Chadwick D R, Robertson G S, Contractor H, Swift S, Rose S, Thirdborough S T, Chamberlain R, James R F, Bell P R, London N J
Department of Surgery, Leicester Royal Infirmary, UK.
Acta Diabetol. 1993;30(1):57-9. doi: 10.1007/BF00572876.
Euro-Ficoll (EF) and bovine serum albumin (BSA) are the two most commonly used media for the density gradient purification of human pancreatic islets. The aim of this study was to compare these two media with respect to the efficiency of human islet isolation. Ten human pancreata were collagenase-digested, and samples of digest were separated on either a continuous linear density gradient of BSA or a discontinuous gradient of EF (1.108/1.096/1.037/Euro-Collins). Efficiency of islet purification was assessed by insulin and amylase assay of aliquots aspirated from the BSA gradients, and from the interfaces of the EF gradients. Islets were obtained from two interfaces in the EF gradients. Islet yield from the upper interface was generally poor (median 28% of total insulin; range 2-71%), but purity was better than for an equivalent yield using BSA [1% (0-3%) amylase contamination for EF versus 6% (0-37%) for BSA; P = 0.013]. Pooling both EF interfaces increased yield to 66% (17-81%) but markedly reduced purity [46% (0-50%) amylase for EF versus 31% (0-52%) for BSA]. In conclusion, the efficiency of human islet purification is similar, though disappointingly low, with BSA and with EF. Considerable scope exists, therefore, for improvement in the density gradient purification of human islets.
欧洲菲可（EF）和牛血清白蛋白（BSA）是用于人胰岛密度梯度纯化的两种最常用介质。本研究旨在比较这两种介质在人胰岛分离效率方面的差异。对10个人类胰腺进行胶原酶消化，消化样本在BSA连续线性密度梯度或EF间断梯度（1.108/1.096/1.037/欧洲柯林斯液）上进行分离。通过对从BSA梯度以及EF梯度界面吸出的等分试样进行胰岛素和淀粉酶测定来评估胰岛纯化效率。从EF梯度的两个界面获得胰岛。上界面的胰岛产量通常较低（胰岛素总量中位数为28%；范围为2 - 71%），但纯度优于使用BSA获得的同等产量时的纯度[EF的淀粉酶污染为1%（0 - 3%），而BSA为6%（0 - 37%）；P = 0.013]。将EF的两个界面的胰岛合并可使产量提高到66%（17 - 81%），但显著降低了纯度[EF的淀粉酶含量为46%（0 - 50%），而BSA为31%（0 - 52%）]。总之，使用BSA和EF进行人胰岛纯化的效率相似，尽管令人失望地低。因此，在人胰岛密度梯度纯化方面仍有很大的改进空间。
