Format

Send to

Choose Destination
Sci Rep. 2015 Sep 2;5:13681. doi: 10.1038/srep13681.

Rapid, high efficiency isolation of pancreatic ß-cells.

Author information

1
Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.
2
Division of Immunology, Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts.
3
Evergrande Center for Immunologic Diseases, Harvard Medical School and Brigham and Women's Hospital, Boston, Massachusetts.
4
Department of Systems Biology, Harvard Medical School, Boston, Massachusetts.

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.

PMID:
26330153
PMCID:
PMC4557033
DOI:
10.1038/srep13681
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center