title: A human islet cell culture system for high-throughput screening.
abstract: A small-molecule inducer of beta-cell proliferation in human islets represents a potential regeneration strategy for treating type 1 diabetes. However, the lack of suitable human beta cell lines makes such a discovery a challenge. Here, we adapted an islet cell culture system to high-throughput screening to identify such small molecules. We prepared microtiter plates containing extracellular matrix from a human bladder carcinoma cell line. Dissociated human islets were seeded onto these plates, cultured for up to 7 days, and assessed for proliferation by simultaneous Ki67 and C-peptide immunofluorescence. Importantly, this environment preserved beta-cell physiological function, as measured by glucose-stimulated insulin secretion. Adenoviral overexpression of cdk-6 and cyclin D(1), known inducers of human beta cell proliferation, was used as a positive control in our assay. This induction was inhibited by cotreatment with rapamycin, an immunosuppressant often used in islet transplantation. We then performed a pilot screen of 1280 compounds, observing some phenotypic effects on cells. This high-throughput human islet cell culture method can be used to assess various aspects of beta-cell biology on a relatively large number of compounds.
authors.affiliation: Chemical Biology Program, Broad Institute, Cambridge, MA 02142, USA.
authors.forename: Deepika
authors.initials: D
authors.lastname: Walpita
authors.forename: Thomas
authors.initials: T
authors.lastname: Hasaka
authors.forename: James
authors.initials: J
authors.lastname: Spoonamore
authors.forename: Amedeo
authors.initials: A
authors.lastname: Vetere
authors.forename: Karen K
authors.initials: KK
authors.lastname: Takane
authors.forename: Dina
authors.initials: D
authors.lastname: Fomina-Yadlin
authors.forename: Nathalie
authors.initials: N
authors.lastname: Fiaschi-Taesch
authors.forename: Alykhan
authors.initials: A
authors.lastname: Shamji
authors.forename: Paul A
authors.initials: PA
authors.lastname: Clemons
authors.forename: Andrew F
authors.initials: AF
authors.lastname: Stewart
authors.forename: Stuart L
authors.initials: SL
authors.lastname: Schreiber
authors.forename: Bridget K
authors.initials: BK
authors.lastname: Wagner
mesh_terms: D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D004353:Drug Evaluation, Preclinical; D005947:Glucose / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology* / Q000187:drug effects; D061251:Primary Cell Culture / Q000379:methods*; D015203:Reproducibility of Results; D054852:Small Molecule Libraries
chemical_list: D007328:Insulin; D054852:Small Molecule Libraries; D005947:Glucose