Isolation, purification and functional identification of Syrian golden hamster islets / 南方医科大学学报

ABSTRACT

<p><b>OBJECTIVE</b>To establish the methods for isolation, purification and function identification of Syrian golden hamster islets.</p><p><b>METHODS</b>The Langerhans islets were isolated and purified from golden hamster pancreas by intra-ductal collagenase V perfusion and discontinuous Ficoll density gradient centrifugation. After isolation, the islet yield and purity were evaluated with DTZ staining. The islet function was assessed using glucose-stimulated insulin secretion test.</p><p><b>RESULTS</b>The total number of purified islets from one donor hamster was 359-/+35 islet equivalent (IEQ), with the purity and viability of the isolated islets of more than 90%. In response to glucose stimulations at 5.8 and 16.7 mmol/L, the secretion of insulin by the islets was 3.29-/+0.3 and 11.12-/+0.57 mU/L, respectively, showing a 2.28-fold higher insulin release by high-concentration than by low- concentration glucose stimulation (P<0.01).</p><p><b>CONCLUSION</b>The methods of collagenase V digestion and gradient centrifugation result in high yield and high purity of the isolated hamster islets.</p>