Here is a good protocol to do that : http://link.springer.com/protocol/10.1385/0-89603-335-X%3A391

Islets are microorgans (Soria 2000 Engineeering pancreatic islets Eur J Physiol) and you may have difficulties in keeping them in culture because oxygen does not reach the central part of a 200 um islet.

1. First disperse the islets, gentle aspiration with micropipete, or dissociate cells with low calcium (not too long, less than 5-10 min) with gentle aspiration.

2. Form agregates of 10-15 cells (increasing glucose may work)

3. culture them like other cells, but primary culture does not last more than 1 week (aprox)

good luck

Abraham,

We have similar experience to what is mentioned by Bernat. Without breaking up islets into smaller cell clusters, you will lose a substantial portion of central cells in islets that are 100µm or larger in diameter in less than 3 days. Smaller islets (about 50µm diameter) survive. However, as the time passes, their behavior (insulin secretory responses to normal secretogogues) diminishes. For some of these reasons, when it was necessary to use isolated islets, we have always tried to finish experiments in under 3 days. But mostly, we have either used in vivo models or used isolated pancreatic perfusions.

Best...