Effects of embryo density and co-culture of unfertilized oocytes on embryonic development of in-vitro fertilized mouse embryos.

We have evaluated the effects of embryo density and the co-culture of unfertilized (degenerating) oocytes on the development of in-vitro fertilized (IVF) mouse embryos. In experiment 1, groups of one, five, 10 or 20 zygotes were cultured in 20 microliter drops of modified human tubal fluid (HTF) medium for 168 h at 38.7 degrees C in 5% CO2 and 95% air. As the embryo density increased, significantly (P < 0.05) higher rates of embryos reached hatched blastocyst stage. In addition, the time required for hatching after IVF was significantly (P < 0.05) shortened by the increase in embryo density. In experiment 2, 10 IVF zygotes were cultured with or without 10 unfertilized (degenerating) oocytes in 20 microliter drops of HTF medium. The rates of IVF embryos that developed to morula, blastocyst, expanded blastocyst and hatched blastocyst stages were decreased significantly (P < 0.01) by culturing embryos with unfertilized oocytes compared with culturing embryos alone. In experiment 3, groups of one or 10 IVF zygotes or 10 IVF zygotes plus 10 unfertilized oocytes were cultured in 20 microliter drops of HTF medium and the number of cells per blastocyst was examined at 120 h after IVF. Increasing embryo density resulted in a significant (P < 0.05) increase in the number of cells per blastocyst. In contrast, the cell number of IVF embryos that developed to blastocyst decreased significantly (P < 0.05) when they were cultured with unfertilized oocytes. The results suggest that in-vitro development of IVF mouse embryos is enhanced by increasing embryo density and is impaired by co-culture with unfertilized (degenerating) oocytes.