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 …

開示

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

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記事を引用
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).

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