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FERTILIZATION OF RABBIT EGGS IN VITRO WITHOUT SUPPLEMENTAL CO₂ IN THE ATMOSPHERE

To achieve fertilization of mammalian eggs in vitro, the gametes are often mixed and incubated in a CO₂-enriched atmosphere, e.g. rabbit (Brackett & Williams, 1968), mouse (Cross & Brinster, 1970), human (Edwards, Steptoe & Purdy, 1970). Since the various media are usually buffered with relatively high levels of bicarbonate, the CO2 is needed to maintain a pH which will allow sperm penetration. Bicarbonate has a stimulatory effect on sperm metabolism (Murdoch & White, 1971) and may facilitate capacitation of hamster spermatozoa in vitro (Bavister, 1969). Furthermore, replacement of bicarbonate with tris or phosphate in a defined medium for ovum culture resulted in cessation of development (Brinster, 1969).

Using the technique of Brackett & Williams (1968) in which eggs and spermatozoa are cultured for 4 hr in an atmosphere of 5% CO₂:95% air, it has been possible to achieve a high rate of fertilization of rabbit eggs