Abstract: Objective This study aimed to fabricate and to characterize decellularized human umbilical cord-derived scaffolds (dHUCS), to evaluate the function of rodent hepatocyte-derived liver progenitor-like cells (rHepLPCs) cultured on 3-dimensional (3D) scaffolds, and to provide a novel liver tissue engineering protocol. Methods Hepatocytes were isolated from mice by a 2-step collagenase perfusion method, expanded in vitro and labeled with Dil. The dHUCS was characterized by detection of component, Masson staining and hematoxylin and eosin staining. The function of 3D-cultured rHepLPCs was analyzed by quantitative polymerase chain reaction and biochemical assays. Results After decellularization, the nucleic acid residue of umbilical cord-derived scaffolds was 60.20 ± 1.42 ng/mg (dry weight), and the removal rate of nucleic acids was 90.5%. Matrix component were rich. After 2-week culture, albumin, glucose-6-phosphatase and carbamoyl-phosphate synthetase 1 expression levels of hepatocytes were upregulated by 10.1 ± 2.3, 34.4 ± 1.3 and 42.0 ± 22.0 times, respectively. As for ammonia metabolism, the ammonia clearance and urea synthesis levels were 5.7 ± 0.9 mg/(dL·million cells·day) and 7.1 ± 2.5 mg/(dL·million cells·day) in 2-dimensional-cultured rHepLPCs, 26.7 ± 4.5 mg/(dL·million cells·day) and 42.1 ± 7.0 mg/(dL·million cells·day) in 3D-cultred rHepLPCs, respectively. Conclusion The dHUCS is an ideal liver tissue engineering material. The 3D cultured rHepLPCs could have better function, which have potential therapeutic effect on acute liver failure characterized by elevated serum ammonia.

Key words: Decellularized human umbilical cord-derived scaffold, Decellularization, Rodent hepatocyte-derived liver progenitor-like cell, Transformation and expansion medium, 3-dimensional