Exosomes Produced from 3D Cultures of MSCs by Tangential Flow Filtration Show Higher Yield and Improved Activity
Open accessAbstract
Exosomes can deliver therapeutic RNAs to neurons. The composition and the safety profile of exosomes depend on the type of the exosome-producing cell. Mesenchymal stem cells are considered to be an attractive cell type for therapeutic exosome production. However, scalable methods to isolate and manufacture exosomes from mesenchymal stem cells are lacking, a limitation to the clinical translation of exosome technology. We evaluate mesenchymal stem cells from different sources and find that umbilical cord-derived mesenchymal stem cells produce the highest exosome yield. To optimize exosome production, we cultivate umbilical cord-derived mesenchymal stem cells in scalable microcarrier-based three-dimensional (3D) cultures. In combination with the conventional differential ultracentrifugation, 3D culture yields 20-fold more exosomes (3D-UC-exosomes) than two-dimensional cultures (2D-UC-exosomes). Tangential flow filtration (TFF) in combination with 3D mesenchymal stem cell cultures further improves the yield of exosomes (3D-TFF-exosomes) 7-fold over 3D-UC-exosomes. 3D-TFF-exosomes are seven times more potent in small interfering RNA (siRNA) transfer to neurons compared with 2D-UC-exosomes. Microcarrier-based 3D culture and TFF allow scalable production of biologically active exosomes from mesenchymal stem cells. These findings lift a major roadblock for the clinical utility of mesenchymal stem cell exosomes.
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UC-MSCs secrete more Exos than BM- and AD-MSCs.
“It has been reported that UC-MSCs secrete more Exos than BM- and AD-MSCs [321].”
From Mechanisms, efficacy, and future perspectives of cellular-based therapies for liver fibrosis/cirrhosis: focusing on mesenchymal stromal cells (2025) · cites this paper as [321]
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“It has been reported that UC-MSCs secrete more Exos than BM-and AD-MSCs [321].”
- Authors
- Haraszti, R. A., Miller, R., Stoppato, M., Sere, Y. Y., Coles, A., Didiot, M.-C., Wollacott, R., Sapp, E., Dubuke, M. L., Li, X., Shaffer, S. A., DiFiglia, M., Wang, Y., Aronin, N., & Khvorova, A.
- Affiliations
- University of Massachusetts Chan Medical School (US)
- Massachusetts General Hospital (US)
- Corresponding author
- Khvorova, Anastasia
- ORCID
- Venue
- Molecular Therapy
- Publisher
- Elsevier BV
- Published
- 2018-12
- Volume
- 26
- Issue
- 12
- Pages
- 2838-2847
- ISSN
- 1525-0016 (ISSN-L)
- PMC
- PMC6277553
- OpenAlex
- W2892744778
- Cited by
- 567
- References
- 5
- Citations by year
- 2026: 83 · 2025: 100 · 2024: 97 · 2023: 86 · 2022: 86 · 2021: 53 · 2020: 45 · 2019: 17
- FWCI
- 10.23
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Molecular Therapy (publishedVersion, cc-by-nc-nd)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- The Journal of the American Medical Association (JAMA) Network (American Medical Association) (submittedVersion)
- Funders
- NIH (TR 000888 05, RO1GM10880304, RO1NS10402201, S10 OD020012)
- UMass (UL1 TR000161)
- CHDI Foundation (A-6119, JSC A6367)
- Huntington’s Disease Society of America Postdoctoral Fellowship
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
- Health Sciences › Medicine › Genetics › Mesenchymal stem cell research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- exosomes, mesenchymal stem cells, umbilical cord-derived stem cells, cross-flow filtration, exosome yield, exosome isolation, differential ultracentrifugation
- MeSH terms
- Animals, Cell Differentiation, Cells, Cultured, Female, Neurons, Umbilical Cord, Spheroids, Cellular, Cell Culture Techniques, Proteome, Gene Silencing, RNA, Small Interfering, Mice, Exosomes, Mesenchymal Stem Cells
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