Isolation of Mouse Valve Interstitial Cells: An Enzymatic Method to Isolate and Culture VICs from Mouse Aortic Valve

Published: April 30, 2023

Abstract

Source: Bouchareb, R. et al. Isolation of Mouse Interstitial Valve Cells to Study the Calcification of the Aortic Valve In VitroJ. Vis. Exp. (2021)

In this video, we demonstrate the isolation of aortic valve interstitial cells from the heart of an adult mouse using a two-step collagenase procedure and their subsequent culture in low volume of growth media.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Preparation Before Valve Cell Isolation from Adult Mice Clean and sterilize all the surgical instruments shown in Figure 1A by using 70% v/v ethanol and subsequently autoclaving them for 30 min. Clean the surgical workspace with 70% ethanol. Add 500 µL of penicillin-streptomycin…

Representative Results

Figure 1: Description of valve dissection. (A) Representative image of all the surgical instruments needed for the dissection, scissors 2 is needed to open the skin of the mouse and scissors 3 to open the chest. Tweezers 5 and 6 are needed to hold the skin and open the chest. (B) …

Disclosures

The authors have nothing to disclose.

Materials

3 mm cutting edge scissors F.S.T 15000-00
Bonn Scissors F.S.T 14184-09
Collagenase type I (125 units/mg) Thermofisher Scientific 17018029
DMEM Thermofisher 11965092
Extra fine graefe forceps F.S.T 11150-10
FBS Gibco 16000044
Fine forceps F.S.T Dumont
HEPES 1 M solution STEMCELLS TECHNOLOGIES
L-Glutamine 100x Thermofisher Scientific A2916801
Mycozap Lanza VZA-2011 Mycoplasma elimination reagent
PBS 10x SIGMA-ALDRICH
Pencillin streptomycin 100x Thermofisher Scientific 10378016
Sodium Pyruvate 100 mM Thermofisher Scientific 11360070
Standard pattern forceps F.S.T 11000-12
Surgical Scissors – Sharp-Blunt F.S.T 14008-14
Trypsin 0.05% Thermofisher Scientific 25300054

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Cite This Article
Isolation of Mouse Valve Interstitial Cells: An Enzymatic Method to Isolate and Culture VICs from Mouse Aortic Valve. J. Vis. Exp. (Pending Publication), e20751, doi: (2023).

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