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Commercial samples of bovine serum albumin (BSA) in a complex medium caused growth of 1-cell rabbit embryos to completely hatched blastocysts. Heat treatment of the BSA at 65 or 80 degrees C significantly decreased blastocyst formation and expansion and destroyed the ability to cause blastocyst hatching. Addition of trypsin at levels down to 20 ng/ml caused the formation of hatched blastocysts which degenerated rapidly. The effects of 5 protease inhibitors (ovomucoid trypsin inhibitor, alpha-1-antitrypsin, TAME, TLCK and soybean) were tested. Ovomucoid trypsin inhibitor, TAME and TLCK significantly inhibited blastocyst hatching but only at the highest concentration used. These inhibitors also reduced blastocyst formation and expansion, indicating that their effect was not specifically on blastocyst hatching in vitro. It is concluded that hatching of rabbit blastocysts is probably not dependent on protease action.

Evidence that protease action is not specifically involved in the hatching of rabbit blastocysts caused by commercial bovine serum albumin in culture