Redox-mediated enrichment of self-renewing adult human pancreatic cells that possess endocrine differentiation potential

Katrina D. Linning, Mei Hui Tai, Burra V. Madhukar, C. C. Chang, Donald N. Reed, Sarah Ferber, James E. Trosko, L. Karl Olson

Research output: Contribution to journalArticlepeer-review

Abstract

Objectives: The limited availability of transplantable human islets has stimulated the development of methods needed to isolate adult pancreatic stem/progenitor cells capable of self-renewal and endocrine differentiation. The objective of this study was to determine whether modulation of intracellular redox state with N-acetyl-l-cysteine (NAC) would allow for the propagation of pancreatic stem/progenitor cells from adult human pancreatic tissue. Methods: Cells were propagated from human pancreatic tissue using a serum-free, low-calcium medium supplemented with NAC and tested for their ability to differentiate when cultured under different growth conditions. Results: Human pancreatic cell (HPC) cultures coexpressed α-amylase, albumin, vimentin, and nestin. The HPC cultures, however, did not express other genes associated with differentiated pancreatic exocrine, duct, or endocrine cells. A number of transcription factors involved in endocrine cell development including β 2, Islet-1, Nkx6.1, Pax4, and Pax6 were expressed at variable levels in HPC cultures. In contrast, pancreatic duodenal homeobox factor 1 (Pdx-1) expression was extremely low and at times undetectable. Overexpression of Pdx-1 in HPC cultures stimulated somatostatin, glucagon, and carbonic anhydrase expression but had no effect on insulin gene expression. HPC cultures could form 3-dimensional islet-like cell aggregates, and this was associated with expression of somatostatin and glucagon but not insulin. Cultivation of HPCs in a differentiation medium supplemented with nicotinamide, exendin-4, and/or LY294002, an inhibitor of phosphatidylinositol-3 kinase, stimulated expression of insulin mRNA and protein. Conclusion: These data support the use of intracellular redox modulation for the enrichment of pancreatic stem/progenitor cells capable of self-renewal and endocrine differentiation.

Original languageEnglish
Pages (from-to)e64-e76
JournalPancreas
Volume29
Issue number3
DOIs
StatePublished - Oct 2004
Externally publishedYes

Keywords

  • Albumin
  • Amylase
  • Endocrine hormones
  • Insulin
  • Pancreatic stem cells

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