Large‐scale production of extracellular vesicles: Report on the “massivEVs” ISEV workshop

Paolini, Lucia; Monguió‐Tortajada, Marta; Costa, Marta; Antenucci, Fabio; Barilani, Mario; Clos‐Sansalvador, Marta; Andrade, André Cronemberger; Giancaterino, Sara; Kronstadt, Stephanie M.; Mizenko, Rachel R.; Nawaz, Muhammad; Osteikoetxea, Xabier; Pereira, Carla; Shrivastava, Surya; Boysen, Anders T.; Wakker, Simonides Immanuel van de; Herwijnen, Martijn J. C. van; Wang, Xiaoqin; Watson, Dionysios C.; Gimona, Mario; Kaparakis‐Liaskos, Maria; Konstantinov, Konstantin N.; Lim, Sai Kiang; Meisner‐Kober, Nicole; Stork, Michiel; Nejsum, Peter; Radeghieri, Annalisa; Rohde, Eva; Touzet, Nicolas; Wauben, Marca H. M.; Witwer, Kenneth W.; Bongiovanni, Antonella; Bergese, Paolo

doi:10.1002/jex2.63

Cited by 36 publications

(35 citation statements)

References 55 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This not only includes lacking standardization of culture conditions during sEV production but also of sEV isolation methods. In addition, changing from flaskbased expansion processes to a (semi-) automated expansion in bioreactors is still hampered and thus prevents efficient upscaling strategies (Paolini et al, 2022).…”

Section: Discussionmentioning

confidence: 99%

“…Translation of this yet laboratory-scale optimized process toward a GMP-compliant manufacturing for clinical applications could be supported by quality by design (QbD) approaches. Critical process parameters and steps could be identified and different quality controls (e.g., microbial or endotoxin testing) should be included in the whole manufacturing process (Yu et al, 2014;Paolini et al, 2022).…”

Section: Figurementioning

confidence: 99%

See 1 more Smart Citation

A novel approach for large-scale manufacturing of small extracellular vesicles from bone marrow-derived mesenchymal stromal cells using a hollow fiber bioreactor

Jakl

Ehmele

Winkelmann

et al. 2023

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Mesenchymal stromal cells (MSCs) are promising therapeutic candidates in a variety of diseases due to having immunomodulatory and pro-regenerative properties. In recent years, MSC-derived small extracellular vesicles (sEVs) have attracted increasing interest as a possible alternative to conventional cell therapy. However, translational processes of sEVs for clinical applications are still impeded by inconsistencies regarding isolation procedures and culture conditions. We systematically compared different methods for sEV isolation from conditioned media of ex vivo expanded bone marrow-derived MSCs and demonstrated considerable variability of quantity, purity, and characteristics of sEV preparations obtained by these methods. The combination of cross flow filtration with ultracentrifugation for sEV isolation resulted in sEVs with similar properties as compared to isolation by differential centrifugation combined with ultracentrifugation, the latter is still considered as gold standard for sEV isolation. In contrast, sEV isolation by a combination of precipitation with polyethylene glycol and ultracentrifugation as well as cross flow filtration and size exclusion chromatography resulted in sEVs with different characteristics, as shown by surface antigen expression patterns. The MSC culture requires a growth-promoting supplement, such as platelet lysate, which contains sEVs itself. We demonstrated that MSC culture with EV-depleted platelet lysate does not alter MSC characteristics, and conditioned media of such MSC cultures provide sEV preparations enriched for MSC-derived sEVs. The results from the systematic stepwise evaluation of various aspects were combined with culture of MSCs in a hollow fiber bioreactor. This resulted in a strategy using cross flow filtration with subsequent ultracentrifugation for sEV isolation. In conclusion, this workflow provides a semi-automated, efficient, large-scale-applicable, and good manufacturing practice (GMP)-grade approach for the generation of sEVs for clinical use. The use of EV-depleted platelet lysate is an option to further increase the purity of MSC-derived sEVs.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Figurementioning

confidence: 99%

A novel approach for large-scale manufacturing of small extracellular vesicles from bone marrow-derived mesenchymal stromal cells using a hollow fiber bioreactor

