isolation of human umbilical vein endothelial cells using kiwifruit actinidin

introduction: proteolytic enzymes, specially collagenase, are used to digest extracellular matrix, cells isolation and primary culture. it is important to find new sources of plant or animal protease instead of bacterial or tissue collagenase. in the present research, actinidin, a plentiful protease in kiwifriut, was used to isolate human umbilical vein endothelial cells. materials and methods: human umbilical vein endothelial cells (huvec) were isolated using different doses of actinidin (from 1 to 16 mg/ml) and different incubation times (from 10 to 60 minutes). freshly isolated endothelial cells were cultured in mcdb131 medium. endothelial cells were identified by their non-overlapping cobblestone morphology and immunostaining. the viability of separated cells was assessed by trypan blue exclusion test. results: actinidin in concentration of 4 mg/ml for 20-30 minutes selectively isolates human umbilical vein endothelial cells with minimal contamination of other cell populations. the viability of separated cells was estimated 90-95% in this situation. conclusion: the results showed that actinidin has not toxic effect on separated cells and is a novel and suitable protease for isolation of huvec cells.