SEPARATION OF BETA-CELLS FROM DISPERSED PORCINE PANCREAS BY SELECTIVE LECTIN BINDING

Variation in cell-surface sugar residues which exist between different pancreatic cells has been exploited in an attempt to isolate beta-cells from dispersed porcine pancreas utilizing selective lectin binding. The binding characteristics of a range of lectins were compared to determine their ability to differentiate between endocrine and non-endocrine cells in the porcine pancreas. Histological analysis showed that peroxidase labelled Arachis hypogaea bound selectively to islet cells in Carnoy-fixed sections of pancreas. In five experiments, porcine pancreas was dispersed into single cells by collagenase digestion, incubated with fluorescein isothiocyanate-labelled Arachis hypogaea and analyses using a Fluorescence Activated Cell Sorter. Fluorescein isothiocyanate-labelled Arachis hypogaea bound to a population of cells comprising 6% +/- 4.2% (mean +/- s.d.) of the total. Cells from representative samples were sorted into populations, based on fluorescence. Immuno-histochemical analysis of the fluorescent populations showed that 93% +/- 2% of these cells contained insulin: none of the cells stained positive for glucagon or somatostatin. These preliminary studies show that it is possible to separate porcine beta-cells from a dispersed cell preparation using a fluorescent labelled lectin.