Rapid, high efficiency isolation of pancreatic ß-cells

• Published in: Scientific reports Vol. 5; no. 1; p. 13681
• Main Authors: Clardy, Susan M., Mohan, James F., Vinegoni, Claudio, Keliher, Edmund J., Iwamoto, Yoshiko, Benoist, Christophe, Mathis, Diane, Weissleder, Ralph
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 02.09.2015; Nature Publishing Group
• Subjects: 631/61; 692/163; Animals; Boron Compounds - metabolism; Cell Separation - methods; Diabetes Mellitus, Experimental - genetics; Diabetes Mellitus, Experimental - pathology; Female; Gene Expression Profiling; Glucagon - metabolism; Humanities and Social Sciences; Insulin - metabolism; Insulin-Secreting Cells - cytology; Mice, Inbred C57BL; Mice, Inbred NOD; multidisciplinary; Peptides - metabolism; Science; Somatostatin - metabolism; Venoms - metabolism
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep13681

The ability to isolate pure pancreatic ß-cells would greatly aid multiple areas of diabetes research. We developed a fluorescent exendin-4-like neopeptide conjugate for the rapid purification and isolation of functional mouse pancreatic β-cells. By targeting the glucagon-like peptide-1 receptor with the fluorescent conjugate, β-cells could be quickly isolated by flow cytometry and were >99% insulin positive. These studies were confirmed by immunostaining, microscopy and gene expression profiling on isolated cells. Gene expression profiling studies of cytofluorometrically sorted β-cells from 4 and 12 week old NOD mice provided new insights into the genetic programs at play of different stages of type-1 diabetes development. The described isolation method should have broad applicability to the β-cell field.