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

Summary: 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....

Full description

Saved in:
Bibliographic Details
Main Authors: Khek-Chian Tham, Seong Soo Lim, Carine Bonnard, John E.A. Common
Format: Article
Language:English
Published: Elsevier 2025-03-01
Series:STAR Protocols
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166725000644
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667