Bioscaffold Fabrication by Electrospraying: A Technique to Generate Microcarrier Scaffolds Derived from Extracellular Matrix of Decellularized Tissue

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.

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 the suspension and prevent clogging during electrospraying.
2. Load 3 mL of ECM suspension into a 3 mL Luer-Lock syringe and attach a winged infusion set onto the bore of the syringe.
   

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 .......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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