A novel dual-color reporter for identifying insulin-producing beta-cells and classifying heterogeneity of insulinoma cell lines

Many research studies use immortalized cell lines as surrogates for primary beta- cells. We describe the production and use of a novel "indirect" dual-fluorescent reporter system that leads to mutually exclusive expression of EGFP in insulin-producing (INS(+)) beta-cells or mCherry in non-...

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Bibliographic Details
Published in:PloS one 2012-04, Vol.7 (4), p.e35521-e35521
Main Authors: Lee, Nan Sook, Rohan, Joyce G, Zitting, Madison, Kamath, Sonia, Weitz, Andrew, Sipos, Arnold, Salvaterra, Paul M, Hasegawa, Kouichi, Pera, Martin, Chow, Robert H
Format: Article
Language:English
Subjects:
Adipocytes
Analysis
Beta cells
Biology
Biomedical engineering
Biophysics
Biotechnology
Calcium - metabolism
Cell Line, Tumor
Cell lines
Cloning
Color
Efficiency
Flow cytometry
Fluorescence
Gene expression
Gene Expression Regulation, Neoplastic
Gene Order
Genes, Reporter
Genetic Vectors
Glucose
Humans
Insulin
Insulin - genetics
Insulin-Secreting Cells - metabolism
Insulinoma
Ion Channel Gating
Journalists
Medical research
Medicine
Metabolism
Neuroendocrine tumors
Neurosciences
Phenotype
Physiology
Pluripotency
Promoter Regions, Genetic
Proteins
Purification
Rodents
Stem cells
Vectors (Biology)
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Summary:Many research studies use immortalized cell lines as surrogates for primary beta- cells. We describe the production and use of a novel "indirect" dual-fluorescent reporter system that leads to mutually exclusive expression of EGFP in insulin-producing (INS(+)) beta-cells or mCherry in non-beta-cells. Our system uses the human insulin promoter to initiate a Cre-mediated shift in reporter color within a single transgene construct and is useful for FACS selection of cells from single cultures for further analysis. Application of our reporter to presumably clonal HIT-T15 insulinoma cells, as well as other presumably clonal lines, indicates that these cultures are in fact heterogeneous with respect to INS(+) phenotype. Our strategy could be easily applied to other cell- or tissue-specific promoters. We anticipate its utility for FACS purification of INS(+) and glucose-responsive beta-like-cells from primary human islet cell isolates or in vitro differentiated pluripotent stem cells.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0035521