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Comparison of Ham's F10 with CO₂ or Hepes Buffer for Storage of Equine Embryos at 5 C for 24 h¹

Colorado State University², Fort Collins 80523

Abstract

Forty equine embryos collected 7 d post-ovulation were stored at 5 C for 24 h in one of two culture media (n=20/group): 1) Ham's F10 + 10% heat-treated fetal calf serum (FCS) buffered by gassing with 5% CO₂, 5% O₂ and 90% N₂ and 2) Ham's F10 + 10% FCS with Hepes buffer (25 mM). Embryos cultured in Ham's F10 + CO₂ maintained a better quality score and had a larger average increase in diameter (+34.8 µm) than embryos stored in Hepes buffered Ham's F10 (–10.2 µm). Embryos were transferred surgically into recipient mares that ovulated –3 to +1 d in relation to the donor mare. Twenty embryos cultured in Dulbecco's phosphate buffered saline + 10% FCS and transferred <1 h after collection were used as controls. Pregnancy rates were higher (P<.05) for embryos stored in Ham's F10 + CO₂ (70%, 55%) than for embryos stored in Ham's F10 + Hepes (20%, 15%) at 14 and 35 d, respectively. At 14 d, pregnancy rates for control embryos (90%) were similar (P>.05) to pregnancy rates for embryos cultured in Ham's F10 + CO₂ (70%); however, by 35 d, pregnancy rates were higher (P<.05) for controls (80%) than for embryos stored in Ham's F10 + CO₂ (55%). It was concluded that Ham's F10 + CO₂ was superior to Ham's F10 + Hepes for short-term storage of equine embryos at 5 C, and that satisfactory pregnancy rates could be obtained from transfer of embryos stored in Ham's F10 + CO₂ at 5 C for 24 h.

¹ Supported in part by Van Camp Foundation, Long Beach, CA; Abney Foundation, Greenwood, SC; Hamilton-Thorn Research Lab, Danvers, MA and Smith Farms, Denver, CO.

² Anim. Reprod. Lab.