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目的评价Dextran不连续密度梯度离心法纯化大鼠胰岛的效果。方法采用V型胶原酶分离出大鼠胰岛,并应用Dextran不连续密度梯度离心法纯化胰岛。在体视镜下计数双硫腙染色的胰岛并测量染色胰岛的直径。放免法测定胰岛素含量。AO-PI双染色确定胰岛的活力。结果平均每只成年Wistar大鼠的胰腺可分离950±24个胰岛,经Dextran纯化后平均每只成年Wistar大鼠的胰腺可获得784±10个胰岛,纯度可达到90%以上。结论采用Dextran不连续密度梯度纯化得到的Wistar大鼠胰岛结构完整、功能良好。
Objective To evaluate the effect of Dextran discontinuous density gradient centrifugation on rat islets. Methods Rat islets were isolated by collagenase type V and islets were purified using Dextran discontinuous density gradient centrifugation. Dithizone-dyed islets were counted under a stereo microscope and the diameter of the dyed islets was measured. RIA assay insulin content. AO-PI double staining to determine islet activity. Results On average, 950 ± 24 islets were isolated from the pancreas of each adult Wistar rat. After Dextran purification, an average of 784 ± 10 islets per pancreas of adult Wistar rats were obtained with a purity of over 90%. Conclusion Wistar rats purified by Dextran discontinuous density gradient have complete structure and good function.