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Histopaque 提供最佳的小鼠胰岛纯化动力学:与 Ficoll、碘海醇和葡聚糖的比较研究。

Histopaque provides optimal mouse islet purification kinetics: comparison study with Ficoll, iodixanol and dextran.

机构信息

Department of Surgery, University of Alberta, Edmonton, Canada.

出版信息

Islets. 2011 Jul-Aug;3(4):144-9. doi: 10.4161/isl.3.4.15729. Epub 2011 Jul 1.

Abstract

Islet transplantation has become a very promising treatment for type 1 diabetes. To facilitate further clinical improvements in this exciting field, rodent islets are used to evaluate new strategies and modifications. One method to purify islets is on a density gradient, although the optimal gradient component can be debated. N=6 separate mouse islet isolations were used and the resulting islets were separated and purified on either a Ficoll, Histopaque, Dextran or Iodixanol gradient. Islets were assessed for recovery, viability, purity and in vitro functionality. Aliquots were transplanted into diabetic mice to assess in vivo functionality and survival. There was no difference in the number of islets recovered across groups nor in the size of recovered islets. Use of a Ficoll or Histopaque gradient led to the most pure and viable islets in comparison to Dextran and Iodixanol. Functionally, islets isolated on a Ficoll gradient had the highest glucose-stimulated insulin release in vitro while performing equally to Histopaque and Dextran gradients in vivo. Using a Ficoll gradient, however, comes at a higher monetary cost. We recommend using a Histopaque gradient, which led to the isolation of viable and functional islets with a reduced cost as compared to a Ficoll gradient.

摘要

胰岛移植已成为治疗 1 型糖尿病的一种极有前途的方法。为了促进这一令人兴奋的领域的进一步临床进展,啮齿动物胰岛被用于评估新的策略和改良方法。一种纯化胰岛的方法是基于密度梯度,但最佳的梯度成分可能存在争议。使用了 6 组单独的小鼠胰岛分离,然后将分离和纯化的胰岛分别置于 Ficoll、Histopaque、Dextran 或 Iodixanol 梯度上。评估胰岛的回收率、活力、纯度和体外功能。将胰岛移植到糖尿病小鼠中,以评估其体内功能和存活率。各组胰岛的回收率和回收胰岛的大小没有差异。与 Dextran 和 Iodixanol 相比,使用 Ficoll 或 Histopaque 梯度可获得最纯和最有活力的胰岛。在功能上,在 Ficoll 梯度上分离的胰岛在体外具有最高的葡萄糖刺激胰岛素释放,而在体内与 Histopaque 和 Dextran 梯度的功能相当。然而,使用 Ficoll 梯度的成本更高。我们建议使用 Histopaque 梯度,与 Ficoll 梯度相比,它可以分离出具有活力和功能的胰岛,同时降低成本。

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