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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.
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.