Murine Model of Islet Transplantation: A Surgical Method to Infuse Pancreatic Islets in Portal Vein and Prevention of Postoperative Bleeding

Abstract

Source: Gou, W. et al. Minimizing Post-Infusion Portal Vein Bleeding during Intrahepatic Islet Transplantation in Mice. J. Vis. Exp. (2021)

In this video, we describe a procedure to infuse pancreatic islet cells into the hepatic portal vein, followed by application of gel foam to successfully prevent bleeding. Postoperative bleeding is a common cause of high mortality during islet transplantation.

Protocol

1. Islet transplantation Induce and maintain the mouse under general anesthesia with 2% isoflurane. Check for the lack of pedal reflexes to ensure proper anesthetization of the animal. Shave and remove the fur in the abdomen area of the mouse. Administer a single pre-operative dose of Buprenorphine (0.1 mg/kg i.p.). Disinfect the surgical area with three alternating wipes of 2% iodine and 75% alcohol. Perform a laparotomy with micro scissors to generate…

Representative Results

Figure 1: Illustration of intrahepatic islet transplantation procedures. (A, C). Schematics of key steps used in Method A and Method B. (B, D). Islets were injected directly via the portal vein (C) or indirectly via fat pat (D).

Materials

10% Neutral buffered formalin v/v	Fisher Scientific	23426796
1 mL Syringe with needle	AHS	AH01T
20 mL Syringe	BD	301031
25G x 5/8" hypodermic needles	BD	305122
Alcohol prep pads, sterile	Fisher Scientific	22-363-750
Animal Anesthesia system	VetEquip, Inc.	Inc. 901806
Buprenorphine hydrochloride, injection	Par Sterile Products LLC	NDC 42023-179-05
Ethanol absolute (200 proof), molecular biology grade	Fisher Scientific	BP2818500
Extra fine Micro Dissecting scissors 4” straight sharp	Roboz Surgical Instrument Co.	RS-5882
Gelfoam (absorbable gelatin sponge USP)	Pharmacia & Upjohn Company	34201
Graefe forceps 4” extra delicate tip	Roboz Surgical Instrument Co.	RS-5136
Heated pad	Amazon	B07HMKMBKM
Hegar-Baumgartner Needle Holder 5.25”	Roboz Surgical Instrument Co.	RS-7850
Insulin syringe with 27-gauge needle	BD	879588
Iodine prep pads	Fisher Scientific	19-027048
Isoflurane	Piramal Critical Care	NDC 66794-017-25
Polypropylene Suture 4-0	Med-Vet International	MV-8683
Polypropylene Suture 5-0	Med-Vet International	MV-8661
Sodium chloride,  0.9% intravenous solution	VWR	2B1322Q
Surgical drape, sterile	Med-Vet International	DR1826