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 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 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) …
Divulgazioni
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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Citazione di questo articolo
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).