Abstract
In vitro three-dimensional artificial human skin models (3D-HSMs) have revolutionized research in drug testing, disease modeling, and cosmetic evaluations. However, preserving the shelf-life of 3D-HSM during extended culture remains a challenge. Our previous study uncovered the anti-proliferative effect of fibroblasts exposed to low-molecular-weight fucoidan (LMW-F) at a lower concentration. Based on this study, we developed growth and differentiation media with LMW-F to extend 3D-HSM shelf-life. Two specifically designed media types supplemented with LMW-F components were formulated: (1) growth media (DFR-GM) consisting of DMEM (low glucose), Ham's F-12 K, and RPMI 1640 in a precise 2:2:1 ratio, with (LMW-F-DFR-GM) or without LMW-F (con-DFR-GM), and (2) differentiation media (DFR-DM) consisting of the same composition of DFR-GM with the addition of CaCl2, with (LMW-F-DFR-DM) or without LMW-F (con-DFR-DM). Human cell-based 3D-full-thickness HSE model (3D-FT-HSEM) was incubated with these media for 28 days. In comparison to the control group, the 3D-FT-HSEMs cultured in LMW-F supplemented medium showed a 47% reduction in diameter and retained 25% of their thickness (compared to a 94% reduction and complete degradation in the control group), along with enhanced expression of Ki67 proliferation marker (by approximately 20% at day 21) and reduced cell death. Overall, the present study provides valuable insights into enhancing the quality and extending the shelf-life of 3D-FT-HSEMs.