In Vitro Detection System to Evaluate the Immunogenic Potential of Xenografts

Tissue Engineering. Part C, Methods
Aila DaugsOliver Bloch

Abstract

Tissue antigenicity represents the main limitation for the use of xenografts in clinical practice. To eliminate xenoantigens and avoid graft rejection in human, decellularization is often used to remove all immunoreactive components from the extracellular matrix (ECM). After decellularization, acellular scaffolds are required to be investigated regarding the presence of antigens, but commonly used detection methods solely focus on known xenoantigens such as alpha Gal (Galα1,3-Galβ1-4GlcNAc-R) or major histocompatibility complex-I (MHC-I). However, there are unknown xenoantigens that escape the standard methods. To evaluate the immunological potential of xenogenic tissues, new in vitro methods need to be developed. Therefore, we established a novel human serum-based approach, including dot blot, Western blot, immunohistochemistry, and enzyme-linked immunosorbent assay (ELISA). With these methods, we analyzed protein extracts and tissue samples of native and decellularized bovine carotid arteries. All methods verified an effective removal of potential immunogens from the ECM through decellularization, and relative quantification with ELISA showed that 99.9% (p < 0.01) of antigenic components were successfully eliminated. We compa...Continue Reading

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Citations

Dec 24, 2018·Journal of the American Society of Nephrology : JASN·Zhiyong WangSteven C Borkan
Jul 7, 2019·Journal of Tissue Engineering and Regenerative Medicine·Chau Sang LauSwee-Hin Teoh
Dec 12, 2019·Journal of Tissue Engineering and Regenerative Medicine·Claudia BilodeauMartin Post

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Methods Mentioned

BETA
xenografts
chemical treatment
xenograft
electrophoresis
ELISA
dot blot
light microscopy
enzyme-linked immunosorbent assay
homograft

Software Mentioned

GraphPad Prism

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