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. 2018 Mar 5;8(5):e2743.
doi: 10.21769/BioProtoc.2743.

Murine Pancreatic Islets Transplantation under the Kidney Capsule

Tatiana Jofra  1 , Giuseppe Galvani  1 , Fousteri Georgia  1 , Gregori Silvia  2 , Nicola Gagliani  3   4 , Manuela Battaglia  1
Affiliations

Murine Pancreatic Islets Transplantation under the Kidney Capsule

Tatiana Jofra et al. Bio Protoc. .

Abstract

Type 1 diabetes (T1D) is an autoimmune disease caused by the lack of insulin-producing pancreatic beta cells leading to systemic hyperglycemia. Pancreatic islet transplantation is a valid therapeutic approach to restore insulin loss and to promote adequate glycemic control. Pancreatic islet transplantation in mice is an optimal preclinical model to identify new therapeutic strategies aiming at preventing rejection and optimizing post-transplant immuno-suppressive/-tolerogenic therapies. Islet transplantation in preclinical animal models can be performed in different sites such the kidney capsule, spleen, bone marrow and pancreas. This protocol describes murine islet transplantation under the kidney capsule. This is a widely accepted procedure for research purposes. Stress caused in the animals is minimal and it leads to reliable and reproducible results.

Keywords: Islet transplantation; Murine model; Pancreatic islets; Type 1 diabetes.

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Figures

Figure 1.
Figure 1.. Images of pancreatic islets.
Islets isolated before hand-picking (A). Purified islets after hand-picking (B). Scale bars represent 150 µm.
Figure 2.
Figure 2.. Collection and preparation of pancreatic islets.
Healthy pancreatic islets transferred into a Petri dish are collected with a P200 (A) and transferred into a sterile 1.5 ml Eppendorf tube (B).
Figure 3.
Figure 3.. Collection of pancreatic islets with Hamilton syringe.
Pancreatic islets are collected with the Hamilton syringe into a P200 tip (A) and then moved into the PE 50 tube (B).
Video 1.
Video 1.. Preparation of islets for transplantation.
Pancreatic islets are collected into a P200 tip and transferred into the PE50 tubing by slowly turned the Hamilton syringe.
Video 2.
Video 2.. Mouse preparation for islet transplantation.
The transplant area is disinfected and the mouse’s eyes are kept humid.
Video 3.
Video 3.. Incision and kidney exposure.
A small incision is made in the skin and in the peritoneum exposing the kidney.
Figure 4.
Figure 4.. Islets under the kidney capsule.
Islets (white) are localized under the kidney capsule. Scale bar represent 3 mm.
Video 4.
Video 4.. Islet transplantation.
Pancreatic islets are transplanted under the kidney capsule.
Figure 5.
Figure 5.. Surgical seams
Video 5.
Video 5.. Closing and mouse revival.
The peritoneum and the skin are sutured. Subcutaneous injection of ketoprofen is made and the mice are placed on a heating pad until are active.
See this image and copyright information in PMC

References

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