A self-contained culture platform using carbon dioxide produced from a chemical reaction supports mouse blastocyst development in vitro.

Elevated CO(2) is required for in vitro embryo culture to maintain proper media pH and to supply embryo metabolic pathways. As an alternative to current approaches using gas cylinders, we examined use of a chemical reaction to supply CO(2). A closed culture system was constructed and chemicals added to generate CO(2), which was then supplied to developing embryos. This system was shown to provide a stable pH (7.2-7.4) over 4 days of use. One-cell mouse embryos were cultured in the device and no difference in blastocyst formation or cell number was apparent between embryos grown in a closed system with CO(2) supplied by a chemical reaction or positive controls grown in a an open system in a CO(2) incubator. This approach provides a highly purified, inexpensive, and easily obtainable gas source and offers potential for development of new, self-contained culture platforms.