THE EFFECT OF DIFFERENT PROTEOLYTIC ENZYMES ON THE DISSOLUTION OF THE ZONA PELLUCIDA OF IN VITRO PRODUCED BUFFALO (BUBALUS BUBALIS) EMBRYOS

Removal of the zona pellucida is an important step in the isolation of inner cells from embryos. The zona pellucida could be removed either by mechanical means or by exposing embryos to proteolytic enzymes as the zona is made up of glycoproteins. Even though there were various methods to isolate inner cells from embryos, enzyme digestive method was the more convenient and better method. The present study was undertaken to compare the effect of different proteolytic enzymes (0.5 percent pronase, 0.5 percent trypsin and 0.5 percent papain) on zonalysis and quality of subsequently isolated inner cells/ stem cells from in vitro-produced preimplantation buffalo embryos. Out of those enzymes, 0.5 percent pronase, was found to be effective and superior. With 0.5 percent pronase, there was no damage to inner cells, whereas 0.5 percent trypsin and 0.5 percent papain resulted in observable morphological degradation of inner cells.