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Beneficial effects of culturing rabbit zygotes to blastocysts in 5% oxygen and 10% carbon dioxide

Zygotes were collected from superovulated Dutch-belted rabbits 19 h after injection of LH and insemination. Oocytes that appeared to be unfertilized were discarded. The zygotes were distributed equally within each donor female across all culture treatments. Culture dishes contained 500 µl of macromolecule-free RD medium consisting of equal parts of RPMI 1640 and low glucose Dulbecco's modified Eagle's medium. Embryos were cultured at 39°C in several gas combinations of N₂ plus the following: (1) 1% O₂:10% CO₂; (2) 5% O₂:10% CO₂; and (3) 20% O₂:10% CO₂. The control (4) was 95% air:5% CO₂. The experiment was replicated with embryos from 11 donors providing 295 usable zygotes. After 84 h of culture, the percentages of blastocysts formed in treatments 1 to 4, respectively, were 13, 86, 82 and 59 (P < 0.01). The corresponding mean cell counts, including all cleaved embryos cultured (but not degenerate ones), were 55, 183, 118 and 68 (P < 0.01). These results indicate that 10% CO₂ combined with 5% O₂ is a more effective gas phase for culturing rabbit zygotes in a synthetic medium than is the commonly used 5% CO₂, and that 5% O₂ is superior to either 1% or 20% O₂.

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