ECM Suspension Preparation: A Technique to Obtain Homogenous Suspension from Tissue-derived Extracellular Matrix

Abstract

Source: Kornmuller, A. et al. Fabrication of Extracellular Matrix-derived Foams and Microcarriers as Tissue-specific Cell Culture and Delivery Platforms. J. Vis. Exp. (2017)

This video describes the preparation of a homogenized and digested extracellular matrix suspension from decellularized and lyophilized tissues. This technique helps in fabricating ECM-derived porous foams and microcarriers for use as substrates in advanced 3D cell culture models.

Protocol

1. Decellularized Tissue Processing Decellularization and lyophilization Decellularize tissue(s) of interest following an established protocol. NOTE: Scaffolds in the current study have been prepared following published decellularization protocols for human decellularized adipose tissue (DAT), porcine decellularized dermal tissue (DDT), and porcine decellularized left ventricle (DLV). Commercially available, insoluble collagen can also be used to fabricate…

Materials

Acetic acid, glacial	BioShop	ACE222.5
α-amylase	Sigma	101074694	from Aspergillus aryzae
Centrifuge 75004251	Thermo Scientific		With swinging bucket rotor for 15 & 50 mL centrifuge tubes
Centrifuge tubes (15 mL)	Sarstedt	62.554.205
Dessicator	Fisher Scientific	8624426	For lyophilized ECM and bioscaffold storage
Double distilled water (ddH2O)			From Barnstead GenPure xCAD Water Purification System
ECM (Extracellular Matrix)			Isolated from human adipose tissue, porcine dermis or porcine myocardium, as described in Flynn et al. 2010, Reing et al. 2010, and Wainwright et al. 2010 (ref # 28, 8, 32)
Hand held homogenizer	Fisher Scientific	14-359-251	Speed: 8,000 – 30,000 rpm
Homogenizer accessories: saw tooth bottom generator probes	Fisher Scientific	14-261-17	10 x 95 mm
Liquid nitrogen
Sodium chloride	BioShop	7647-14-5
Sodium phosphate monobasic	BioShop	10049-21-5
Surgical scissors	VWR	82027-590