Protocol for reconstructing 3D human skin equivalents on endogenous dermal matrix with optional pigmentation

Area of Science:

• Biotechnology
• Tissue Engineering
• Dermatology

Background:

• Existing in vitro 3D skin models often utilize animal-derived collagen, which poorly mimics human dermal tissue.
• Current models face limitations in throughput, hindering large-scale research applications.

Purpose of the Study:

• To develop a robust protocol for reconstructing 3D human skin equivalents.
• To establish a method using endogenous dermal matrices, improving physiological relevance.
• To enable optional pigmentation for more comprehensive skin modeling.

Main Methods:

• Fibroblasts, keratinocytes, and melanocytes were expanded and seeded onto non-contractile polystyrene scaffolds.
• A novel air-lifting technique was employed for epidermal maturation.
• The protocol was optimized for 12- and 24-well formats for higher throughput.

Main Results:

• Successfully reconstructed 3D human skin equivalents with a fibroblast-derived dermal matrix.
• Achieved a fully stratified epidermis through epidermal maturation.
• Demonstrated the feasibility of incorporating optional pigmentation.

Conclusions:

• The developed protocol offers a more physiologically relevant alternative to animal collagen-based skin models.
• This method enhances throughput and allows for customizable skin equivalents, including pigmentation.
• The protocol provides a valuable tool for advancing skin research and development.