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

Abstract

Source: Bouchareb, R. et al. Isolation of Mouse Interstitial Valve Cells to Study the Calcification of the Aortic Valve In Vitro. J. 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

1. 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.
2. Add 500 µL of penicillin-streptomycin to 50 mL of 10 mm HEPES. Prepare an aliquot of 50 mL of 1x phosphate-buffered saline (PBS). Keep the solutions on ice.
3. Prepare 1 mg/mL and 4.5 mg/mL ....

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 scissor.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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