Cryomilling of Decellularized Tissue: A Technique to Obtain Fine Extracellular Matrix Powder

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)

In this video, we present the protocol to obtain finely ground extracellular matrix powder from lyophilized decellularized tissue by cryomilling. The powdered ECM retains its biochemical composition and can be used for microcarrier synthesis.

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 DAT, porcine DDT, and porcine DLV. Commercially available, insoluble collagen can also be used to fabricate foams and microcarriers, such as collagen sourced from bovine tendon, which has been used succ…

Açıklamalar

The authors have nothing to disclose.

Materials

Alligator clip leads VWR 470149-728
Aluminium foil Fisher Scientific 01-213-101
Analytical balance Sartorius CPA225D
Cryomilling system Retsch 20.745.0001 MM 400
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 (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
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
Milling chamber Retsch 02.462.0213 25 mL volume recommended
Milling balls VWR 16003-606 10 mm diameter, stainless steel recommended
Penicillin-streptomycin Life Technologies 15140-122 Used for proliferation media
Pipet-Aid XP Mandel Scientific DRU-4-000-101
Scoopula VWR 89259-968 For collecting microcarriers
Surgical scissors VWR 82027-590

Etiketler

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Bu Makaleden Alıntı Yapın
Cryomilling of Decellularized Tissue: A Technique to Obtain Fine Extracellular Matrix Powder. J. Vis. Exp. (Pending Publication), e20483, doi: (2023).

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