An optimized mixture of kiwifruit actinidin and trypsin for isolation and culture of endothelial cells from rat aorta

Proteolytic enzymes, especially collagenases, are used for digestion of extracellular matrix, cell isolation and primary culture. Because of the problems in purification and low amount of collagenases in bacterial or animal sources, it is important to find new sources of the enzymes. So, in the present study actinidin, a plentiful protein in kiwifruit was purified and a mixture of actinidin and trypsin was applied to isolate rat aortic endothelial cells. Aortic endothelial cells were isolated using digestion solution containing different concentrations of actinidin (from 2 to 16 mg/ml) and trypsin