Fabrication of Three-Dimensional HepG2 Liver Tissue by Centrifugation

Abstract

Background: We previously developed a rapid three-dimensional (3-D) tissue fabrication method using centrifugation. In this study, we fabricated 3-D HepG2 liver tissues using a scaffold-free centrifugation method and analyzed them biochemically.

Methods: We investigated the conditions for fabricating HepG2 tissues by varying the number of seeded cells (1.25-20 × 10⁶ cells/169 mm²). After two days of cultivation, cultured tissue cross-sections were examined. In addition, HepG2 tissues were cultured for an extended period (10 days), and biochemical metabolic analyses were performed.

Results: Thick HepG2 tissue was rapidly fabricated by centrifugation. Optical coherence tomography observations indicated that tissue thickness increased with a higher number of seeded cells. However, after two days of cultivation, biochemical analyses and observations of the paraffin section indicated that tissues fabricated with over 5.0 × 10⁶ cells were weak and damaged. However, the liver tissues fabricated using 1.25-2.5 × 10⁶ cells were compact with high cell density. HepG2 tissue was cultured for 10 days; however, albumin production remained minimal.

Conclusion: This study established optimal conditions for the rapid fabrication of HepG2 liver tissue. The proposed method holds significant potential for advancing liver tissue engineering and hepatocyte biology.