Primary cultures of feline acinar cells: dissociation, culturing, and viral infection

Abstract

A technique for dissociating and plating of feline parotid acinar cells by enzymatic dispersion with collagenase and trypsin is presented. Viability of the cultured cells was determined by: 1) estimating cellular growth with visual cell counts and by [3H]thymidine incorporation; 2) observing the lack of uptake of vital dyes by the cultured cells: 3) making light, phase contrast, and electron microscopic morphological examinations; and 4) determining the lack of amylase secretion without stimulation, and the evidence of amylase secretion in response to isoproterenol. The cultured feline parotid acinar cells were infected with feline rhinotracheitis virus, a member of the herpes group. Evidence for viral infection was morphological and by indirect fluorescent antibody studies. The viral infected cells also showed no amylase release without stimulation and demonstrated an initial response to a beta-adrenergic agonist by secretion of amylase, but repeated stimulation of the virally infected cells did not produce amylase secretion.