Evaluation of Capsule Permeability in the Equine Blastocyst

Recipient pregnancy rates following transfer of frozen-thawed blastocyst stage equine embryos remain low. To date, no protocol has been developed that would allow successful cryopreservation of this stage of embryos. These experiments characterized the amount of glycerol entering equine embryos after incubation in 1.4 M and 3.4 M glycerol solutions using tritiated glycerol and a liquid scintillation counter. Blastocyst stage equine embryos (n = 27) were collected and incubated for 15 minutes in either 1.4 M (n = 14) or 3.4 M (n = 13) tritiated glycerol solutions. Disintegrations per minute were then determined, and the percent glycerol uptake was calculated for each embryo. Percent glycerol uptake for 1.4 M or 3.4 M glycerol treatment groups was not different (P = .68). However, it was higher (P = .05) in embryos with a diameter of ≤600 μm (3.6%) compared with embryos with a diameter of >600 μm (0.4%). We concluded that glycerol more readily permeates into embryos with a diameter of ≤600 μm that do not possess a fully functional capsule compared with embryos with a diameter of >600 μm with a fully formed capsule.