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 Mate…
Declarações
The authors have nothing to disclose.
Materials
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.