title: A novel dual-color reporter for identifying insulin-producing beta-cells and classifying heterogeneity of insulinoma cell lines.
abstract: 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.
authors.affiliation: Department of Physiology & Biophysics and Zilkha Neurogenetics Institute, University of Southern California, Los Angeles, California, United States of America. nanlee@usc.edu
authors.forename: Nan Sook
authors.initials: NS
authors.lastname: Lee
authors.forename: Joyce G
authors.initials: JG
authors.lastname: Rohan
authors.forename: Madison
authors.initials: M
authors.lastname: Zitting
authors.forename: Sonia
authors.initials: S
authors.lastname: Kamath
authors.forename: Andrew
authors.initials: A
authors.lastname: Weitz
authors.forename: Arnold
authors.initials: A
authors.lastname: Sipos
authors.forename: Paul M
authors.initials: PM
authors.lastname: Salvaterra
authors.forename: Kouichi
authors.initials: K
authors.lastname: Hasegawa
authors.forename: Martin
authors.initials: M
authors.lastname: Pera
authors.forename: Robert H
authors.initials: RH
authors.lastname: Chow
mesh_terms: D002118:Calcium / Q000378:metabolism; D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic; D023061:Gene Order; D017930:Genes, Reporter; D005822:Genetic Vectors; D006801:Humans; D007328:Insulin / Q000235:genetics; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D007340:Insulinoma; D015640:Ion Channel Gating; D010641:Phenotype; D011401:Promoter Regions, Genetic
chemical_list: D007328:Insulin; D002118:Calcium