Decellularized Porcine Esophageal Scaffold Preparation: A Technique to Generate Acellular Scaffold from a Pig Esophagus

Abstract

Source: Green, N. H. et al. Production, characterization and potential uses of a 3D tissue-engineered human esophageal mucosal model. J. Vis. Exp. (2015)

In this video, we demonstrate the preparation of a decellularized pig esophagus scaffold, with its biochemical architecture and composition preserved, for tissue engineering applications.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Preparation of Decellularized Esophageal Scaffold Obtain intact porcine esophagi at an abattoir from freshly slaughtered Landrace pigs that are used for food production. One single esophagus will provide sufficient scaffolds for approximately 30 separate constructs. NOTE: Due to the extensive and…

Materials

DMEM	Labtech	LM-D1112	Warm in 37 °C water bath before use
Ham's F12	Labtech	LM-H1236	Warm in 37 °C water bath before use
Foetal Calf Serum	Labtech	FB-1090
Epidermal Growth Factor	R+D Systems	236-EG-200	Prepare 200 µg/ml stock solution in 10 mM acetic acid, 1% FCS
Cholera toxin	Sigma-Aldrich	C8052	Prepare stock solution in water
L-Glutamine	Sigma-Aldrich	G7513
Penicillin-Streptomycin	Sigma-Aldrich	P0781
Amphotericin B	Gibco	15290-026	Brand name Fungizone
PBS	Oxoid	BR0014	Dissolve 1 tablet in 100 ml water and autoclave to sterilize
Collagenase A	Roche	10103578001
Povidone-iodine solution	Ecolab	10830E	Brand name Videne
Ethanol	Sigma-Aldrich	E7023
Chelex 100	Sigma-Aldrich	C7901
Newborn calf serum	Gibco	26010074
15 ml universal tube	SLS	SLS7504
180 ml pot	VWR	216-2603