Enzyme separation techniques for the study of growth of cells from layers of bovine dental pulp

Summary

Effects of the enzymes trypsin, papain, bromelains and ficin in bovine dental pulp tissue were studied. Minced or whole pulps were subjected to each enzyme at 17°, 20° and 37°C for set time intervals, after which aliquots of supernatant fluid were removed for cell counts and viability tests. Pooled samples were subsequently cultured as monolayers in Eagle’s MEM plus 10% calf serum. The dissociation characteristics were quite distinct for each enzyme, although quite similar between minced and whole pulp. A parallel histological study was made of the residual pulp tissue. Ficin was found to be the most suitable enzyme for future studies on the growth of isolated pulp cells from various layers of the bovine pulp, due to its even rate of cell removal, and the good initial viability and subsequent growth of the separated cells in monolayer culture. Further studies on ficin may show that it is more suitable for enzymatic separation of tissues generally than the more commonly used trypsin, a major advantage being its use in media containing Ca²⁺ and Mg²⁺.