Extracellular Matrix-derived Foam Fabrication via Lyophilization: A Procedure to Generate Biological Scaffolds from ECM of Decellularized Tissues
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 method for synthesizing pure ECM-derived foams without the need for chemical crosslinking. These ECM-derived foams find applications as advanced 3D in vitro cell culture models or as pro-regenerative bioscaffolds.
Protocol
1. ECM-derived Foam Fabrication Prepare the homogenous ECM suspension by dispersing ECM fine powder in buffer solution and incubate at 37 °C with continuous agitation at 120 rpm until the suspension is warm. Dilute the ECM suspension in 0.2 M acetic acidic to the desired concentration. NOTE: Depending on the ECM source, stable foams may be prepared in the range of 10–100 mg/mL, with 15–50 mg/mL as the recommended range for DAT, DDT, and DLV. …
Offenlegungen
The authors have nothing to disclose.
Materials
| Analytical balance | Sartorius | CPA225D | |
| Centrifuge tubes (15 mL) | Sarstedt | 62.554.205 | |
| Centrifuge tubes (50 mL) | Sarstedt | 62.547.205 | |
| Collagen from bovine achilles tendon (insoluble) |
Sigma | C9879 | Or similar insoluble collagen source; Can be used as an alternative to decellularized tissues to fabricate the foams and microcarriers |
| Dessicator | Fisher Scientific | 8624426 | For lyophilized ECM and bioscaffold storage |
| Dewar flask | Fisher Scientific | 10-196-6 | Low form; volume range of 250 -500 mL |
| Double distilled water (ddH_{2} O) | From Barnstead GenPure xCAD Water Purification System |
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| D-PBS (Phosphate-buffered Saline) |
Wisent | 311425125 | |
| 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) |
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| Forceps | VWR | 37-501-32 | For transferring the foams |
| Freezer (-20 °C) | VWR | 97043-346 | |
| Freezer ( -80 °C) | Thermo Scientific | EXF40086A | |
| Glass vials | Fisher Scientific | 03-339-26D | To store lyophilized cryomilled ECM |
| Liquid nitrogen | For electrospraying | ||
| Lyophilizer | Labconco | 7750021 | FreeZone4.5 |
| 18 G needle | VWR | C ABD305185 | For dispensing ECM suspension into moulds |
| Orbital incubator shaker | SciLogex | 832010089999 | Temperature controlled (37 °C) |
| Serological pipettes (10 mL) | Sarstedt | 86.1254.001 | |
| Serological pipettes (25 mL) | Sarstedt | 86.1685.001 | |
| Sodium chloride | BioShop | 7647-14-5 | |
| Sodium phosphate monobasic | BioShop | 10049-21-5 |
Tags
Diesen Artikel zitieren
Extracellular Matrix-derived Foam Fabrication via Lyophilization: A Procedure to Generate Biological Scaffolds from ECM of Decellularized Tissues. J. Vis. Exp. (Pending Publication), e20485, doi: (2023).