A single-islet microplate assay to measure mouse and human islet insulin secretion. Islets 2015;7(3):e1076607
Date
10/11/2015Pubmed ID
26452321Pubmed Central ID
PMC4708880DOI
10.1080/19382014.2015.1076607Scopus ID
2-s2.0-84964066497 (requires institutional sign-in at Scopus site) 25 CitationsAbstract
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.
Author List
Truchan NA, Brar HK, Gallagher SJ, Neuman JC, Kimple MEAuthor
Nathan A. Truchan PhD Research Scientist I in the Pediatrics department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AdultAnimals
Biological Assay
Cell Culture Techniques
Female
Glucose
Humans
Insulin
Islets of Langerhans
Male
Mice
Middle Aged
Young Adult
