ABSTRACT
The aim of this study was to investigate the effect of culture at 24 degrees C on cell viability, cellular function, immunogenicity, and cytokine profiles of rat pancreatic islets. Pancreatic islets were isolated from Lewis rats and cultured at either 24 degrees C or 37 degrees C for 14 days. Islet recovery was counted as islet equivalents; islet viability was examined with fluorescent vital staining. Islet function was measured with a glucose stimulation test. Annexin V, and MHC class I and II expression were measured using flow cytometric assay for apoptosis and immunogenicity, respectively. Lymphocyte cell proliferation was examined with WST-1 proliferation assay. Cytokine profiles were analyzed with quantitative real time RT-PCR. All these parameters were measured on 1, 3, 5, 7 and 14 culture days after islet isolation. Islet recovery was higher in islets cultured at 24 degrees C than 37 degrees C without a change in viability. Insulin secretion after glucose stimulation was more effective in 24 degrees C culture conditions. Decreased apoptotic cell death was demonstrated in 24 degrees C cultured islets. Both MHC class I and II expression on islets and lymphocyte proliferation upon coculture with islets were less prominent in 24 degrees C cultured islets. TNF-alpha expression was lower in islets cultured at 24 degrees C than in islets cultured at 37 degrees C. Both IL-1beta and IL-10 cytokine expressions were similar under both culture conditions. This study demonstrated that cell recovery and function are increased in islets cultured at 24 degrees C than those at 37 degrees C with decreased antigenicity and proinflammatory cytokine expression.
