Release of chylomicrons by isolated cells of rat intestinal mucosa

Abstract

Fat-laden mucosal cells were isolated by flotation from fed male rats after digesting scrapings of washed jejunum with collagenase in bicarbonate buffer. About 50–60 million cells were obtained per preparation, which were 95–100% viable as assessed by Trypan Blue. The isolated cells were capable of effective incorporation of labeled fatty acids and glucose into triglycerides and phospholipids, and of labeled leucine and glucosamine into the protein envelope of the released chylomicrons. The secretion of the labeled protein paralleled the release of the labeled fat, both of which were linear with the concentration of the albumin in the incubation mixture. About 80% of the total fat of the cell was released as chylomicrons within 30 min when incubated in the presence of albumin-bicarbonate buffer. Injection of puromycin 24 hr prior to harvesting of cells led to a complete inhibition of chylomicron release. Addition of puromycin to the incubation medium gave 50–80% inhibition of release. No inhibition of release of chylomicrons resulted from a treatment with ethionine. The released chylomicrons were separated from the cells by Millipore filtration.