Abstract
Progenitor cells in the adult pancreas are potential sources of endocrine beta cells for treating type 1 diabetes. Previously, we identified tri-potent progenitor cells in the adult (2–4month-old) murine pancreas that were capable of self-renewal and differentiation into duct, acinar, and endocrine cells in vitro. These progenitor cells were named pancreatic colony-forming units (PCFUs). However, because PCFUs are a minor population in the pancreas (~1%) they are difficult to study. To enrich PCFUs, strategies using cell-surface marker analyses and fluorescence-activated cell sorting were developed. We found that CD133highCD71low cells, but not other cell populations, enriched PCFUs by up to 30 fold compared to the unsorted cells. CD133highCD71low cells generated primary, secondary, and subsequent colonies when serially re-plated in Matrigel-containing cultures, suggesting self-renewal abilities. In the presence of a laminin hydrogel, CD133highCD71low cells gave rise to colonies that contained duct, acinar, and Insulin+Glucagon+ double-hormonal endocrine cells. Colonies from the laminin hydrogel culture were implanted into diabetic mice, and five weeks later duct, acinar, and Insulin+Glucagon− cells were detected in the grafts, demonstrating tri-lineage differentiation potential of CD133highCD71low cells. These CD133highCD71low cells will enable future studies of putative adult pancreas stem cells in vivo.
Highlights
The pancreas plays a critical role in regulating metabolism and is composed of three major cell lineages: acinar cells, which secrete digestive enzymes such as amylase; duct cells, which transport digestive enzymes into the gut and secrete mucin to fend off pathogens; and endocrine cells, which secrete hormones such as insulin and glucagon that are important for glucose homeostasis
We report that sorting for CD133highCD71low cells enriches pancreatic colony-forming units (PCFUs) progenitors that form Ring or 1° Endocrine/Acinar colonies in Matrigel or laminin hydrogel, respectively
The enrichment of these PCFUs is achieved using a strategy that has been successful in purifying adult hematopoietic stem cells (HSCs) [37]
Summary
The pancreas plays a critical role in regulating metabolism and is composed of three major cell lineages: acinar cells, which secrete digestive enzymes such as amylase; duct cells, which transport digestive enzymes into the gut and secrete mucin to fend off pathogens; and endocrine cells, which secrete hormones such as insulin and glucagon that are important for glucose homeostasis.Pancreas development follows a sequence of events regulated by transcription factors. Cells expressing markers for acinar (Ptf1a [14, 15], carboxypeptidase A [16], or elastase [15]) or endocrine cells (insulin [9] or glucagon [17]) have been shown to give rise to new beta cells in the adult murine pancreas. Together, these results suggest that there are multiple mechanisms by which beta cells can be replenished from different cellular sources in the adult pancreas.
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Published Version
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