Bioscaffold Fabrication by Electrospraying: A Technique to Generate Microcarrier Scaffolds Derived from Extracellular Matrix of Decellularized Tissue
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 demonstrates the procedure of extracellular matrix-derived microcarrier fabrication by electrospraying. These microcarriers simulate the native tissue-specific microenvironment and are stable in long-term 3D in vitro cell culture models.
Protocol
1. ECM-derived Microcarrier Fabrication via Electrospraying NOTE: An overview of the electrospraying set-up is shown in Figure 1. Incubate a previously prepared cryomilled ECM suspension at 37 °C with continuous agitation at 100 rpm O/N. NOTE: Cryomilled ECM is used to fabricate the ECM-derived microcarriers to ensure greater uniformity in t…
Representative Results
Figure 1. Overview of the electrospraying apparatus used in microcarrier fabrication. A: Image showing the arrangement of the key electrospraying equipment including the syringe pump and high voltage power supply, as well as the positioning of the needle relative to the Dewar of liquid nitrogen. B: Electrospraying schematic, including the recommended ranges …
Divulgations
The authors have nothing to disclose.
Materials
| Alligator clip leads | VWR | 470149-728 | |
| Aluminium foil | Fisher Scientific | 01-213-101 | |
| Centrifuge tubes (15 mL) | Sarstedt | 62.554.205 | |
| Centrifuge tubes (50 mL) | Sarstedt | 62.547.205 | |
| 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 |
| 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) | ||
| Liquid nitrogen | For electrospraying | ||
| Lyophilizer | Labconco | 7750021 | FreeZone4.5 |
| Orbital incubator shaker | SciLogex | 8.3201E+11 | Temperature controlled (37 °C) |
| Retort stand | VWR | 470019-526 | |
| Retort stand clamp | VWR | 21573-606 | |
| Safety-Lok Syringe | BD | 309606 | 3 mL Luer lock syringe for microcarrier fabrication and dispensing ECM suspension |
| Scoopula | VWR | 89259-968 | For collecting microcarriers |
| Syringe pump | VWR | 10117-490 | Microprocessor controlled |
| High voltage power supply | Gamma High Voltage Research | ES30P-5W/DDPM | Capable of recommended 15 – 25 kV voltage range |
| Winged infusion set | Terumo | 22258092 | 30 cm tubing length, 25 G 3/4" recommended; Other needle gauges can be used and may influence microcarrier diameter |
Tags
Citer Cet Article
Bioscaffold Fabrication by Electrospraying: A Technique to Generate Microcarrier Scaffolds Derived from Extracellular Matrix of Decellularized Tissue. J. Vis. Exp. (Pending Publication), e20486, doi: (2023).