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Updated: Jun 11, 2026
In Vitro Colony Assays for Characterizing Tri-potent Progenitor Cells Isolated from the Adult Murine Pancreas
Published on: June 10, 2016
Characterization of human pancreatic progenitor cells
Hirofumi Noguchi1, Bashoo Naziruddin, Andrew Jackson
1Baylor All Saints Medical Center, Baylor Research Institute, Fort Worth, TX 76104, USA. hirofumn@baylorhealth.edu
Abstract:
β-Cell replacement therapy via islet transplantation is an effective treatment for diabetes mellitus, but its widespread use is severely limited by the shortage of donor organs. Because pancreatic stem/progenitor cells are abundantly available in the pancreas of these patients and in donor organs, the cells could become a useful target for β-cell replacement therapy. We previously established a mouse pancreatic stem cell line without genetic manipulation. In this study, we used the techniques to identify and isolate human pancreatic stem/progenitor cells. The cells from a duct-rich population were cultured in 23 kinds of culture media, based on media for mouse pancreatic stem cells or for human embryonic stem cells. The cells in serum-free media formed "cobblestone" morphologies, similar to a mouse pancreatic stem cell line. On the other hand, the cells in serum-containing medium and the medium for human embryonic stem cells formed "fibroblast-like" morphologies. The cells divided actively until day 30, and the population doubling level (PDL) was 6-10. However, the cells stopped dividing after 30 days in any culture conditions. During the cultures, the nucleus/cytoplasm (N/C) ratio decreased, suggesting that the cells entered senescence. Exendin-4 treatment and transduction of PDX-1 and NeuroD proteins by protein transduction technology into the cells induced insulin and pancreas-related gene expression. Although the duplications of these cells were limited, this approach could provide a potential new source of insulin-producing cells for transplantation.
Insights
Researchers identified and isolated human pancreatic stem/progenitor cells for diabetes treatment. These cells, when cultured and treated, showed potential for generating insulin-producing cells, offering a new avenue for cell replacement therapy.
Area of Science:
- Stem cell biology
- Endocrinology
- Regenerative medicine
Background:
- Islet transplantation is an effective diabetes mellitus treatment but limited by donor organ scarcity.
- Pancreatic stem/progenitor cells are abundant and represent a potential source for β-cell replacement therapy.
- Previous work established a mouse pancreatic stem cell line without genetic manipulation.
Purpose of the Study:
- To identify and isolate human pancreatic stem/progenitor cells.
- To investigate the culture conditions and characteristics of these cells.
- To explore methods for inducing differentiation into insulin-producing cells.
Main Methods:
- Isolation of cells from a duct-rich human pancreatic population.
- Culture in 23 different media, including serum-free and human embryonic stem cell media.
- Analysis of cell morphology, proliferation (population doubling level), and senescence.
- Treatment with Exendin-4 and protein transduction of PDX-1 and NeuroD.
Main Results:
- Cells cultured in serum-free media exhibited cobblestone morphology, while others showed fibroblast-like morphology.
- Proliferation occurred up to day 30 with a population doubling level of 6-10, followed by senescence.
- Exendin-4 and protein transduction induced insulin and pancreas-related gene expression.
Conclusions:
- Human pancreatic stem/progenitor cells can be isolated and cultured, though their proliferative capacity is limited.
- Treatment with Exendin-4 and specific proteins can induce differentiation markers.
- This approach offers a potential new source of insulin-producing cells for diabetes therapy, despite proliferation limitations.
