Disassembly of self-assembling peptide hydrogels as a versatile method for cell extraction and manipulation

Ligorio, Cosimo; Martinez-Espuga, Magda; Laurenza, Domenico; Hartley, Alex; Rodgers, Chloe B.; Kotowska, Anna M.; Scurr, David J.; Dalby, Matthew J.; Mata, Alvaro; Ordóñez-Morán, Paloma

Abstract

Self-assembling peptide hydrogels (SAPHs) are increasingly being used as two-dimensional (2D) cell culture substrates and three-dimensional (3D) matrices due to their tunable properties and biomimicry of native tissues. Despite these advantages, SAPHs often represent an end-point in cell culture, as isolating cells from them leads to low yields and disruption of cells, limiting their use and post-culture analyses. Here, we report on a protocol designed to easily and effectively disassemble peptide amphiphile (PA) SAPHs to retrieve 3D encapsulated cells with high viability and minimal disruption. Due to the pivotal role played by salt ions in SAPH gelation, tetrasodium ethylenediaminetetraacetic acid (Na4EDTA) was used as metal chelator to sequester ions participating in PA self-assembly and induce a rapid, efficient, clean, and gentle gel-to-sol transition. We characterise PA disassembly from the nano- to the macro-scale, provide mechanistic and practical insights into the PA disassembly mechanism, and assess the potential use of the process. As proof-of-concept, we isolated different cell types from cell-laden PA hydrogels and demonstrated the possibility to perform downstream biological analyses including cell re-plating, gene analysis, and flow cytometry with high reproducibility and no material interference. Our work offers new opportunities for the use of SAPHs in cell culture and the potential use of cells cultured on SAPHs, in applications such as cell expansion, analysis of in vitro models, cell therapies, and regenerative medicine.

Citation

Ligorio, C., Martinez-Espuga, M., Laurenza, D., Hartley, A., Rodgers, C. B., Kotowska, A. M., Scurr, D. J., Dalby, M. J., Ordóñez-Morán, P., & Mata, A. (2024). Disassembly of self-assembling peptide hydrogels as a versatile method for cell extraction and manipulation. Journal of Materials Chemistry B, 12(46), 11939-11952. https://doi.org/10.1039/D4TB01575D

• Journal Article Type: Article
• Acceptance Date: Oct 15, 2024
• Online Publication Date: Oct 17, 2024
• Publication Date: Dec 14, 2024
• Deposit Date: Oct 21, 2024
• Publicly Available Date: Oct 21, 2024
• Journal: Journal of Materials Chemistry B
• Print ISSN: 2050-750X
• Electronic ISSN: 2050-7518
• Publisher: Royal Society of Chemistry
• Peer Reviewed: Peer Reviewed
• Volume: 12
• Issue: 46
• Pages: 11939-11952
• DOI: https://doi.org/10.1039/D4TB01575D
• Public URL: https://nottingham-repository.worktribe.com/output/40850614
• Publisher URL: https://pubs.rsc.org/en/content/articlelanding/2024/tb/d4tb01575d