Rapid, high efficiency isolation of pancreatic ß-cells

快速、高效分离胰岛β细胞

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作者:Susan M Clardy ,James F Mohan ,Claudio Vinegoni ,Edmund J Keliher ,Yoshiko Iwamoto ,Christophe Benoist ,Diane Mathis ,Ralph Weissleder

Abstract

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.

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