Cells for EV production: Evercyte produces extracellular vesicles from any cell type of interest; please select from our broad product catalogue of telomized human cells or send us the cell line(s) you are interested in. Production systems:
Evercyte produces extracellular vesicles from 2D standard cell culture systems. Alternatively, we use hollow fiber bioreactor systems for continuous production of extracellular vesicles over a peroid of at least 10 weeks. Small cartridges with 20 kD MWCO polysulfone fibers with a surface area of 450 cm² are used to set up the system. Thereafter, medium cartridges with 4000 cm² are implemented for continuous EV production (FiberCell Systems
). During the production phase glucose, glutamine consumption and secretion of metabolites such as ammonium are monitored.
EV enrichment and purification:
cell culture supernatants are processed using ultra centrifugation (UZ), tangential flow filtration (TFF) and/or size exclusion chromatography (SEC).
EVs production and stability of enriched EV preparations
hollow fiber bioreactor
Extracellular vesicles from mesenchymal stem cells are produced in a hollow fiber bioreactor system with continuous harvests over a peroid of at least 4 months. After enrichment from supernatants using TFF, the EVs, harvested at different time points during the production phase (1, 3, 9, 11 weeks), have been characterized for their miRNA cargo. As shown in the right graph, EVs show by a similar miRNA cargo (selection of miRNAs is shown) demonstrating that the hollow fiber bioreactor system allows stable EV production.
Metabolic control of EV production process
monitoring glucose / glutamine consumtion and accumulation of metabolites
In order to ensure an optimal supply of the cells with nutrients during the production process, the consumption of glucose and glutamine is continuously monitored. An optimal feeding strategy is developed accordingly. The content of certain metabolites such as ammonium is also measured regularly.
Leaflets - extracellular vesicles
Gobin J, Muradia G, Mehic J, Westwood C, Couvrette L, Stalker A, Bigelow S, Luebbert CC, Bissonnette FS, Johnston MJW, Sauvé S, Tam RY, Wang L, Rosu-Myles M, Lavoie JR. (2021) Hollow-fiber bioreactor production of extracellular vesicles from human bone marrow mesenchymal stromal cells yields nanovesicles that mirrors the immuno-modulatory antigenic signature of the producer cell. Stem Cell Res Ther. 2021 Feb 12;12(1):127
<span ">Cao J, Wang B, Tang T, Lv L, Ding Z, Li Z, Hu R, Wei Q, Shen A, Fu Y, Liu B. (2020) Three-dimensional culture of MSCs produces exosomes with improved yield and enhanced therapeutic efficacy for cisplatin-induced acute kidney injury. Stem Cell Res Ther. 2020 May 27;11(1):206. https://pubmed.ncbi.nlm.nih.gov/32460853/
Watson DC, Yung BC, Bergamaschi C, Chowdhury B, Bear J, Stellas D, Morales-Kastresana A, Jones JC, Felber BK, Chen X, Pavlakis GN. Scalable, cGMP-compatible purification of extracellular vesicles carrying bioactive human heterodimeric IL-15/lactadherin complexes. J Extracell Vesicles. 2018 Feb 28;7(1):1442088