Tham, K.-C., Lim, S. S., Bonnard, C., & Common, J. E. A. (2025). Protocol to reconstruct an in vitro 3D full-thickness skin equivalent model with collagen I in 6- and 12-well inserts. STAR Protocols, 6(1), 103658. https://doi.org/10.1016/j.xpro.2025.103658
Abstract:
In vitro 3D full-thickness reconstituted human skin has high physiological relevance due to the presence of differentiation features often lacking in 2D cell cultures. Here, we present a protocol to reconstruct a 3D skin model using human fibroblasts, keratinocytes, and rat-tail collagen I. We describe steps for cell expansion, the casting of cellular and acellular layers, seeding keratinocytes, the air lifting of culture, and incubation. We also demonstrate the use of chitosan to prevent tissue contraction in 12-well inserts. For complete details on the use and execution of this protocol, please refer to Robinson et al.
License type:
Attribution 4.0 International (CC BY 4.0)
Funding Info:
This research / project is supported by the A*STAR, National Research Foundation, and Economic Development Board - Industry Alignment Fund - Pre-Positioning - Skin Research Institute of Singapore
Grant Reference no. : H17/01/a0/004
This research / project is supported by the A*STAR, National Research Foundation, and Economic Development Board - Industry Alignment Fund - Pre-Positioning - Asian Skin Microbiome Program
Grant Reference no. : H18/01/a0/016
This research / project is supported by the A*STAR, National Research Foundation, and Economic Development Board - Industry Alignment Fund - Pre-Positioning - Asian Skin Microbiome Program 2.0
Grant Reference no. : H22/J1/a0/040
This research / project is supported by the A*STAR - Central Research Funding-Applied Translational Research (CRF-ATR)
Grant Reference no. :