TY - JOUR
T1 - Characterization of the Human Pancreas Side Population as a Potential Reservoir of Adult Stem Cells
AU - Augstein, Petra
AU - Loudovaris, Thomas
AU - Bandala-Sanchez, Esther
AU - Heinke, Peter
AU - Naselli, Gaetano
AU - Lee, Lily
AU - Hawthorne, Wayne J.
AU - Góñez, L. Jorge
AU - Neale, Alana M.
AU - Vaillant, François
AU - Thomas, Helen E.
AU - Kay, Thomas W.
AU - Banakh, Ilia
AU - Harrison, Leonard C.
N1 - Funding Information:
Abbreviations: CA 19-9 – carbohydrate antigen 19-9, CFP – colony-forming potential, DMEM – Dulbecco's modified Eagle medium, FCS – fetal calf serum, FSC – forward scatter, GAD Ab – antibodies to the islet autoantigen glutamic acid decarboxylase (molecular weight, 65,000 dalton isoform), From the *Walter and Eliza Hall Institute for Medical Research; †Department of Medical Biology, University of Melbourne, Parkville, Australia; ‡Institute of Diabetes “Gerhardt Katsch,” Karlsburg; §Department of Medicine A, University Medicine Greifswald, Greifswald, Germany; ||St Vincent's Institute for Medical Research, Fitzroy; and ¶Department of Surgery, University of Sydney, Westmead Hospital, Westmead, Australia. Received for publication March 29, 2017; accepted September 13, 2017. Address correspondence to: Leonard C. Harrison, Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, 3052, Victoria, Australia (e‐mail: [email protected]). This study was supported by a Special Program Grant (465199) from the Australian National Health and Medical Research Council and Juvenile Diabetes Research Foundation International. It was also made possible through Victorian State Government Operational Infrastructure Support and Australian National Health and Medical Research Council Research Institute Infrastructure Support Scheme. L.C.H. was the recipient of a National Health and Medical Research Council Senior Principal Research Fellowship (1080887). P.A. received an Albert Renold Travel Fellowship from the European Foundation for the Study of Diabetes. The authors declare no conflict of interest. Supplemental digital contents are available for this article. Direct URL citations appear in the printed text and are provided in the HTML and PDF versions of this article on the journal’s Web site (www.pancreasjournal.com). Copyright © 2017 Wolters Kluwer Health, Inc. All rights reserved. DOI: 10.1097/MPA.0000000000000950
Publisher Copyright:
© 2017 Wolters Kluwer Health, Inc. All rights reserved.
PY - 2018/1/1
Y1 - 2018/1/1
N2 - Objectives The side population (SP) contains cells with stem cell/progenitor properties. Previously, we observed that the mouse pancreas SP expanded after pancreatic injury. We aimed to characterize the SP in human pancreas as a potential source of stem cells. Methods Human organ donor pancreata were fractionated into islets and exocrine tissue, enriched by tissue culture and dispersed into single cells. Cells were phenotyped by flow cytometry, and the SP was defined by efflux of fluorescent dye Hoechst 33342 visualized by ultraviolet excitation. Cells were flow sorted, and their colony-forming potential measured on feeder cells in culture. Results An SP was identified in islet and exocrine cells from human organ donors: 2 with type 1 diabetes, 3 with type 2 diabetes, and 28 without diabetes. Phenotyping revealed that exocrine SP cells had an epithelial origin, were enriched for carbohydrate antigen 19-9+ ductal cells expressing stem cell markers CD133 and CD26, and had greater colony-forming potential than non-SP cells. The exocrine SP was increased in a young adult with type 1 diabetes and ongoing islet autoimmunity. Conclusions The pancreatic exocrine SP is a potential reservoir of adult stem/progenitor cells, consistent with previous evidence that such cells are duct-derived and express CD133.
AB - Objectives The side population (SP) contains cells with stem cell/progenitor properties. Previously, we observed that the mouse pancreas SP expanded after pancreatic injury. We aimed to characterize the SP in human pancreas as a potential source of stem cells. Methods Human organ donor pancreata were fractionated into islets and exocrine tissue, enriched by tissue culture and dispersed into single cells. Cells were phenotyped by flow cytometry, and the SP was defined by efflux of fluorescent dye Hoechst 33342 visualized by ultraviolet excitation. Cells were flow sorted, and their colony-forming potential measured on feeder cells in culture. Results An SP was identified in islet and exocrine cells from human organ donors: 2 with type 1 diabetes, 3 with type 2 diabetes, and 28 without diabetes. Phenotyping revealed that exocrine SP cells had an epithelial origin, were enriched for carbohydrate antigen 19-9+ ductal cells expressing stem cell markers CD133 and CD26, and had greater colony-forming potential than non-SP cells. The exocrine SP was increased in a young adult with type 1 diabetes and ongoing islet autoimmunity. Conclusions The pancreatic exocrine SP is a potential reservoir of adult stem/progenitor cells, consistent with previous evidence that such cells are duct-derived and express CD133.
KW - CA 19-9 - carbohydrate antigen 19-9
KW - CFP - colony-forming potential
KW - diabetes
KW - DMEM - Dulbecco's modified Eagle medium
KW - FCS - fetal calf serum
KW - FSC - forward scatter
KW - GAD Ab - antibodies to the islet autoantigen glutamic acid decarboxylase (molecular weight, 65,000 dalton isoform)
KW - glutamic acid decarboxylase antibody
KW - HT - human tonicity
KW - human pancreas
KW - organ donor
KW - PI - propidium iodide
KW - RT - room temperature
KW - side population
KW - SP - side population
KW - SPc - side population cell
KW - SSC - side scatter
KW - stem cells
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U2 - 10.1097/MPA.0000000000000950
DO - 10.1097/MPA.0000000000000950
M3 - Article
C2 - 29135679
AN - SCOPUS:85039074537
SN - 0885-3177
VL - 47
SP - 25
EP - 34
JO - Pancreas
JF - Pancreas
IS - 1
ER -