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牛血清白蛋白密度梯度法分离大鼠胰岛。

Bovine serum albumin density gradient isolation of rat pancreatic islets.

作者信息

Lake S P, Anderson J, Chamberlain J, Gardner S J, Bell P R, James R F

出版信息

Transplantation. 1987 Jun;43(6):805-8.

PMID:3109087
Abstract

The use of a bovine serum albumin (BSA) density gradient for isolation of rat pancreatic Islets of Langerhans after collagenase digestion has been compared with the standard Ficoll separation technique. The criteria studied were islet yield (insulin extraction of the islet interfaces and pellet), purity of preparation (amylase content of the islet preparation), insulin release characteristics, and the result of isologous transplantation in diabetic rats. The islet interface of the BSA gradient contained 62.2% of the total insulin, whereas the corresponding interface of the Ficoll gradient contained only 36.6% (P less than 0.001). The amylase content of the Ficoll-separated islet preparation was 6133 U/L, as opposed to 1230 U/L with BSA (P less than 0.001). BSA-isolated islets gave similar insulin release characteristics to non-density-gradient-isolated islets, whereas Ficoll-separated islets showed suboptimal insulin release. Single-donor-single-recipient transplantation was successfully performed with BSA-isolated islets whereas multiple donors were required with Ficoll-separated islets. Thus significantly improved results were found with the bovine serum albumin density gradient separation in all criteria and consequently the use of this gradient represents an advance in islet isolation techniques.

摘要

已将牛血清白蛋白(BSA)密度梯度用于在胶原酶消化后分离大鼠胰岛,并与标准的聚蔗糖分离技术进行了比较。所研究的标准包括胰岛产量(胰岛界面和沉淀中的胰岛素提取量)、制剂纯度(胰岛制剂中的淀粉酶含量)、胰岛素释放特性以及在糖尿病大鼠中进行同种移植的结果。BSA梯度的胰岛界面含有总胰岛素的62.2%,而聚蔗糖梯度的相应界面仅含有36.6%(P<0.001)。聚蔗糖分离的胰岛制剂中的淀粉酶含量为6133 U/L,而BSA分离的为1230 U/L(P<0.001)。BSA分离的胰岛具有与非密度梯度分离的胰岛相似的胰岛素释放特性,而聚蔗糖分离的胰岛则表现出次优的胰岛素释放。使用BSA分离的胰岛成功进行了单供体-单受体移植,而聚蔗糖分离的胰岛则需要多个供体。因此,在所有标准方面,牛血清白蛋白密度梯度分离均取得了显著改善的结果,因此使用这种梯度代表了胰岛分离技术的一项进步。

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