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

Abstract

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.

Fabrication and Characterization of Heparin/Collagen Sponge for in Vitro Differentiation of Wharton’s Jelly-Derived Mesenchymal Stem Cells into Hepatocytes

“…This substance has many cell-binding motifs, low antigenicity, and high biocompatibility and biodegradability. While hepatocytes lose their functions (8) and proper morphology (9) after isolating and during the conventional culture procedures, collagen-based scaffolds can boost their function (10). Since collagen is one of the most important components of the liver ECM and given the fact that culturing the hepatocytes on the scaffolds containing liver ECM results in up-regulation of liver-specific genes (11), collagen-based materials have been shown to improve the differentiation of the stem cells toward hepatocytes (12).…”

Three-dimensional Culture Using Atelocollagen Sponge and Self-assembling Peptide Hydrogel

“…Atelocollagen sponge (ACS) was engineered to maintain structural strength. Some studies have focused on porous ACS 25,29,30) which was sterilized and cross-linked with formaldehyde and γ -rays (10 kGy) and lyophilized. Based on its physical strength, tissue affinity, affinity with SPH, and filling properties, it was decided that ACS would be a more suitable material for the present study than ceramic materials, which are not bio-absorptive and have substantial residual effects.…”

Heparin/Collagen 3D Scaffold Accelerates Hepatocyte Differentiation of Wharton’s Jelly-Derived Mesenchymal Stem Cells

“…Collagen type I, as a prominent part of Disse space [1], can be used to mimic the naïve hepatocyte niche. Collagen base scaffold has been demonstrated to improve the adult hepatocyte functions [9]. Well-developed bile canaliculi were found to form between primary hepatocytes cultured within collagen sandwich [10].…”