Abstract
Improvement of three-dimensional (3D) culturing conditions, including the substrates used for cell growth, is needed for a variety of cell-based applications as alternatives to animal testing and experimentation. In this study, we synthesized a nonwoven fabric comprised of polyacrylonitrile nanofibers of several widths for use as scaffolds in 3D culturing, and evaluated the optimal nanofiber width to achieve growth and function of human hepatoblasts derived from human fetal hepatocytes during static culturing. For these analyses, several nanofibers with widths between 170–1300 nm were generated by electrospinning. When cultured within 600 nm nanofiber scaffolds, hepatoblasts derived from human fetal hepatocytes exhibited a spherical shape. Furthermore, these cells exhibited enhanced cellular CYP3A4 activity when cultured in 300 nm and 600 nm nanofibers, reaching 2.20 ± 0.05 pmol/106 cells/min, and 2.63 ± 0.06 pmol/106 cells/min, respectively, which were comparable to those of primary human hepatocytes harvested from livers of adult donors. Conversely, hepatoblasts exhibited a flat shape and relatively low CYP3A4 activity when cultured within 170 nm, 1000 nm, and 1300 nm nanofibers. These results suggest that there is a link between the shape and activity of hepatocytes, and that our nonwoven fabric made with polyacrylonitrile nanofibers comprises a promising scaffold for 3D culturing of human hepatocytes.
