AccScience Publishing / IJB / Online First / DOI: 10.36922/ijb.1851
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RESEARCH ARTICLE

Bioprinted autologous human skin equivalents for in vitro testing of therapeutic antibodies

Mahid Ahmed1,2 David Hill3,4 Shaheda Ahmed2 Stefan Przyborski5,6 Kenneth Dalgarno1* Anne Dickinson2,4
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1 School of Engineering, Newcastle University, Newcastle upon Tyne, NE3 1PS, United Kingdom
2 Alcyomics Ltd., The Biosphere, Draymans Way, Newcastle Helix, Newcastle upon Tyne, NE4 5BX, United Kingdom
3 Faculty of Health Sciences and Wellbeing, Sunderland University, Sunderland, SR1 3SD, United Kingdom
4 Translational & Clinical Research Institute, Faculty of Medical Science, Medical School, Newcastle University, Newcastle upon Tyne, NE2 4HH, United Kingdom
5 Department of Bioscience, Durham University, South Road, Durham, DH1 3LE, United Kingdom
6 Reprocell Europe Ltd., NETPark Plexus, Thomas Wright Way, Sedgefield, Co. Durham, TS21 3FD, United Kingdom
IJB 2024, 10(2), 1851 https://doi.org/10.36922/ijb.1851
Submitted: 17 September 2023 | Accepted: 10 January 2024 | Published: 11 March 2024
© 2024 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
Abstract

In recent years, advances in tissue engineering have brought forward the accessibility of human skin equivalents for in vitro applications; however, the availability of human-based engineered tissue models suitable for high-throughput screening of biologics remains limited. Here, we report a method of manufacturing fully autologous (with both fibroblasts and keratinocytes from the same donor) human skin equivalents for determining preclinical therapeutic antibody adverse immune reactions in vitro. Using a combination of precise solenoid microvalve-based bioprinting and 96-well scale Alvetex inserts, autologous skin cells were bioprinted and cultured to develop a scalable approach to manufacturing skin equivalents. We demonstrated that fibroblasts and keratinocytes can be bioprinted with a high degree of precision while maintaining viability post printing. Histological staining showed that the bioprinted 96-well based skin equivalents were comparable to human skin. The fully autologous human skin equivalents were co-cultured in vitro with autologous peripheral blood monocytes with and without muromonab-CD3 (OKT3) and natalizumab (Tysabri), biologics which are known to cause and inhibit adverse immune reactions (type IV hypersensitivity), respectively. Analysis of supernatants from skin-equivalent monocyte co-cultures revealed significant proinflammatory cytokine responses (such as interferon gamma) in co-cultures treated with OKT3 when compared to Tysabri and negative controls. Consequently, this study provides proof of concept that through a combination of bioprinting and Alvetex scaffold-based culture systems, scalable human skin equivalents can be manufactured for high-throughput identification of adverse immune reactions during preclinical stages of the drug development process.

Keywords
Autologous tissue models
Skin-equivalent models
Microvalve bioprinting
Transwell culture
Funding
This research was supported by the EPSRC Centre for Doctoral Training in Additive Manufacturing and 3D Printing (EP/L01534X/1), and by Innovate UK through Project 103597 (Novel manufacture and commercialization of a 96-well 3D skin model for drug and toxicology testing).
Conflict of interest
The authors declare no conflicts of interest.
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International Journal of Bioprinting, Electronic ISSN: 2424-8002 Print ISSN: 2424-7723, Published by AccScience Publishing