Jakl

Ehmele

Winkelmann

et al. 2023

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

show abstract

“…However, each step of scalable EV manufacturing under GMP compliance is still exposed to several challenges ( 163 , 164 ). These challenges exist not only in upstream and downstream stages ( 165 , 166 ) of EV production and isolation, such as cell culture and purification steps, respectively, but also during EV storage ( 108 ), quality control ( 167 ), and functional evaluation steps ( 148 , 168 ).…”

Section: Extracellular Vesicles a New Therapeutic Paradigm For Autoim...mentioning

confidence: 99%

Extracellular vesicles and their cells of origin: Open issues in autoimmune diseases

et al. 2023

View full text Add to dashboard Cite

The conventional therapeutic approaches to treat autoimmune diseases through suppressing the immune system, such as steroidal and non-steroidal anti-inflammatory drugs, are not adequately practical. Moreover, these regimens are associated with considerable complications. Designing tolerogenic therapeutic strategies based on stem cells, immune cells, and their extracellular vesicles (EVs) seems to open a promising path to managing autoimmune diseases’ vast burden. Mesenchymal stem/stromal cells (MSCs), dendritic cells, and regulatory T cells (Tregs) are the main cell types applied to restore a tolerogenic immune status; MSCs play a more beneficial role due to their amenable properties and extensive cross-talks with different immune cells. With existing concerns about the employment of cells, new cell-free therapeutic paradigms, such as EV-based therapies, are gaining attention in this field. Additionally, EVs’ unique properties have made them to be known as smart immunomodulators and are considered as a potential substitute for cell therapy. This review provides an overview of the advantages and disadvantages of cell-based and EV-based methods for treating autoimmune diseases. The study also presents an outlook on the future of EVs to be implemented in clinics for autoimmune patients.

show abstract

“…Although sEVs have been recognized as the potential candidates in clinical applications and have been explored widely, the low yield, inadequate therapeutic effect, and targeting efficiency of sEVs are still stumbling blocks of native sEVs for large-scale clinical applications. Very recently, a workshop, named “massivEVs”, organized by the International Society for Extracellular Vesicles (ISEV), “The Extracellular Vesicle Foundry” (evFOUNDRY), and “Extracellular vesicles from a natural source for tailor-made nanomaterials” (VES4US) also expressed serious concern on the large-scale production of sEVs from sources, upstream and downstream technologies, validation, standardization, and regulation ( Paolini et al, 2022 ). Up to now, numerous ingenious strategies have been developed to conquer those bottlenecks, promoting the application of sEVs in clinical practice.…”

Section: Scalable Production and Engineering Of Sevsmentioning

confidence: 99%

Small extracellular vesicles from mesenchymal stem cells: A potential Weapon for chronic non-healing wound treatment

Wei

Liu

et al. 2023

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Chronic non-healing wounds have posed a severe threat to patients mentally and physically. Behavior dysregulation of remaining cells at wound sites is recognized as the chief culprit to destroy healing process and hinders wound healing. Therefore, regulating and restoring normal cellular behavior is the core of chronic non-healing wound treatment. In recent years, the therapy with mesenchymal stem cells (MSCs) has become a promising option for chronic wound healing and the efficacy has increasingly been attributed to their exocrine functions. Small extracellular vesicles derived from MSCs (MSC-sEVs) are reported to benefit almost all stages of wound healing by regulating the cellular behavior to participate in the process of inflammatory response, angiogenesis, re-epithelization, and scarless healing. Here, we describe the characteristics of MSC-sEVs and discuss their therapeutic potential in chronic wound treatment. Additionally, we also provide an overview of the application avenues of MSC-sEVs in wound treatment. Finally, we summarize strategies for large-scale production and engineering of MSC-sEVs. This review may possibly provide meaningful guidance for chronic wound treatment with MSC-sEVs.

show abstract

Large‐scale production of extracellular vesicles: Report on the “massivEVs” ISEV workshop

Cited by 36 publications

References 55 publications

A novel approach for large-scale manufacturing of small extracellular vesicles from bone marrow-derived mesenchymal stromal cells using a hollow fiber bioreactor

A novel approach for large-scale manufacturing of small extracellular vesicles from bone marrow-derived mesenchymal stromal cells using a hollow fiber bioreactor

Extracellular vesicles and their cells of origin: Open issues in autoimmune diseases

Small extracellular vesicles from mesenchymal stem cells: A potential Weapon for chronic non-healing wound treatment

Contact Info

Product

Resources

About