大鼠不同分离纯化方法对胰岛细胞丢失的影响

目的探讨提高大鼠胰岛细胞产量和纯度的最佳分离纯化方法。方法40只大鼠分为4组,每组10只。A组:Hanks液灌注 胶原酶V消化 Ficoll400梯度纯化;B组:组织剪碎研磨 筛网过滤;C组:胶原酶V灌注 Ficoll400梯度纯化;D组:胶原酶V灌注 筛网过滤。对用四种不同方法分离纯化的结果进行比较。结果完全随机设计的方差分析显示,A、B、C、D组收获量及纯度比较均有显著性差异(P<0.05);组间多重比较显示,A组收获量均高于B、D组(P<0.05),与C组比较差异无统计学意义(P>0.05);A组纯度与其他组比较均无显著性差异(P>0.05)。结论用胶原酶V或Hanks液灌注,Ficoll400梯度纯化法所收获胰岛细胞的产量和纯度较高,而且价格低廉,是一种较理想的胰岛细胞分离纯化方法。

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.