| Title: |
Scalable production of small extracellular vesicles derived from a human beta cell line cultured in stirred tank bioreactor with high yield and purity |
| Authors: |
Dauphin, Thibaud; Binet, Floriane; Pruvost, Quentin; Makula, Julien; Claire, Clémentine; Lalanne, Valerie; Hervé, Julie; Bach, Jean-Marie; Bosch, Steffi; Lieubeau, Blandine; de Beaurepaire, Laurence; Mosser, Mathilde |
| Contributors: |
Immuno-Endocrinologie Cellulaire et Moléculaire (IECM); École nationale vétérinaire, agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE); Bioproduction for Health and Innovative Therapy (B-FHIT); École nationale vétérinaire, agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-École nationale vétérinaire, agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE); ANR-21-CE18-0036,Bioprod-sEVforT1D,Bioproduction de vésicules extracellulaires thérapeutiques de cellules à insuline(2021) |
| Source: |
7th Congress of the FSEV; https://hal.science/hal-05052858; 7th Congress of the FSEV, Nov 2024, Strasbourg, France |
| Publisher Information: |
CCSD |
| Publication Year: |
2024 |
| Subject Terms: |
[SDV]Life Sciences [q-bio]; [SPI]Engineering Sciences [physics] |
| Subject Geographic: |
Strasbourg; France |
| Description: |
International audience ; FSEV2400067Scalable purification of small extracellular vesicles derived from a human cell line cultured in stirred tank bioreactor with high yield and purityT. Dauphin1, F. Binet1,2, Q. Pruvost1, J. Makula1, C. Claire1,2, V. Lalanne1,2, J. Herve1,2, J.M. Bach1, S. Bosch1, B. Lieubeau1,2, L. De Beaurepaire1,2, M. Mosser11ONIRIS, INRAE, IECM - NANTES (France), 2 ONIRIS, B-FHIT - NANTES (France)Background. Therapeutic applications of small extracellular vesicles (sEV) are gaining significant attention, but challenges remain in ensuring standardization and large-scale production. sEV are a heterogeneous population of particles smaller than 200 nm, with their composition and purity varying according to the cell culture conditions and the methods of separation. Commonly, sEV are produced from adherent cells cultured in static T-flask, and isolated by ultracentrifugation based-methods (UC). The use of stirred-tank bioreactors is crucial for process scalability but requires fine-tuning of hydrodynamic parameters across scales. Furthermore, UC methods have drawbacks, such as EV aggregation and manual workload. Therefore, it is essential to establish a downstream processes that integrated scalable technologies such as in-depth filtration, tangential flow filtration and low-pressure chromatography. In this study, we present the development of a scalable process for sEV production and purification.Material & Methods. The human beta cell line 1.4E7 was used as a model of anchorage-dependent cells. A process was established in which the cell culture in T-flask was replaced by spheroid culture in stirred tank systems. It was evaluated whether the best scale-up strategy for standardizing spheroid formation was to keep either the volumetric power input (P/V) or the tip speed constant in spinner flask. The impact of this up-scale process was assessed by analysis of spheroid morphology and viability. The process was then transferred to a fully controlled bioreactor. For EV isolation, the feasibility ... |
| Document Type: |
conference object; still image |
| Language: |
English |
| Availability: |
https://hal.science/hal-05052858; https://hal.science/hal-05052858v1/document; https://hal.science/hal-05052858v1/file/Poster-FSEV-VF.pdf |
| Rights: |
https://creativecommons.org/licenses/by-nc-nd/4.0/ ; info:eu-repo/semantics/OpenAccess |
| Accession Number: |
edsbas.8F28D90D |
| Database: |
BASE |