Isolation and Culture of Murine Tongue Epithelial Cells: A Procedure to Isolate and Culture Tongue Epithelial Cells from Mouse Model
Published: April 30, 2023
Abstract
Source: Prasad, M. et al. In Vitro Establishment of a Genetically Engineered Murine Head and Neck Cancer Cell Line Using an Adeno-Associated Virus-Cas9 System. J. Vis. Exp. (2020)
In this video, we describe the isolation of epithelial cells from a murine tongue. These cells can be further cultured to give rise to cell colonies.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Isolation and Culture of Primary Cells Euthanize a 6-week-old male or female B6;129-Gt(ROSA)26Sortm1(CAG-cas9*,-EGFP)Fezh/J by CO₂ asphyxiation or any other IACUC approved protocol. NOTE: These CRISPR/Cas9 knockin mice have Cre recombinase-dependent expression of cas9 endonuclease …
Divulgaciones
The authors have nothing to disclose.
Materials
| Mouse strains | |||
| C57BL/6 | Envigo | ||
| B6;129-Gt(ROSA)26Sortm1(CAGcas9*,-EGFP)Fezh/J | Jackson labs | 24857 | |
| NOD.CB17-Prkdc-scid/NCr Hsd (Nod.Scid) | Envigo | ||
| Enzymes | |||
| Benzonase | Sigma | E1014 | |
| Collagenase IV | Thermo Fisher Scientific | 17104019 | |
| DNAse | Thermo Fisher Scientific | 18047019 | |
| Hyaluronidase | MilliporeSigma | H3506 | |
| Trypsin | Biological Industries Israel BeitHaemek Ltd. | 03-050-1B | |
| Culture Media, Chemicals and Reagents | |||
| BSA | Amresco | 0332-TAM-50G | |
| DMEM | Biological Industries Israel BeitHaemek Ltd. | 01-055-1A | |
| FBS | Biological Industries Israel BeitHaemek Ltd. | 04-127-1A | |
| HBSS | Sigma | H6648 | |
| PBS | Biological Industries Israel BeitHaemek Ltd. | 02-023-1A | |
| Tris buffer | MERCK Millipore | 648311-1KG | |
| Plastic wares | |||
| Amicon-ULTRA filter 100 KDa | Millipore | UFC910024 | |
| 0.22 µm sterile filters, 4 mm | Millex | SLGV004SL | |
| 0.45 µm sterile filters, 13 mm | Millex | SLHV013SL | |
| Culture plates | Greiner Bio-One | ||
| Falcon tubes | Greiner Bio-One |
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Isolation and Culture of Murine Tongue Epithelial Cells: A Procedure to Isolate and Culture Tongue Epithelial Cells from Mouse Model. J. Vis. Exp. (Pending Publication), e20471, doi: (2023).