Method Article

Cardiopulmonary Complex Decellularization: A Technique for Decellularizing Heart and Lungs from Murine Model

July 8th, 2025

In This Article

Abstract

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Source: Mayorca-Guiliani, A. E. et al. Decellularization of the Murine Cardiopulmonary Complex. J. Vis. Exp. (2021)

This video demonstrates a method of producing decellularized extracellular matrix or ECM scaffolds by applying antegrade perfusion of detergents through the vasculature. The structural ECM scaffolds can then be used to analyze ECM remodeling associated with cardiopulmonary disease.

Protocol

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

1. Post-mortem Microsurgery

  1. Euthanize the mouse using a CO2 chamber.
    1. Place mouse into a 4 L chamber, and begin to fill with 100% CO2, starting at 0.2 L/min for 2 min, increasing until reaching a flow of 0.8 L/min after 3 min. The mouse should fall unconscious during the first 2 min, and then respiration should cease (usually around 5 min, but flow can be maintained as necessary). Confirm death prior to proceedin....

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Disclosures

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No conflicts of interest declared.

Materials

List of materials used in this article
NameCompanyCatalog NumberComments
MICROSURGERY
6-0 suture, triangular section needle (Vicryl)Ethicon6301124
9-0 micro-suture (Safil)B BraunG1048611
Adson forcepsFine Science Tools11006-12
Adson forceps with teethFine Science Tools11027-12
Castroviejo microneedle holderFine Science Tools12061-01
CO2 ventilation chamber for mouse euthanasia
Deionized water (Milli-Q IQ 7000, Ultrapure lab water system)MerckZIQ7000T0
Disposable polystyrene tray (~30 × 50 cm)
Dissection microscope (Greenough, with two-armed gooseneck)LeicaS6 D
Double-ended microspatulaFine Science Tools10091-12
Dumont microforceps (two)Fine Science Tools11252-20
Dumont microforceps with 45° tipsFine Science Tools11251-35
Hair clippersOster76998-320-051
Halsey needle holder (with tungsten carbide jaws)Fine Science Tools12500-12
Intravenous 24-gauge catheter (Insyte)BD381512
Intravenous 26-gauge catheter (Terumo)Surflo-WSR+DM2619WX
Mayo scissors (tough cut, straight)Fine Science Tools14110-15
Microforceps with ringed tipsAesculapFM571R
Micro-spring scissors (Vannas, curved)Fine Science Tools15001-08
Needles (27 gauge; Microlance)BD21018
Paper towel (sterile) or surgical napkin
Serrated scissors (CeramaCut, straight)Fine Science Tools14958-09
Spatula (Freer-Yasargil)AesculapOL166R
Syringes (1 mL; Plastipak)BD3021001
Syringes (10 mL; Plastipak)BD3021110
Tendon scissors (Walton)Fine Science Tools14077-09
DECELLULARIZATION
70% Ethanol (absolute alcohol 99.9%); absolute alcohol must be adjusted to 70% (vol/vol) using deionized waterPlum1680766
Deionized water (Milli-Q IQ 7000, Ultrapure lab water system)MerckZIQ7000T0
Luer-to-tubing male fittings (1/8 inch)World Precision Instruments13158-100
Luer-to-tubing male fittings (1/8 inch)World Precision Instruments13158-100
PBS (pH 7.4, 10×, Gibco)Thermo Fisher Scientific70011044
Penicillin-streptomycinGibco15140122
Peristaltic pump (with 12 channels)Ole Dich110AC(R)20G75
Silicone tubing (with 2-mm i.d. and 4 mm o.d.)Ole Dich31399
Sodium AzideSigma-Aldrich08591-1ML-F
Sodium deoxycholate (DOC)Sigma-AldrichD6750-100G

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Tags

Cardiopulmonary ComplexDecellularization TechniqueMurine ModelAntegrade PerfusionDetergent SolutionsDeionized WaterDOC PerfusionSDS PerfusionTracheal CatheterizationAortic Arch Catheter

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