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建立猫胰岛分离的方案。

Establishment of a protocol for the isolation of feline pancreatic islets.

机构信息

Institute of Veterinary Physiology, Vetsuisse Faculty University of Zurich, Winterthurerstrasse 260, CH-8057 Zurich, Switzerland.

Clinic for Small Animal Internal Medicine, Vetsuisse Faculty, University of Zurich, Winterthurerstrasse 260, CH-8057 Zurich, Switzerland.

出版信息

Physiol Behav. 2018 Mar 15;186:79-81. doi: 10.1016/j.physbeh.2018.01.012. Epub 2018 Jan 31.

Abstract

Diabetes mellitus is a common endocrinopathy in cats that is associated with pancreatic islets lesions. Research on isolated islets contributed to the understanding of the pathophysiology of human diabetes. Therefore, by improving the existing methods of isolation in cats, we aimed at increasing islet yield, purity and viability of feline isolated islets. Islet isolation was accomplished by pancreas perfusion with 80ml of Collagenase type IV through the pancreatic duct at the site of the major papilla. The enzymatic digestion was combined with mechanical disruption and controlled by dithizone staining. Purification was performed by filtration and handpicking. Purified islets were plated on extracellular matrix pre-coated plates and cultured for 48h. Feline islets with a high degree of viability and purity were isolated and cultured for the first time. Although the percentage of islet free from the acinar tissue relative to the total number of isolated islets was low compared to other species, the suggested protocol represents a promising progress in the procedure of islet isolation in cats.

摘要

糖尿病是一种常见的猫内分泌疾病,与胰岛损伤有关。对分离胰岛的研究有助于了解人类糖尿病的病理生理学。因此,通过改进猫胰岛的现有分离方法,我们旨在提高胰岛的产量、纯度和活力。胰岛分离通过在主乳头处的胰管内灌注 80ml IV 型胶原酶来完成。酶消化与机械破坏相结合,并通过二硫代丁二酸钠染色进行控制。纯化通过过滤和手工挑选进行。纯化的胰岛接种在预先涂有细胞外基质的平板上,并培养 48 小时。首次成功分离和培养了具有高活力和纯度的猫胰岛。尽管与其他物种相比,相对于分离的胰岛总数,无腺泡组织的胰岛比例较低,但所提出的方案代表了猫胰岛分离程序的一个有希望的进展。

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