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).