AI Article Synopsis
- Human mesenchymal stromal cells (hMSCs) undergo significant changes in phenotype and extracellular vesicle (EV) profiles when cultured in dynamic 3D bioreactor systems, influenced by factors like shear stress and aggregation.
- The study found that using a Vertical-Wheel bioreactor increases EV secretion from hMSCs by 2.5 times compared to traditional 2D cultures, while also enhancing the expression of key genetic markers involved in EV production and metabolism.
- The findings suggest that optimizing bioreactor conditions can improve the therapeutic potential of hMSC-derived EVs, aiding in treatments for neurological disorders such as stroke and Alzheimer's disease.
Article Abstract
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). 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.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10087597 | PMC |
| http://dx.doi.org/10.1016/j.bioactmat.2022.07.004 | DOI Listing |
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Article Synopsis
- The study explores the use of suspension culturing in vertical wheel bioreactors to optimize stem cell expansion, addressing challenges in scalability and cell aggregation stability.
- Various media additives, including PEG and Heparin, were tested to improve pluripotency, expansion rate, and reduce aggregate fusion, resulting in a 40% shorter doubling time compared to traditional methods.
- The optimized solution maintained high pluripotency markers in two different cell lines, demonstrating that careful formulation and control of culture conditions can enhance stem cell growth and stability.
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