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

Published: April 30, 2023

Abstract

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

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 Euthanize the mouse using a CO2 chamber. 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…

開示

The authors have nothing to disclose.

Materials

MICROSURGERY
6-0 suture, triangular section needle (Vicryl) Ethicon 6301124
9-0 micro-suture (Safil) B Braun G1048611
Adson forceps Fine Science Tools 11006-12
Adson forceps with teeth Fine Science Tools 11027-12
Castroviejo microneedle holder Fine Science Tools 12061-01
CO2 ventilation chamber for mouse euthanasia
Deionized water (Milli-Q IQ 7000, Ultrapure lab water system) Merck ZIQ7000T0
Disposable polystyrene tray (~30 × 50 cm)
Dissection microscope (Greenough, with two-armed gooseneck) Leica S6 D
Double-ended microspatula Fine Science Tools 10091-12
Dumont microforceps (two) Fine Science Tools 11252-20
Dumont microforceps with 45° tips Fine Science Tools 11251-35
Hair clippers Oster 76998-320-051
Halsey needle holder (with tungsten carbide jaws) Fine Science Tools 12500-12
Intravenous 24-gauge catheter (Insyte) BD 381512
Intravenous 26-gauge catheter (Terumo) Surflo-W SR+DM2619WX
Mayo scissors (tough cut, straight) Fine Science Tools 14110-15
Microforceps with ringed tips Aesculap FM571R
Micro-spring scissors (Vannas, curved) Fine Science Tools 15001-08
Needles (27 gauge; Microlance) BD 21018
Paper towel (sterile) or surgical napkin
Serrated scissors (CeramaCut, straight) Fine Science Tools 14958-09
Spatula (Freer-Yasargil) Aesculap OL166R
Syringes (1 mL; Plastipak) BD 3021001
Syringes (10 mL; Plastipak) BD 3021110
Tendon scissors (Walton) Fine Science Tools 14077-09
DECELLULARIZATION
70% Ethanol (absolute alcohol 99.9%); absolute alcohol must be adjusted to 70% (vol/vol) using deionized water Plum 1680766
Deionized water (Milli-Q IQ 7000, Ultrapure lab water system) Merck ZIQ7000T0
Luer-to-tubing male fittings (1/8 inch) World Precision Instruments 13158-100
Luer-to-tubing male fittings (1/8 inch) World Precision Instruments 13158-100
PBS (pH 7.4, 10×, Gibco) Thermo Fisher Scientific 70011044
Penicillin-streptomycin Gibco 15140122
Peristaltic pump (with 12 channels) Ole Dich 110AC(R)20G75
Silicone tubing (with 2-mm i.d. and 4 mm o.d.) Ole Dich 31399
Sodium Azide Sigma-Aldrich 08591-1ML-F
Sodium deoxycholate (DOC) Sigma-Aldrich D6750-100G

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記事を引用
Cardiopulmonary Complex Decellularization: A Technique for Decellularizing Heart and Lungs from Murine Model. J. Vis. Exp. (Pending Publication), e20739, doi: (2023).

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