Related Experiment Video
Updated: Aug 6, 2026

Building Up Skin Models for Numerous Applications - from Two-Dimensional (2D) Monoculture to Three-Dimensional (3D) Multiculture
Published on: October 20, 2023
Protocol for reconstructing 3D human skin equivalents on endogenous dermal matrix with optional pigmentation
Khek-Chian Tham1, Zhi Heng Lau1, Joycelyn X Y Lee1
1A∗STAR Skin Research Labs (A∗ SRL), Agency for Science, Technology and Research (A∗STAR), 11 Mandalay Road, #17-01 Clinical Sciences Building, Singapore 308232, Singapore.
Abstract:
Previous in vitro 3D skin models are limited by their reliance on animal-derived collagen, a poor representation of the human dermis, and low throughput. Here, we present a protocol for reconstructing 3D human skin equivalents on an endogenous dermal matrix, in 12- and 24-well format, with optional pigmentation. We describe the expansion and seeding of fibroblasts, keratinocytes, and melanocytes in non-contractile polystyrene scaffolds. We then detail air-lifting and epidermal maturation, resulting in a fibroblast-derived dermal matrix with a fully stratified epidermis. For complete details on the execution of this protocol, please refer to Tham et al.1.

