Cells with surface expression of CD133^(high)CD71^(low) are enriched for tripotent colony-forming progenitor cells in the adult murine pancreas
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 (2–4 month-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 CD133^(high)CD71^(low) cells, but not other cell populations, enriched PCFUs by up to 30 fold compared to the unsorted cells. CD133^(high)CD71^(low) 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, CD133^(high)CD71^(low) 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 CD133^(high)CD71^(low) cells. These CD133^(high)CD71^(low) cells will enable future studies of putative adult pancreas stem cells in vivo.
Additional Information
© 2015 Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Received 11 July 2014. Received in revised form 7 November 2015. Accepted 25 November 2015. Available online 30 November 2015. This work is supported in part by the National Institutes of Health (NIH) grants R01DK081587 and R01DK099734 to H.T.K. and U01DK089533 to A.D.R., National Science Foundation grant NSF-DMR-1206121 and California Institute for Regenerative Medicine grant RB5-07398 to D.A.T., and Office of Naval Research ONR-N00014-02-1 0958 and NSF-DBI-9970143 to the Electron Microscopy Core facility at City of Hope. Support from the Joseph J. Jacobs Institute for Molecular Engineering for Medicine at Caltech is also gratefully acknowledged. L.J. is supported by the National High Technology Research and Development Program of China (863 Program, No. 2015AA020314), Excellent Youth Foundation of Jiangsu Scientific Committee (BK20140029), the Fundamental Research Funds for the Central Universities (Z114037), Priority Academic Program Development of Jiangsu Higher Education Institutions, and the National Natural Science Foundation of China (No. 81570696). Research reported in this publication included work performed in the Analytical Cytometry Core and Light Microscopy Digital Imaging Core supported by the National Cancer Institute of the National Institutes of Health under award number P30CA33572. The sponsor did not participate in the study design, collection, analysis, or interpretation of data. Author Responsibilities: Liang Jin, Dan Gao: study concept and design, acquisition of data, analysis and interpretation of data, drafting of the manuscript. Tao Feng, Angela Luo, Jeffery Rawson, Janine Quijano, Jing Chai, Lena Wedeken, Nadiah Ghazalli, Jasper Hsu, Jeanne LeBon, Stephanie Walker, Jacob Tremblay: acquisition of data, analysis and interpretation of data. Alborz Mahdavi, Hung-Ping Shih: technical or material support. David A. Tirrell, Arthur D. Riggs: technical or material support, critical revision of the manuscript for important intellectual content, obtained funding. Hsun Teresa Ku: study concept and design, analysis and interpretation of data, drafting of the manuscript, critical revision of the manuscript for important intellectual content, obtained funding. The authors have nothing to disclose.Attached Files
Published - 328-Jin-2015.pdf
Accepted Version - nihms745580.pdf
Supplemental Material - mmc1.docx
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328-Jin-2015.pdf
Additional details
Identifiers
- PMCID
- PMC4762724
- Eprint ID
- 63344
- Resolver ID
- CaltechAUTHORS:20160104-143534788
Funding
- NIH
- R01DK081587
- NIH
- R01DK099734
- NIH
- U01DK089533
- NSF
- DMR-1206121
- California Institute for Regenerative Medicine (CIRM)
- RB5-07398
- Office of Naval Research (ONR)
- N00014-02-1 0958
- NSF
- DBI-9970143
- Joseph J. Jacobs Institute for Molecular Engineering for Medicine
- National High Technology Research and Development Program of China
- 2015AA020314
- Excellent Youth Foundation of Jiangsu Scientific Committee
- BK20140029
- Fundamental Research Funds for the Central Universities
- Z114037
- Priority Academic Program Development of Jiangsu Higher Education Institutions
- National Natural Science Foundation of China
- 81570696
- National Cancer Institute
- P30CA33572
Dates
- Created
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2016-01-04Created from EPrint's datestamp field
- Updated
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2022-05-16Created from EPrint's last_modified field
Caltech Custom Metadata
- Caltech groups
- Jacobs Institute for Molecular Engineering for Medicine