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URL: https://dx.doi.org/DOI:10.17504/protocols.io.bc3siyne

Authors: Integrated Islet Distribution Program

Group: Integrated Islet Distribution Program

Summary: The goal of the Integrated Islet Distribution Program (IIDP) is to develop a uniform standardized shipping method among all the subcontracted IIDP centers that will result in minimal loss of quality or quantity of shipped islets (as compared to control islets remaining at the IIDP center). The objective of this Standard Operating Procedure (SOP) is to be a model for site-specific SOPs that define the method for quantitative determination of insulin released after glucose stimulation for proving the potency of the human islet preparation shipped by the IIDP.GSIR Overview:After overnight culture at 37°C (12 – 24 hrs), human islets are incubated with media containing a relatively lowconcentration of glucose (2.8 mM) and a sample of the supernatant is taken. Then the same islets are incubatedwith media containing a higher glucose concentration (28 mM) and a sample of the supernatant is taken. Theamount of insulin present in both the supernatant samples is measured using a commercially available HumanInsulin ELISA kit. The stimulation index is calculated by dividing insulin concentration of the supernatantfrom the 28 mM glucose incubation by the insulin concentration of the supernatant from the 2.8 mM glucoseincubation.Reference:Purified Human Pancreatic Islets, Glucose Stimulated Insulin Release Determination by ELISA; DAIT, NIAID, NIH SOP Appendix, Document No. 3104, A03, Revision Number 03; Effective Date: 25 Oct 2010; Supersedes Date: 08 June 2010.

Affiliations: Integrated Islet Distribution Program, City of Hope, Duarte, CA

External URL: https://iidp.coh.org/Investigators/Policies-Standard-Operating-Procedures

Version: 1

Publication Date: 2020

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