Antegrade Perfusion Model: An Ex Vivo Technique to Perfuse Isolated Mouse Heart

Abstract

Source: Omatsu-Kanbe, M. et al., An Antegrade Perfusion Method for Cardiomyocyte Isolation from Mice. J. Vis. Exp. (2021).

This video describes an antegrade perfusion technique of an isolated mouse heart during which a perfusate is injected through the left ventricle, and eventually, it perfuses the entire myocardium. This method can be used to isolate cardiomyocytes from murine models and generate decellularized hearts for further downstream applications.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Antegrade perfusion of the mouse heart

NOTE: The plastic transfer pipette used for sucking the heart should be soft and not be sharply tapered towards the tip. Choose a small vascular clamp with serration. The recommended instruments are listed in the Table of Materials.

1. Excision of the mouse heart and clamping the aorta
   NOTE: The adult mice (>8 weeks old) should be....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Extension tube	Top, Japan	X1-50	Connect with syringe and injection needle for antegrade perfusion.
Heater mat	Natsume Seisakusho, Japan	KN-475-3-40	Equipment to warm the perfusion plate.
Infusion pump	TERUMO, Japan	TE-311	Infusion syringe pump for antegrade perfusion.
Injection needle (27 gauge)	TERUMO, Japan	NN-2719S	Needle for insertion into the left ventricle.
Insulin (from bovine pancrease)	Sigma-Aldrich, USA	I5500	Dissolve in 0.1 M HCl.
Plastic syringe (20 mL)	TERUMO, Japan	SS-20ES	Use for infusion of CIB-EGTA.
Vascular clamp	Karl Hammacher GmbH, Germany	HSE 004-35	Small straight vascular clamp used for clamping aorta.
All other reagents	Nacalai Tesque, Japan