Upscaling human mesenchymal stromal cell production in a novel vertical-wheel bioreactor enhances extracellular vesicle secretion and cargo profile.

Autor:	Jeske R; Department of Chemical and Biomedical Engineering, FAMU-FSU College of Engineering, Florida State University, Tallahassee, FL, USA., Liu C; Department of Chemical and Biomedical Engineering, FAMU-FSU College of Engineering, Florida State University, Tallahassee, FL, USA., Duke L; Department of Biomedical Sciences, College of Medicine, Florida State University, Tallahassee, FL, USA., Canonicco Castro ML; Department of Chemical and Biomedical Engineering, FAMU-FSU College of Engineering, Florida State University, Tallahassee, FL, USA., Muok L; Department of Chemical and Biomedical Engineering, FAMU-FSU College of Engineering, Florida State University, Tallahassee, FL, USA., Arthur P; College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, Tallahassee, FL, USA., Singh M; College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, Tallahassee, FL, USA., Jung S; PBS Biotech Inc., Camarillo, CA, USA., Sun L; Department of Chemical and Biomedical Engineering, FAMU-FSU College of Engineering, Florida State University, Tallahassee, FL, USA.; Department of Biomedical Sciences, College of Medicine, Florida State University, Tallahassee, FL, USA., Li Y; Department of Chemical and Biomedical Engineering, FAMU-FSU College of Engineering, Florida State University, Tallahassee, FL, USA.
Jazyk:	angličtina
Zdroj:	Bioactive materials [Bioact Mater] 2022 Aug 02; Vol. 25, pp. 732-747. Date of Electronic Publication: 2022 Aug 02 (Print Publication: 2023).
DOI:	10.1016/j.bioactmat.2022.07.004
Abstrakt:	Human mesenchymal stromal cells (hMSCs) are mechanically sensitive undergoing phenotypic alterations when subjected to shear stress, cell aggregation, and substrate changes encountered in 3D dynamic bioreactor cultures. However, little is known about how bioreactor microenvironment affects the secretion and cargo profiles of hMSC-derived extracellular vesicles (EVs) including the subset, "exosomes", which contain therapeutic proteins, nucleic acids, and lipids from the parent cells. In this study, bone marrow-derived hMSCs were expanded on 3D Synthemax II microcarriers in the PBS mini 0.1L Vertical-Wheel bioreactor system under variable shear stress levels at 25, 40, and 64 RPM (0.1-0.3 dyn/cm 2 ). The bioreactor system promotes EV secretion from hMSCs by 2.5-fold and upregulates the expression of EV biogenesis markers and glycolysis genes compared to the static 2D culture. The microRNA cargo was also altered in the EVs from bioreactor culture including the upregulation of miR-10, 19a, 19b, 21, 132, and 377. EV protein cargo was characterized by proteomics analysis, showing upregulation of metabolic, autophagy and ROS-related proteins comparing with 2D cultured EVs. In addition, the scalability of the Vertical-Wheel bioreactor system was demonstrated in a 0.5L bioreactor, showing similar or better hMSC-EV secretion and cargo content compared to the 0.1L bioreactor. This study advances our understanding of bio-manufacturing of stem cell-derived EVs for applications in cell-free therapy towards treating neurological disorders such as ischemic stroke, Alzheimer's disease, and multiple sclerosis. Competing Interests: The authors declare no competing interests. (© 2022 The Authors.)
Databáze:	MEDLINE
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