Pancreatic islet culture for islet transplantation

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Islet transplantation is a novel diabetic therapy instead of insulin injection. In vitro culture of islet is pivotal process, because islet cells sometimes lose their function during in vitro culture. In this study, we focused on matrices coating so as to improve the culture and prolong their insulin secretion. In order to investigate the effect of matrices, the islet cells were cultured on dishes coated with several matrices, and measured the detachment of the cells from them and insulin release in response to glucose level.The islets were isolated from male Lewis rats and then dispersed with trypsin. Dispersed cells were seeded on the dishes coated with type I collagen, type IV collagen, fibronectin, laminin or without. Intensity of detachment of the cells to the matrices was assayed by counting the detached cell number after 20 times pipetting. Glucose stimulation test was carried out in glucose-rich medium and glucose-poor medium, and the insulin amount in the culture supernatant was determined by ELISA. The islet cells cultured on type I collagen and laminin were most easily detached after pipetting, and cells on type IV collagen were also easily detached, but cells on fibronectin and noncoated plastic were failed to detach. Cells cultured on laminin released most amount of insulin and showed highest stimulation index, but the cells on fibronectin or noncoated plastic released the least insulin and showed the lowest stimulation index. These results imply that attachment on the culture dish could inhibit insulin release from islets cells and that islet cells should be cultured on easily detachable dish.