Rapid, high efficiency isolation of pancreatic ß-cells.
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.
Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.
Susan M
SM
Clardy
Division of Immunology, Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts.
James F
JF
Mohan
Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.
Claudio
C
Vinegoni
Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.
Edmund J
EJ
Keliher
Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.
Yoshiko
Y
Iwamoto
Division of Immunology, Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts.|Evergrande Center for Immunologic Diseases, Harvard Medical School and Brigham and Women's Hospital, Boston, Massachusetts.
Christophe
C
Benoist
Division of Immunology, Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts.|Evergrande Center for Immunologic Diseases, Harvard Medical School and Brigham and Women's Hospital, Boston, Massachusetts.
Diane
D
Mathis
Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.|Department of Systems Biology, Harvard Medical School, Boston, Massachusetts.
Ralph
R
Weissleder
D000818:Animals; D001896:Boron Compounds / Q000378:metabolism; D002469:Cell Separation / Q000379:methods*; D003921:Diabetes Mellitus, Experimental / Q000235:genetics / Q000473:pathology; D000077270:Exenatide; D005260:Female; D020869:Gene Expression Profiling; D005934:Glucagon / Q000378:metabolism; D007328:Insulin / Q000378:metabolism; D050417:Insulin-Secreting Cells / Q000166:cytology*; D008810:Mice, Inbred C57BL; D016688:Mice, Inbred NOD; D010455:Peptides / Q000378:metabolism; D013004:Somatostatin / Q000378:metabolism; D014688:Venoms / Q000378:metabolism
D001896:Boron Compounds; D007328:Insulin; D010455:Peptides; D014688:Venoms; D013004:Somatostatin; D005934:Glucagon; D000077270:Exenatide