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小鼠胰岛的腹股沟皮下白色脂肪组织(ISWAT)移植模型

Inguinal Subcutaneous White Adipose Tissue (ISWAT) Transplantation Model of Murine Islets.

作者信息

Peng Yuanzheng, Zou Zhicheng, Chen Jiao, Zhang Hancheng, Lu Ying, Bittino Rito, Fu Hongxing, Cooper David K C, Lin Shan, Cao Mengtao, Dai Yifan, Cai Zhiming, Mou Lisha

机构信息

Shenzhen Xenotransplantation Medical Engineering Research and Development Center, Institute of Translational Medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Health Science Center.

Institute for Cellular Therapeutics, Allegheny-Singer Research Institute.

出版信息

J Vis Exp. 2020 Feb 16(156). doi: 10.3791/60679.

Abstract

Pancreatic islet transplantation is a well-established therapeutic treatment for type 1 diabetes. The kidney capsule is the most commonly used site for islet transplantation in rodent models. However, the tight kidney capsule limits the transplantation of sufficient islets in large animals and humans. The inguinal subcutaneous white adipose tissue (ISWAT), a new subcutaneous space, was found to be a potentially valuable site for islet transplantation. This site has better blood supply than other subcutaneous spaces. Moreover, the ISWAT accommodates a larger islet mass than the kidney capsule, and transplantation into it is simple. This manuscript describes the procedure of mouse islet isolation and transplantation in the ISWAT site of syngeneic diabetic mouse recipients. Using this protocol, murine pancreatic islets were isolated by standard collagenase digestion and a basement membrane matrix hydrogel was used for fixing the purified islets in the ISWAT site. The blood glucose levels of the recipient mice were monitored for more than 100 days. Islet grafts were retrieved at day 100 after transplantation for histological analysis. The protocol for islet transplantation in the ISWAT site described in this manuscript is simple and effective.

摘要

胰岛移植是1型糖尿病一种成熟的治疗方法。肾包膜是啮齿动物模型中最常用的胰岛移植部位。然而,紧密的肾包膜限制了在大型动物和人类中移植足够数量的胰岛。腹股沟皮下白色脂肪组织(ISWAT)作为一个新的皮下空间,被发现是胰岛移植一个潜在的有价值的部位。该部位的血液供应比其他皮下空间更好。此外,ISWAT比肾包膜能容纳更大的胰岛团块,且移植操作简单。本手稿描述了在同基因糖尿病小鼠受体的ISWAT部位进行小鼠胰岛分离和移植的过程。按照此方案,通过标准胶原酶消化法分离小鼠胰岛,并使用基底膜基质水凝胶将纯化后的胰岛固定于ISWAT部位。对受体小鼠的血糖水平进行了100多天的监测。在移植后第100天取出胰岛移植物进行组织学分析。本手稿中描述的在ISWAT部位进行胰岛移植的方案简单且有效。

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