Stabilizing milk-derived extracellular vesicles (mEVs) through lyophilization: a novel trehalose and tryptophan formulation for maintaining structure and Bioactivity during long-term storage
| dc.contributor.author | Dogan, Alan B. | en |
| dc.contributor.author | Marsh, Spencer R. | en |
| dc.contributor.author | Tschetter, Rachel J. | en |
| dc.contributor.author | Beard, Claire E. | en |
| dc.contributor.author | Amin, Md R. | en |
| dc.contributor.author | Jourdan, L. Jane | en |
| dc.contributor.author | Gourdie, Robert G. | en |
| dc.date.accessioned | 2025-01-22T13:45:47Z | en |
| dc.date.available | 2025-01-22T13:45:47Z | en |
| dc.date.issued | 2025-01-13 | en |
| dc.date.updated | 2025-01-19T04:12:46Z | en |
| dc.description.abstract | Extracellular vesicles (EVs) are widely investigated for their implications in cell-cell signaling, immune modulation, disease pathogenesis, cancer, regenerative medicine, and as a potential drug delivery vector. However, maintaining integrity and bioactivity of EVs between Good Manufacturing Practice separation/filtration and end-user application remains a consistent bottleneck towards commercialization. Milk-derived extracellular vesicles (mEVs), separated from bovine milk, could provide a relatively low-cost, scalable platform for large-scale mEV production; however, the reliance on cold supply chain for storage remains a logistical and financial burden for biologics that are unstable at room temperature. Herein, we aim to characterize and engineer a freeze-dried, mEV formulation that can be stored at room temperature without sacrificing structure/bioactivity and can be reconstituted before delivery. In addition to undertaking established mEV assays of structure and function on our preparations, we introduce a novel, efficient, high throughput assay of mEV bioactivity based on Electric Cell Substrate Impedance Sensing (ECIS) in Human dermal fibroblast monolayers. By adding appropriate excipients, such as trehalose and tryptophan, we describe a protective formulation that preserves mEV bioactivity during long-term, room temperature storage. Our identification of the efficacy of tryptophan as a novel additive to mEV lyophilization solutions could represent a significant advancement in stabilizing small extracellular vesicles outside of cold storage conditions. | en |
| dc.description.version | Published version | en |
| dc.format.mimetype | application/pdf | en |
| dc.identifier.citation | Journal of Biological Engineering. 2025 Jan 13;19(1):4 | en |
| dc.identifier.doi | https://doi.org/10.1186/s13036-024-00470-z | en |
| dc.identifier.uri | https://hdl.handle.net/10919/124299 | en |
| dc.language.iso | en | en |
| dc.rights | Creative Commons Attribution 4.0 International | en |
| dc.rights.holder | The Author(s) | en |
| dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | en |
| dc.title | Stabilizing milk-derived extracellular vesicles (mEVs) through lyophilization: a novel trehalose and tryptophan formulation for maintaining structure and Bioactivity during long-term storage | en |
| dc.title.serial | Journal of Biological Engineering | en |
| dc.type | Article - Refereed | en |
| dc.type.dcmitype | Text | en |