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Diabetes. 2015 Sep;64(9):3172-81. doi: 10.2337/db15-0039. Epub 2015 Apr 30.

Novel Observations From Next-Generation RNA Sequencing of Highly Purified Human Adult and Fetal Islet Cell Subsets.

Author information

1
Department of Medicine, Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, MA.
2
Program in Molecular Medicine, Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, MA.
3
Program in Molecular Medicine, Program in Bioinformatics, University of Massachusetts Medical School, Worcester, MA.
4
Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA.
5
Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA.
6
Program in Molecular Medicine, University of Massachusetts Medical School, and Howard Hughes Medical Institute, Worcester, MA.
7
Department of Medicine, Diabetes Center of Excellence, University of Massachusetts Medical School, Worcester, MA david.harlan@umassmemorial.org.

Abstract

Understanding distinct gene expression patterns of normal adult and developing fetal human pancreatic α- and β-cells is crucial for developing stem cell therapies, islet regeneration strategies, and therapies designed to increase β-cell function in patients with diabetes (type 1 or 2). Toward that end, we have developed methods to highly purify α-, β-, and δ-cells from human fetal and adult pancreata by intracellular staining for the cell-specific hormone content, sorting the subpopulations by flow cytometry, and, using next-generation RNA sequencing, we report the detailed transcriptomes of fetal and adult α- and β-cells. We observed that human islet composition was not influenced by age, sex, or BMI, and transcripts for inflammatory gene products were noted in fetal β-cells. In addition, within highly purified adult glucagon-expressing α-cells, we observed surprisingly high insulin mRNA expression, but not insulin protein expression. This transcriptome analysis from highly purified islet α- and β-cell subsets from fetal and adult pancreata offers clear implications for strategies that seek to increase insulin expression in type 1 and type 2 diabetes.

PMID:
25931473
PMCID:
PMC4542439
DOI:
10.2337/db15-0039
[Indexed for MEDLINE]
Free PMC Article

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