Hydrogel Microparticles Preserve Cell Secretome and Extracellular Vesicle Bioactivity for Improved Cell-Based Therapy Deployment

Hydrogel Microparticles Preserve Cell Secretome and Extracellular Vesicle Bioactivity for Improved Cell-Based Therapy Deployment | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Hydrogel Microparticles Preserve Cell Secretome and Extracellular Vesicle Bioactivity for Improved Cell-Based Therapy Deployment Li Ching Wong, Craig Jun Kit Wong, Kwan Yu Wu, Serene Ming En Chong, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9015333/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Secretomes derived from diverse progenitor cell classes are commonly employed in regenerative medicine applications to promote cell expansion and tissue differentiation. However, the bioactivity of secretome components, including soluble proteins and extracellular vesicles, is hindered by their low stability in basal cell culture media. Hydrogels are a promising strategy for improving secretome viability by providing a structurally supportive, biomimetic environment. This work examined the effects of poly(acrylic acid)-based hydrogel microparticles in the context of a previously characterised magnetic secretome-induction paradigm. We investigated the impact of a hydrogel microenvironment on the release and collection of secretome, during and after media conditioning. Various inorganic salt species were used to control microgel particle swelling and enable liquid phase collection. Calcium chloride was more effective than sodium or magnesium chlorides in facilitating secretome recovery with reference to promoting in vitro myogenesis at the cellular and protein levels. Notably, the secretome released from magnetically-induced myoblast cultures in microgel-derived conditioning media was as supportive of cell growth as foetal bovine serum (FBS), whereas the secretomes harvested from unstimulated cells in hydrogels or from stimulated cells in the absence of hydrogels were less effective. Microgel-derived conditioned media also enhanced the stability of isolated extracellular vesicles, better preserving their efficacy during storage at room temperature, 4° C and -20° C. These results indicate that these microgels protected the secretome from degradation. Media incorporating microgels represent an innovative approach with which to preserve the potency of cell secretomes collected following various induction protocols for improved commercial and clinical exploitation. conditioned media pulsed electromagnetic fields stem cells microgel regenerative medicine Full Text Additional Declarations The authors declare potential competing interests as follows: Author AF-O reports being a contributor to QuantumTx Pte. Ltd., which elaborates electromagnetic field devices for human use. Author AF-O is an inventor on patent WO 2019/17863 A1, System and Method for Applying Pulsed Electromagnetic Fields. Authors AF-O, CJKW, YKT, and RAJ are inventors on patent WO 2024/005714 A1, Directionally-specific Magnetic Modulation of the Cell Secretome for Medical and Commercial Applications. All Authors are inventors on patent WO/2025/212041, Collapsible cell-encapsulating scaffolds to enhance magnetically-induced secretome stabilisation and potency. There are no other known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Supplementary Files SupplementaryFileBlots.pdf All blots SupportingInformationnpj2026uploadready.docx Supplementary figures Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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