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 analysed 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. Immunohistochemical 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.
Heald
K A
KA
Charles Hodgson Islet Laboratory, University Department of Surgery, Queen Elizabeth Hospital, Edgbaston, Birmingham, UK.
Hail
C A
CA
Hurst
R P
RP
Kane
N
N
Downing
R
R
D000818:Animals; D002241:Carbohydrates; D002469:Cell Separation / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D016650:Fluorescein-5-isothiocyanate; D007515:Islets of Langerhans / Q000166:cytology*; D037102:Lectins; D013552:Swine
D002241:Carbohydrates; D037102:Lectins; D016650:Fluorescein-5-isothiocyanate