Discovery of a Novel Polymer for Xeno-Free, Long-Term Culture of Human Pluripotent Stem Cell Expansion

Nasir, Aishah; Thorpe, Jordan; Burroughs, Laurence; Meurs, Joris; Pijuan?Galito, Sara; Irvine, Derek J.; Alexander, Morgan R.; Denning, Chris

Abstract

Human pluripotent stem cells (hPSCs) can be expanded and differentiated in vitro into almost any adult tissue cell type, and thus have great potential as a source for cell therapies with biomedical application. In this study, a fully-defined polymer synthetic substrate is identified for hPSC culture in completely defined, xenogenic (xeno)-free conditions. This system can overcome the cost, scalability, and reproducibility limitations of current hPSC culture strategies, and facilitate large-scale production. A high-throughput, multi-generational polymer microarray platform approach is used to test over 600 unique polymers and rapidly assess hPSC-polymer interactions in combination with the fully defined xeno-free medium, Essential 8 (E8). This study identifies a novel nanoscale phase separated blend of poly(tricyclodecane-dimethanol diacrylate) and poly(butyl acrylate) (2:1 v/v), which supports long-term expansion of hPSCs and can be readily coated onto standard cultureware. Analysis of cell-polymer interface interactions through mass spectrometry and integrin blocking studies provides novel mechanistic insight into the role of the E8 proteins in promoting integrin-mediated hPSC attachment and maintaining hPSC signaling, including ability to undergo multi-lineage differentiation. This study therefore identifies a novel substrate for long-term serial passaging of hPSCs in serum-free, commercial chemically-defined E8, which provides a promising and economic hPSC expansion platform for clinical-scale application.

Citation

Nasir, A., Thorpe, J., Burroughs, L., Meurs, J., Pijuan‐Galito, S., Irvine, D. J., Alexander, M. R., & Denning, C. (2020). Discovery of a Novel Polymer for Xeno-Free, Long-Term Culture of Human Pluripotent Stem Cell Expansion. Advanced Healthcare Materials, 10(6), Article 2001448. https://doi.org/10.1002/adhm.202001448

• Journal Article Type: Article
• Acceptance Date: Dec 9, 2020
• Online Publication Date: Dec 28, 2020
• Publication Date: Dec 28, 2020
• Deposit Date: Jan 7, 2021
• Publicly Available Date: Dec 29, 2021
• Journal: Advanced Healthcare Materials
• Print ISSN: 2192-2640
• Electronic ISSN: 2192-2659
• Publisher: Wiley
• Peer Reviewed: Peer Reviewed
• Volume: 10
• Issue: 6
• Article Number: 2001448
• DOI: https://doi.org/10.1002/adhm.202001448
• Keywords: Computer Networks and Communications; Pharmaceutical Science; Biomaterials; Biomedical Engineering
• Public URL: https://nottingham-repository.worktribe.com/output/5184113
• Publisher URL: https://onlinelibrary.wiley.com/doi/10.1002/adhm.202001448