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Protocol to reconstruct an in vitro 3D full-thickness skin equivalent model with collagen I in 6- and 12-well inserts

Lookup NU author(s): Professor John Common

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Abstract

© 2025 The Author(s). 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.1,2

Publication metadata

Author(s): Tham K-C, Lim SS, Bonnard C, Common JEA

Publication type: Article

Publication status: Published

Journal: STAR Protocols

Year: 2025

Volume: 6

Issue: 1

Print publication date: 21/03/2025

Online publication date: 04/03/2025

Acceptance date: 02/04/2018

Date deposited: 23/01/2026

ISSN (electronic): 2666-1667

Publisher: Cell Press

URL: https://doi.org/10.1016/j.xpro.2025.103658

DOI: 10.1016/j.xpro.2025.103658

PubMed id: 40042969

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