(176g) 3D Dynamic Culture Enhances Extracellular Vesicle Production with Altered Biogenesis and Cargo Contents in Human Mesenchymal Stem Cells | AIChE

(176g) 3D Dynamic Culture Enhances Extracellular Vesicle Production with Altered Biogenesis and Cargo Contents in Human Mesenchymal Stem Cells

Authors 

Yuan, X. - Presenter, FAMU-FSU College of Engineering
Nkosi, D., The Florida State University
Fu, Q., Florida State Univeristy
Liu, Y., Florida State University
Sun, L., The Florida State University
Meckes, D., Florida State University
Ma, T., FAMU-FSU College of Engineering

3D
Dynamic Culture Enhances Extracellular Vesicle Production with Altered
Biogenesis and Cargo Contents in Human Mesenchymal Stem Cells

Xuegang Yuan1, 2, Dingani Nkosi3, Qin Fu1, Yuan Liu1,
Richard Jeske1, Li Sun3, David Meckes3 and
Teng Ma1,2

1Chemical
and Biomedical Engineering; 2The National High Magnetic Field
Laboratory; 3Department of Biomedical Sciences, College of Medicine

The Florida State University, Tallahassee,
Florida, USA

Key words: Human mesenchymal stem cell; 3D aggregation; extracellular
vesicles; exosome; immunomodulation; manufacturing

Abstract:

Human mesenchymal stem cells (hMSCs) have demonstrated significant therapeutic potential and
are major candidate in cell therapy in a wide range of diseases including stroke,
multiple sclerosis, cardiovascular, graft versus host disease.  The clinical efficacy of hMSCs is mainly attributed to the paracrine effects thorough
hMSC secretome.  Among various components of secretome, extracellular vehicles (EVs) are cell-derived
vesicles responsible for cell-cell communication and regulation of cellular
events.  hMSCs
derived EVs, including microvesicles (MVs) and
exosomes, contain various cargos such as DNA, proteins, miRNA and organelles, exerting
paracrine effect and cell-cell communication to facilitate immunomodulatory
activity [1]. With several clinical trials ongoing, hMSC
derived EVs possess the potential as cell-free therapeutic strategy in
regenerative medicine, with feasibility of biomanufacturing. However, scale up
of hMSC derived EVs has become a major challenge for
clinical research. Studies have shown low efficiency of EVs secretion under
conventional monolayer culture. Unstable EV properties with regards to the cargo
and functional difference are also reported and may be due to the heterogeneity
and senescence during culture expansion. In current study, we employ dynamic 3D
aggregation culture of hMSCs to improve the
bioprocessing strategy of EVs production. hMSCs were
cultured as 3D aggregates under rocking motion to facilitate aggregation [2].
EVs production was significantly improved under 3D dynamic culture of hMSCs. Moreover, 3D hMSC-EVs also
exhibited smaller size compared to 2D hMSC-EV from nanoparticle
tracking analysis (NTA), with higher exosome enrichment. Our results also
demonstrated enhanced biogenesis of EVs in 3D hMSC
aggregates. Proteomic and transcriptomic analysis were also conducted to
demonstrate the difference of cargos between 3D dynamic culture and monolayer
culture. For functional evaluation, 3D hMSC aggregate
derived EVs has similar effect of immunomodulation compared to EVs derived from
hMSCs under monolayer culture. Moreover, 3D hMSC aggregate derived EVs exhibited enhanced
anti-senescence when cultured with hMSCs with
replicative senescence. Together, our study provides a promising bioprocess for
hMSCs derived EVs production with enhanced
therapeutic potentials compared to conventional monolayer culture.

Reference:

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D. ExtraPEG: A Polyethylene Glycol-Based Method for
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2. Tsai AC., Liu Y., Yuan X., Chella
R., Ma T. Aggregation Kinetics of Human Mesenchymal Stem Cells Under Wave
Motion. Biotechnol J. 2017;
12(5).