title: A single-islet microplate assay to measure mouse and human islet insulin secretion.
abstract: One complication to comparing β-cell function among islet preparations, whether from genetically identical or diverse animals or human organ donors, is the number of islets required per assay. Islet numbers can be limiting, meaning that fewer conditions can be tested; other islet measurements must be excluded; or islets must be pooled from multiple animals/donors for each experiment. Furthermore, pooling islets negates the possibility of performing single-islet comparisons. Our aim was to validate a 96-well plate-based single islet insulin secretion assay that would be as robust as previously published methods to quantify glucose-stimulated insulin secretion from mouse and human islets. First, we tested our new assay using mouse islets, showing robust stimulation of insulin secretion 24 or 48 h after islet isolation. Next, we utilized the assay to quantify mouse islet function on an individual islet basis, measurements that would not be possible with the standard pooled islet assay methods. Next, we validated our new assay using human islets obtained from the Integrated Islet Distribution Program (IIDP). Human islets are known to have widely varying insulin secretion capacity, and using our new assay we reveal biologically relevant factors that are significantly correlated with human islet function, whether displayed as maximal insulin secretion response or fold-stimulation of insulin secretion. Overall, our results suggest this new microplate assay will be a useful tool for many laboratories, expert or not in islet techniques, to be able to precisely quantify islet insulin secretion from their models of interest.
authors.affiliation: a Department of Medicine ; Division of Endocrinology, Diabetes & Metabolism; University of Wisconsin-Madison ; Madison , WI USA.|c William S Middleton Memorial Veterans Hospital ; Madison , WI USA.
authors.forename: Nathan A
authors.initials: NA
authors.lastname: Truchan
authors.affiliation: a Department of Medicine ; Division of Endocrinology, Diabetes & Metabolism; University of Wisconsin-Madison ; Madison , WI USA.|c William S Middleton Memorial Veterans Hospital ; Madison , WI USA.
authors.forename: Harpreet K
authors.initials: HK
authors.lastname: Brar
authors.affiliation: a Department of Medicine ; Division of Endocrinology, Diabetes & Metabolism; University of Wisconsin-Madison ; Madison , WI USA.|c William S Middleton Memorial Veterans Hospital ; Madison , WI USA.
authors.forename: Shannon J
authors.initials: SJ
authors.lastname: Gallagher
authors.affiliation: b Interdisciplinary Graduate Program in Nutritional Sciences; University of Wisconsin-Madison ; Madison , WI USA.|c William S Middleton Memorial Veterans Hospital ; Madison , WI USA.
authors.forename: Joshua C
authors.initials: JC
authors.lastname: Neuman
authors.affiliation: a Department of Medicine ; Division of Endocrinology, Diabetes & Metabolism; University of Wisconsin-Madison ; Madison , WI USA.|b Interdisciplinary Graduate Program in Nutritional Sciences; University of Wisconsin-Madison ; Madison , WI USA.|c William S Middleton Memorial Veterans Hospital ; Madison , WI USA.
authors.forename: Michelle E
authors.initials: ME
authors.lastname: Kimple
mesh_terms: D000328:Adult; D000818:Animals; D001681:Biological Assay / Q000295:instrumentation / Q000379:methods*; D018929:Cell Culture Techniques / Q000295:instrumentation; D005260:Female; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology / Q000378:metabolism*; D008297:Male; D051379:Mice; D008875:Middle Aged; D055815:Young Adult
chemical_list: D007328:Insulin; D005947:Glucose
keywords: assay development; insulin secretion; islet culture; microplate assay; β-cell function