Functional proteomics screen enables enrichment of distinct cell types from human pancreatic islets

The current world-wide epidemic of diabetes has prompted attempts to generate new sources of insulin-producing cells for cell replacement therapy. An inherent challenge in many of these strategies is the lack of cell-surface markers permitting isolation and characterization of specific cell types fr...

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Bibliographic Details
Published inPloS one Vol. 10; no. 2; p. e0115100
Main Authors Sharivkin, Revital, Walker, Michael D, Soen, Yoav
Format Journal Article
LanguageEnglish
Published United States Public Library of Science 23.02.2015
Public Library of Science (PLoS)
Subjects
Acinar cells
Acinar Cells - cytology
Acinar Cells - metabolism
Arrays
Beta cells
Biological properties
Biological samples
Biomarkers - metabolism
CD56 antigen
CD9 antigen
Cell surface
Diabetes
Diabetes mellitus
Diabetes therapy
Diagnostic software
Diagnostic systems
Epidemics
Flow Cytometry
Gene expression
Glucagon
Glucagon-Secreting Cells - cytology
Glucagon-Secreting Cells - metabolism
Humans
Identification
Immunoglobulins
Insulin
Insulin-Secreting Cells - cytology
Insulin-Secreting Cells - metabolism
Islets of Langerhans
Islets of Langerhans - cytology
Islets of Langerhans - metabolism
Labeling
Markers
Pancreas
Proteins
Proteomics
Proteomics - methods
Purification
Rodents
Stem cells
Surface markers
Trypsin
Israel
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Summary:The current world-wide epidemic of diabetes has prompted attempts to generate new sources of insulin-producing cells for cell replacement therapy. An inherent challenge in many of these strategies is the lack of cell-surface markers permitting isolation and characterization of specific cell types from differentiating stem cell populations. Here we introduce an iterative proteomics procedure allowing tag-free isolation of cell types based on their function. Our method detects and associates specific cell-surface markers with particular cell functionality by coupling cell capture on antibody arrays with immunofluorescent labeling. Using this approach in an iterative manner, we discovered marker combinations capable of enriching for discrete pancreatic cell subtypes from human islets of Langerhans: insulin-producing beta cells (CD9high/CD56+), glucagon-producing alpha cells (CD9-/CD56+) and trypsin-producing acinar cells (CD9-/CD56-). This strategy may assist future beta cell research and the development of diagnostic tools for diabetes. It can also be applied more generally for function-based purification of desired cell types from other limited and heterogeneous biological samples.
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Conceived and designed the experiments: RS MDW YS. Performed the experiments: RS. Analyzed the data: RS MDW YS. Contributed reagents/materials/analysis tools: MDW YS. Wrote the paper: RS MDW YS.
Competing Interests: The authors have read the journal’s policy, and the authors of this manuscript have the following competing interests: 1. An International PCT patent application (no: WO14/030166 A1) has been filed by the Weizmann Institute. It is entitled: ‘METHODS OF ISOLATING DISTINCT PANCREATIC CELL TYPES’. 2. This study was supported by: (1) a grant (to Y.S. and M.W.) from the Juvenile Diabetes Research Foundation (JDRF) and the Israel Science Foundation (ISF), and (2) the Leona M. and Harry B. Helmsley Charitable Trust (to Y.S.). This does not alter the authors’ adherence to all PLOS ONE policies on sharing data and materials.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0115100