Purification of beta cells from rat islets by monoclonal antibody-fluorescence flow cytometry.
Fluoresceinated monoclonal antibody plus flow cytometry was used to purify beta cells from mixed pancreatic islet endocrine cell populations. A2B5, a monoclonal antibody to a glycolipid on the surface of cells of neuroendocrine origin, was incubated with single cells dissociated from rat pancreatic islets. Antibody-bound cells were labeled with fluoresceinated goat F(ab')2 antimouse immunoglobulin and highly fluorescent cells were separated from less fluorescent cells on a Coulter EPICS IV cell sorter. Sorted cell populations were analyzed by radioimmunoassay for insulin, glucagon, and somatostatin. The highly fluorescent cell population was enriched sixfold for insulin-containing beta cells, indicating that islet beta cells are relatively enriched in A2B5 antigen and can be partially purified by this method.
D000818:Animals; D000911:Antibodies, Monoclonal; D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D006017:Glycolipids / Q000276:immunology; D007328:Insulin / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D051381:Rats; D011920:Rats, Inbred WF
D000911:Antibodies, Monoclonal; D006017:Glycolipids; D007328:Insulin