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…
Declarações
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)