A comparison of rat peritoneal mast cells purified using Percoll and Path-O-Cyte 4

Abstract

Rat peritoneal mast cells (RPMC) purified through Percoll and a commercially available 35% bovine serum albumin (BSA) solution (Path-O-Cyte 4) are compared over a range of criteria with mixed peritoneal cell suspensions. Both the purity and spontaneous histamine release were slightly better in RPMC isolated through Percoll and histamine recovery was considerably greater using this method as opposed to Path-O-Cyte 4. The responsitivity of cells isolated by either method to compound 48/80 and ionomycin was comparable with that seen in non-purified RPMC. However, the histamine release induced by anti-rat IgE and concanavalin A was reduced in the purified cells especially in those isolated through BSA. Histamine release induced by IgE-directed ligands was potently inhibited by theophylline, isobutyl methyl xanthine, disodium cromoglycate and nedocromil sodium in both non-purified and Percoll-purified RPMC. In total, this study has shown that highly purified and viable RPMC can be obtained through Percoll purification. This method was seen to be generally superior to that of Path-O-Cyte 4.