Isolation and Culture of Murine Tongue Epithelial Cells: A Procedure to Isolate and Culture Tongue Epithelial Cells from Mouse Model

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

1. 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 and EGFP directed by a CAG promoter. The upstream Lox-Stop-Lox (LSL) sequence present in the genome of these mice prevents the expression of Cas9 and ....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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