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

Ahmed, Mahid; Hill, David; Ahmed, Shaheda; Przyborski, Stefan; Dalgarno, Kenneth; Dickinson, Anne

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.

Citation

Ahmed, M., Hill, D., Ahmed, S., Przyborski, S., Dalgarno, K., & Dickinson, A. (2024). Bioprinted autologous human skin equivalents for in vitro testing of therapeutic antibodies. International Journal of Bioprinting, 10(2), Article 1851. https://doi.org/10.36922/ijb.1851

• Journal Article Type: Article
• Acceptance Date: Jan 10, 2024
• Online Publication Date: Mar 11, 2024
• Publication Date: 2024
• Deposit Date: Jul 11, 2024
• Publicly Available Date: Jul 11, 2024
• Journal: International Journal of Bioprinting
• Print ISSN: 2424-7723
• Electronic ISSN: 2424-8002
• Publisher: AccScience
• Peer Reviewed: Peer Reviewed
• Volume: 10
• Issue: 2
• Article Number: 1851
• DOI: https://doi.org/10.36922/ijb.1851
• Public URL: https://durham-repository.worktribe.com/output/2525103