Fabrication of Extracellular Matrix from Mouse Model: An Effective Procedure to Prepare Decellularized Tongue Extracellular Matrix from Murine Tongue

Abstract

Source: Yao, Y. et al. Fabrication of Tongue Extracellular Matrix and Reconstitution of Tongue Squamous Cell Carcinoma In Vitro. J. Vis. Exp. (2018)

This video demonstrates the preparation of the murine tongue extracellular matrix by using the decellularization method, which involves exposing the harvested tongue to a series of physical, chemical, and enzymatic treatments. The resulting TEM scaffold retains its native extracellular matrix composition, removing the cellular components.

Protocol

All procedures involving animals have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Preparation of Tongue Extracellular Matrix (TEM)

1. Execute mice by cervical dislocation and remove the tongues using sterile surgical scissors and tweezers.
2. Immerse the tongues in 75% ethanol for 3 min, then put each tongue into a 1.5 mL Eppendorf (EP) tube with 1 mL of 10 mM sterile phosphate-buffered solution (PBS).
   NOTE: The concentration of PBS in all the following steps is the same as the concentration in this step.
3. Cell lysis....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
C57-BL/6J mice	Sun Yat-sen University Laboratory Animal Center
Ethanol	Guangzhou Chemical Reagent Factory	HB15-GR-2.5L
Sodium chloride	Sangon Biotech	A501218
Dibasic Sodium Phosphate	Guangzhou Chemical Reagent Factory	BE14-GR-500G
Potassium Phosphate Monobasic	Sangon Biotech	A501211
1.5 mL EP tube	Axygen	MCT-150-A
Ultra-low temperature freezer	Thermo Fisher Scientific
3.5 cm cell culture dish	Thermo Fisher Scientific	153066
6 cm cell culture dish	Greiner	628160
Triton X-100	Sigma-Aldrich	V900502
Calcium chloride	Sigma-Aldrich	746495
Magnesium chloride	Sigma-Aldrich	449164
DNase	Sigma-Aldrich	D5025
Ampicillin	Sigma-Aldrich	A9393
Surgical suture	Shanghai Jinhuan
250 mL wide-mouth bottle	SHUNIU	1407
Magnetic stirrer	AS ONE	1-4602-32
Carbon dioxide incubator	SHEL LAB	SCO5A
Water purifier	ELGA
Hepes free acid	BBI	A600264
4 °C fridge	Haier