture (Control), with amniocyte, and with Vero cell, the development rates to 2-Cell stage mouse embryo were 90.7%, 97.9%, 97.7% respectively. So there were no significant difference between each group. The development rates to blastocyst were 60.5%, 63.8%, 58.1%, respectively, and there were no significant difference between each group. And the rates to hatching blastocyst were 25.6%, 12.8%, 20.9%, respectively and there were no significant difference between both group. When the comparence with the development by media, (simple or complex), the developmental rates to 2-Cell embryos in control group was 100% in m-KRB media, and 90.7% in Ham`s F-10 media, and the group in m-KRB was the significantly higher difference than Ham`s F-10 (p<0.05). And the developmental rates to blastocyst was 76.2% and 60.5%, respectively, and there was no significant difference between both group. And the developmental rates to hatching blastocyst was 11.9%, and 25.6% respectively, and there was no significant difference between both group. When the comparence with the developmental rates to 2-cell embryos in coculture with human amniocyte group was 100% in m-KRB, and 97.9% in Ham`s F-10 media, and there was no significant difference between both gorup. And the developpmental rates to blastocyst was 67,6% and 63.8%, respectively, and there were no significant difference between both group. The developmental rates to hatching blastocyst was 27.0%, and 12.8%, respectively, and there was no significant difference between both group. When the comparence with the developmental rates to 2-cell embryos in coculture with Vero cell group was 100% in m-KRB, and 97.7% in Ham`s F-10 media, and there was no significant difference between both group. And the developmental rates to blastocyst was 82.5% in m-KRB, and 58.1% in Ham`s F-10 media, and the group in m-KRB was the significantly higher difference Ham`s F-10 group (p<0.05). And the developmental rates to hatching blastocyst was 47.5% in m-KRB media, and 20.9% in Ham`s F-10 media, and the group in m-KRB has the signigicantly hifher difference than Ham`s F-10 (p<0.05).
CONCLUSION
As the fact of the result, there is no coculture effect in using the complex media, and the coculture effect mainly seems to be to the late development of preimplantational embryos. Coculture effect with amniocyte is not better than with Vero cell. So therefore, reproduction related cells do not always have favor effect on in vitro growth and development. The coculture condition which induces the best effect on in vitro growth and development might be the mimics of the physiologic natural condition, just like the coculture with the tubal epithelium.