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

Published: April 30, 2023

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…

Divulgaciones

The authors have nothing to disclose.

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

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ECM Suspension Preparation: A Technique to Obtain Homogenous Suspension from Tissue-derived Extracellular Matrix. J. Vis. Exp. (Pending Publication), e20484, doi: (2023).

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