Scalable Production and Isolation of Extracellular Vesicles: Available Sources and Lessons from Current Industrial Bioprocesses

Abstract

Potential applications of extracellular vesicles (EVs) are attracting increasing interest in the fields of medicine, cosmetics, and nutrition. However, the manufacturing of EVs is currently characterized by low yields. This limitation severely hampers progress in research at the laboratory and clinical scales, as well as the realization of successful and cost-effective EV-based products. Moreover, the high level of heterogeneity of EVs further complicates reproducible manufacturing on a large scale. In this review, possible directions toward the scalable production of EVs are discussed. In particular, two strategies are considered: i) the optimization of upstream unit operations and ii) the exploitation of well-established and mature technologies already in use in other industrial bioprocesses.

Nanoalgosomes: Introducing extracellular vesicles produced by microalgae

“…Three independent biological replicas were performed F I G U R E  The scalability, sustainability and economical viability of nanoalgosome production compared to the most common sources of therapeutic vesicles. [The ranking represents an arbitrary evaluation based on the analysis of the nanoalgosome production in the context of current bioprocesses (Paganini et al, 2019)] bioremediation developments and biosynthesis of high-added value biochemicals (Blanch, 2012;Khozin-Goldberg et al, 2016;Pulz & Gross, 2004;Sun et al, 2013;Yáñez-Mó et al, 2015). Recent advances in microalgal biotechnology have generated much optimism for a viable industrial production of microalgae-derived compounds such as antioxidants or omega-3 polyunsaturated fatty acids.…”

<p>Extracellular Vesicles As Nanomedicine: Hopes And Hurdles In Clinical Translation</p>

“…Isolation methods and equipment processes considered for commercial-scale downstream isolation of EVs from conditioned medium or cell suspensions are most likely going to conservatively follow, at least for the time being, methodologies already well-established in the biotech industry for producing other therapeutic biological products. 24 The production sequence of EVs typically combines low-speed centrifugation to pelletize and remove cells/cell debris suspended in the starting raw material, followed by sequential centrifugation or filtration for clarification. 25 While earlier developed laboratory-scale processes typically used ultracentrifugation at 20,000~100,000 ×g for several hours to pelletize EVs, now concentration by tangential flow-filtration (TFF) followed by size-exclusion chromatography (SEC) is becoming the preferred approach to better preserve the functional activity of EVs.…”

Mesenchymal Stem Cell-Derived Extracellular Vesicles: Opportunities and Challenges for Clinical Translation

“…However, sequential centrifugation steps have been used for large-scale production of clinical-grade MSC-EVs ( Mendt et al, 2018 ). Size-based fractionation methods that include tangential flow filtration and size exclusion chromatography are GMP-compliant and scalable systems for EV isolation [for reviews, see Agrahari et al (2019) ; Paganini et al (2019) ]. Ultrafiltration also reduces the isolation times and costs compared with other techniques ( Saxena et al, 2009 ; Bari et al, 2018 , 2019b , c ).…”