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.
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.
The authors have nothing to disclose.
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 |