Effects of different methods of isolating and purifying rat islet cells on their loss

Objective: To explore the best method of isolating and purifying rat islet cells to improve the harvest and purity of rat islet cells. Methods: Forty rats were divided into four groups, with ten rats per group. Group A: Rat pancreases were perfused with Hanks and digested by collagens V, islet cells were isolated and purified by gradient Ficoll400. Group B: Pancreases were cut and ground, then filtered with grid. Group C: Rat Pancreases were perfused with collagens V, islet cells were isolated and purified by gradient Ficoll400. Group D: Rat pancreases were perfused with collagens V, but filtered by grid. The harvest and purity of islet cells from four groups were compared and analysed. Results: The differences in the harvest and purity of islet cells isolated and purified by the four methods (A, B, C and D groups) were significant by completely random variance analysis. Interclass multiple comparison manifested that the harvest in A group was higher than that in Group B and D, but not obviously different from that in Group C. The purity in A group was not significantly different from that in other groups. Conclusion: The harvest and purity of islet cells isolated through Hanks perfusion and/or collagenase V digestion were higher than those of other methods. The cost of this method is lower, so it suggests that it is a better method for islet cell isolation.