Development of high-throughput, reproducible, three-dimensional (3D) bioprinted skin equivalents (BPSEs) that are morphologically and functionally comparable to native skin tissue is advancing research in skin diseases, and providing a physiologically relevant platform for the development of therapeutics, transplants for regenerative medicine, and testing of skin products like cosmetics. Current protocols for the production of engineered skin grafts are limited in their ability to control 3D geometry of the structure and contraction leading to variability of skin function between constructs. In this study, we describe a method for the biofabrication of skin equivalents (SEs) that are fully bioprinted using an open-market bioprinter, made with commercially available primary cells and natural hydrogels. The unique hydrogel formulation allows for the production of a human-like SE with minimal lateral tissue contraction in a multiwell plate format, thus making them suitable for high-throughput bioprinting in a single print with fast print and relatively short incubation times. The morphology and barrier function of the fully 3D BPSEs are validated by immunohistochemistry staining, optical coherence tomography, and permeation assays. This article describes a method for the biofabrication of skin tissue equivalents in a multiwell plate format. The technique and results overcome shortcomings of previously published engineering methods, and show good architecture and barrier function from well to well; thus it may be used for compound functional testing and for the development of disease tissue models for screening.
- barrier function
- high-throughput screening