A hybrid substratum for primary hepatocyte culture that enhances hepatic functionality with low serum dependency

Qingyuan Meng,^1–3 Chunsheng Tao,^1,4 Zhiye Qiu,¹ Toshihiro Akaike,³ Fuzhai Cui,¹ Xiumei Wang<sup>1

1</sup>School of Materials Science and Engineering, Tsinghua University, Beijing, People’s Republic of China; ²State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, People’s Republic of China; ³Biomaterials Center for Regenerative Medical Engineering, Ibaraki, Japan; ⁴The 401 Hospital of Chinese People’s Liberation Army, Qingdao, People’s Republic of China

Abstract: Cell culture systems have proven to be crucial for the in vitro maintenance of primary hepatocytes and the preservation of hepatic functional expression at a high level. A poly-(N-p-vinylbenzyl-4-O-β-D-galactopyranosyl-D-gluconamide) matrix can recognize cells and promote liver function in a spheroid structure because of a specific galactose–asialoglycoprotein receptor interaction. Meanwhile, a fusion protein, E-cadherin-Fc, when incubated with various cells, has shown an enhancing effect on cellular viability and metabolism. Therefore, a hybrid substratum was developed for biomedical applications by using both of these materials to combine their advantages for primary hepatocyte cultures. The isolated cells showed a monolayer aggregate morphology on the coimmobilized surface and displayed higher functional expression than cells on traditional matrices. Furthermore, the hybrid system, in which the highest levels of cell adhesion and hepatocellular metabolism were achieved with the addition of 1% fetal bovine serum, showed a lower serum dependency than the collagen/gelatin-coated surface. Accordingly, this substrate may attenuate the negative effects of serum and further contribute to establishing a defined culture system for primary hepatocytes.

Keywords: mouse primary hepatocytes, E-cadherin-Fc, PVLA, serum dependency, hybrid system