Exposure of bovine embryos to trypsin during washing does not decrease embryonic survival

The objective of this study was to assess whether the exposure of zona pellucida-intact bovine embryos to the proteolytic enzyme, trypsin, during embryo washing has a detrimental effect on their subsequent survival and development. Embryos were collected nonsurgically from superovulated cows (n = 19) 7.5 d after insemination. Grade 1 and Grade 2 embryos were washed 12 times in modified Dulbecco's phosphate buffered saline (PBS) containing 0.4% bovine serum albumin (BSA), or in a series of five washes in BSA-PBS (without Ca++ and Mg++), two in 0.25% trypsin in Hank's solution (without Ca++ and Mg++) and five in PBS-BSA medium. Within 30 min after washing, embryos were either transferred nonsurgically into recipient cows, 7 to 8 d post estrus, or cryopreserved and transferred later. Frozen-thawed embryos from five of the donors were cultured for 72 h in vitro and their development was evaluated. Pregnancy rates did not differ (P>0.1) between recipient cows receiving control-washed and trypsin-washed embryos transferred fresh (51.0 vs 56.3%). However, pregnancy rates were higher (P<0.05) for frozen-thawed embryos treated with trypsin before cryopreservation than for frozen-thawed, control-washed embryos (68.2 vs 38.5%). Survival and development of embryos in vitro after cryopreservation did not differ between embryos subjected to the control- and trypsin-wash procedures. These results suggest that exposure of bovine embryos to trypsin for 2 to 3 min during washing did not have a detrimental effect on embryonic development, but may have enhanced cryopreservation of the embryos.