three-dimensional collagen-sponge-based culture system coated with simplified recombinant fibronectin improves the function of a hepatocyte cell line

• Published in: In vitro cellular & developmental biology. Animal Vol. 52; no. 3; pp. 271 - 277
• Main Authors: Nishida, Yuuki, Taniguchi, Akiyoshi
• Format: Journal Article
• Language: English
• Published: New York Springer US 01.03.2016; Springer Science & Business Media LLC; Society for In Vitro Biology
• Subjects: Albumins; Animal Genetics and Genomics; Animals; Biomedical and Life Sciences; BIOTECHNOLOGY; CCAAT-Enhancer-Binding Protein-alpha - biosynthesis; Cell Biology; Cell Culture; Cell culture techniques; Cell Culture Techniques - methods; Cell Line, Tumor; Cell lines; Collagen - metabolism; Collagens; Cultured cells; Cytochrome P-450 Enzyme System - metabolism; Developmental Biology; Fibronectins - genetics; Fibronectins - metabolism; Gene expression; Hep G2 Cells; Hepatocytes; Hepatocytes - metabolism; Honeycombs; Humans; Life Sciences; Messenger RNA; Porifera - metabolism; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Stem Cells; Tissue Scaffolds; 3D culture; Recombinant protein; Hepatocyte; Biomaterials
• ISSN: 1071-2690; 1543-706X
• DOI: 10.1007/s11626-015-9973-0

Hepatocytes are widely used in pharmaceutical drug discovery tests, but their hepatic functions decrease rapidly during in vitro culture. Many culture systems have been devised to address this problem. We here report that a three-dimensional (3D) collagen-based scaffold coated with simplified recombinant fibronectin (FN) enhanced the function of a hepatocyte cell line. The developed culture system uses a honeycomb collagen sponge coated with collagen-binding domain (CBD)-cell attachment site (CAS), a chimeric protein comprising the CBD and CAS of FN. The function of HepG2 cells grown on honeycomb collagen sponge coated with CBD-CAS was investigated by determining the messenger RNA (mRNA) expression levels of several genes. The mRNA expression level of albumin increased 3.25 times in cells grown on CBD-CAS-coated honeycomb collagen sponge for 3 days; the expression level of CCAAT/enhancer binding protein (C/EBPα) increased 40-fold after 1 d and up to 150-fold after 3 d. These results suggested that CBD-CAS-coated honeycomb collagen sponge could improve the functions of hepatocytes by inducing C/EBPα expression. The activation of cytochrome P450 (CYP) enzymes in HepG2 cells grown on CBD-CAS-coated honeycomb collagen sponge was measured at the mRNA level and was found to increase between two and six times compared to cells grown without the CBD-CAS coating, showing that this culture system induced CYP gene expression and thus may be useful in drug metabolism assays.