TABLE 2.

Common test methods for evaluation of the decellularization effect.

Evaluation index	Characterization modes	Test methods	Details
DNA	Quantitative	Commercial Kit	For DNA extraction and quantification
Nucleus	Localization	Immunofluorescence assessment and Histological staining	4′,6-diamidino-2-phenylindole (DAPI), Ematoxylin and Eosin
Collagen	Qualitative	Immunofluorescence assessment and Histological staining	Picro Sirius Red, Masson, Mallory and Van Gieson Trichromic staining and anti-collagen I, II, and III antibodies
	Quantitative	Experiments	Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS–PAGE), High-performance liquid chromatography with fluorescence detection (HPLC-FLD)
Elastin	Qualitative	Immunofluorescence assessment and Histological staining	Verhoeff and anti-elastin antibodies
	Quantitative	Experiments	NaOH extraction, Desmosine-base assay
Structural cell proteins	Qualitative	Immunofluorescence assessment	α-Smooth Muscle Actin (αSMA), β-Actin, Vimentin
Antigens	Qualitative	Immunohistochemistry assessment	Anti-Neu5Gc, anti α-Gal, and anti-SDa antibodies and Major Histocompatibility Complex (MHC)
	Quantitative	ELISA