An Established Platform for Profiling Compounds for Diabetes
Until recently, cell based disease models to evaluate the effects of compounds to treat diabetes were limited to cell lines and rodent isolated islets. Our PHASEZERO™ Research team, pioneers in the development of human tissue based predictive disease models, has established the islet OrganDOT system. This three-dimensional (3D) tissue culture system models, under physiological and pharmacological control, the release of mediators from human islet cells, for example insulin from ß cells. The system provides actionable data on the effects of test compounds in a rapid and reproducible manner.
How the OrganDOT™ System Works
Freshly isolated and intact human pancreatic islets sourced from multi-organ transplant donors are dispersed into single cells and cultured on a membrane. Fed basolaterally with culture medium, over time the cells re-aggregate and retain islet-like functionality for 28 days. isletOrganDOTs can be challenged with physiological and pharmacological agents and secreted mediators measured.
GSIS assays are performed by first incubating each islet OrganDOT in the presence of 1.67mM glucose for 30 minutes to determine basal insulin secretion and then for 30 minutes in the presence of test solution (e.g. 6.7mM glucose ± test compound) to determine stimulated insulin secretion. Data are plotted as net insulin secretion (stimulated minus basal). Where the test solution is 1.67mM glucose, calculation of net insulin secretion yields a negative value for some donors.
Figure 1. Glucose stimulated insulin secretion. Data generated using isletOrganDOTs prepared from 10 different donors. Consistent stimulation of insulin secretion is measured in response to increased glucose concentrations. Data are mean ± sem, n=4 replicates.
Figure 2. Effect of secretagogues. Glucose-stimulated insulin secretion in islet OrganDOTs from a further 5 donors showing the effect of the standard secretagogue exendin-4. Data are mean ± sem, n= 4 replicates. Statistically significant potentiation of insulin secretion is shown (*, ***) compared to the respective glucose only treatment for each donor. islet OrganDOTs respond reproducibly to other standard secretagogues such as GLP-1 and sulphonylureas (data not shown).
KEY FEATURES: Retained Functionality, Higher Throughput, and Reliable Results
- isletOrganDOTs are human reconstituted primary islets that retain functionality for 28 days in culture, allowing for carefully timed and repeat treatments
- isletOrganDOTs are cultured on 24-membrane insert plates, enabling parallel testing of control and test agents
- Up to ten, 24-membrane insert plates can be run in one study, allowing for the flexible testing of batches of early
hit compounds to in-depth profiling of lead/pre-clinical compounds
- The small volume culture system minimizes
- Medium changes and testing regimes are applied basolaterally, reducing disruption to the isletOrganDOTs
Advantages of OrganDOT
- Robust design suitable for early hit to in-depth
lead/pre-clinical compound profiling- reducing the need to interpret results obtained from multiple assay platforms
- Simultaneous testing of multiple treatments and schedules delivers comprehensive compound profiles within a shorter time period
- Validated and quality controlled platform; each isletOrganDOT batch can be pre-screened to ensure experimental suitability
- Extensive background GSIS dataset generated in isletOrganDOT cultures from >50 individual donors