Tracheal decellularization using a combination of chemical, physical and bioreactor methods

Introduction: The purpose of this study was to compare different decellularization protocols with the conventional detergent enzymatic method (DEM) using continuous agitation. Methods: The first experiment compared conventional DEM with sonication and lyophilization+freeze-thaw cycles. A second expe...

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Bibliographic Details
Published in:International journal of artificial organs 2018-02, Vol.41 (2), p.100-107
Main Authors: Tchoukalova, Yourka D., Hintze, Justin M., Hayden, Richard E., Lott, David G.
Format: Article
Language:English
Subjects:
Bioreactors
Cartilage
Chondrocytes
Collagen
Deoxyribonucleic acid
DNA
Experiments
Extracellular matrix
Freeze drying
Freeze thaw cycles
Freeze-thawing
Glycosaminoglycans
Immunogenicity
Major histocompatibility complex
Organic chemistry
Quantitation
Sonication
Staining
Trachea
Weight
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Summary:Introduction: The purpose of this study was to compare different decellularization protocols with the conventional detergent enzymatic method (DEM) using continuous agitation. Methods: The first experiment compared conventional DEM with sonication and lyophilization+freeze-thaw cycles. A second experiment was carried out to compare time-adjusted DEM (2-hour instead of 4-hour incubations with 4% deoxycholate) to decellularization in a bioreactor. Cellularity was determined by DNA-quantitation, H&E-staining and immunostaining for major histocompatibility complex-1 (MHC-1). Results: Compared to untreated trachea, DNA content significantly decreased after 2 cycles in all groups in the first experiment and dropped below the minimal criteria for efficient decellularization (
ISSN:0391-3988
1724-6040
DOI:10.5301/ijao.5000648