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Mol Ther. 2018 Sep 22. pii: S1525-0016(18)30456-8. doi: 10.1016/j.ymthe.2018.09.015. [Epub ahead of print]

Exosomes Produced from 3D Cultures of MSCs by Tangential Flow Filtration Show Higher Yield and Improved Activity.

Author information

1
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA.
2
Department of Medicine, University of Massachusetts Medical School, Worcester, MA, USA.
3
MassBiologics, Boston, MA, USA.
4
Mass General Institute for Neurodegenerative Disease, Boston, MA, USA.
5
Mass Spectrometry Facility, University of Massachusetts Medical School, Shrewsbury, MA, USA.
6
Department of Medicine, University of Massachusetts Medical School, Worcester, MA, USA. Electronic address: neil.aronin@umassmed.edu.
7
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA; Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, USA. Electronic address: anastasia.khvorova@umassmed.edu.

Abstract

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.

KEYWORDS:

exosomes; mesenchymal stem cell; tangential flow filtration

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