Establishment Of A Culture System For Rabbit Pancreatic Islet Cells In Vitro And Effect Of Auricularla Auricula Polysaccharides On The Celluar Activity

Currently, the isolating, purifying and in vitro culture of SD rat pancreatic cells is more developed among in vitro culture systems of pancreatic cells. However, some of the rabbit gene sequence is highly homologous to those of human. Therefore, establishing a in vitro culture system of rabbit pancreatic cells is of great significance.In the current work, isolation and purification of islet cells were achieved by differential adhesion method which was based on the difference of adhesion speed rates between islet cells and fibroblasts after collagenase digestion of pancreatic tissue. The isolated islet cells were identified by specific DTZ stain. Afterward, RPMI-1640medium, M199medium, DMEM low glucose medium, DMEM high glucose culture medium were chosen as candidates of optimal medium. After five days’culturing, cell activitiy of islet cells developed in the four media above was evaluated by MTT method. The results suggested cells in RPMI-1640medium bore higher activity, so RPMI-1640was the optimal medium for rabbit islet cells. Since culture medium was decided, serum concentration of the medium was optimized by adjusting the concentration to20%,15%,10%and5%respectively. Also, after five days’ culturing, cell activity in different concentration was determined by MTT. The results indicated20%was optimal. In the present research, in vitro rabbit islet cells culture was conducted, adding Auricularia auricular polysaccharide at concentrations of400mg/L and800mg/L to the media. Cell activity was evaluated by MTT, insulin secretion under low and high saccharide levels was determined by ELISA and expression levels of anti-apoptosis gene bcl-2of the pancreatic islet cells were determined by RT-PCR. Results showed Auricularia auricular polysaccharide had no significant effect on cell activity, insulin secretion and proliferation at concentrations of400mg/L and800mg/L.