PMID- 35986305
OWN - NLM
STAT- MEDLINE
DCOM- 20220823
LR  - 20240516
IS  - 1757-6512 (Electronic)
IS  - 1757-6512 (Linking)
VI  - 13
IP  - 1
DP  - 2022 Aug 19
TI  - Bone marrow mesenchymal stromal cells in a 3D system produce higher concentration 
      of extracellular vesicles (EVs) with increased complexity and enhanced neuronal 
      growth properties.
PG  - 425
LID - 10.1186/s13287-022-03128-z [doi]
LID - 425
AB  - PURPOSE: Extracellular vesicles (EVs) derived from mesenchymal stromal cells 
      (MSCs) have been demonstrated to possess great potential in preclinical models. 
      An efficient biomanufacturing platform is necessary for scale up production for 
      clinical therapeutic applications. The aim of this study is to investigate the 
      potential differences in neuro-regenerative properties of MSC-derived EVs 
      generated in 2D versus 3D culture systems. METHOD: Human bone marrow MSCs 
      (BM-MSCs) were cultured in 2D monolayer and 3D bioreactor systems. EVs were 
      isolated using ultracentrifugation followed by size and concentration 
      measurements utilizing dynamic light scattering (NanoSight) and by fluorescence 
      staining (ExoView). Mouse trigeminal ganglia (TG) neurons were isolated from 
      BALB/c mice and cultured in the presence or absence of EVs derived from 2D or 3D 
      culture systems. Neuronal growth and morphology were monitored over 5 days 
      followed by immunostaining for beta3 tubulin. Confocal images were analyzed by 
      Neurolucida software to obtain the density and length of the neurites. RESULTS: 
      The NanoSight tracking analysis revealed a remarkable increase (24-fold change) 
      in the concentration of EVs obtained from the 3D versus 2D culture condition. 
      ExoView analysis showed a significantly higher concentration of CD63, CD81, and 
      CD9 markers in the EVs derived from 3D versus 2D conditions. Furthermore, a 
      notable shift toward a more heterogeneous phenotype was observed in the 
      3D-derived EVs compared to those from 2D culture systems. EVs derived from both 
      culture conditions remarkably induced neurite growth and elongation after 5 days 
      in culture compared to untreated control. Neurolucida analysis of the 
      immunostaining images (beta3 tubulin) showed a significant increase in neurite 
      length in TG neurons treated with 3D- versus 2D-derived EVs (3301.5 mum vs. 
      1860.5 mum, P < 0.05). Finally, Sholl analysis demonstrated a significant increase 
      in complexity of the neuronal growth in neurons treated with 3D- versus 
      2D-derived EVs (P < 0.05). CONCLUSION: This study highlights considerable 
      differences in EVs obtained from different culture microenvironments, which could 
      have implications for their therapeutic effects and potency. The 3D culture 
      system seems to provide a preferred environment that modulates the paracrine 
      function of the cells and the release of a higher number of EVs with enhanced 
      biophysical properties and functions in the context of neurite elongation and 
      growth.
CI  - (c) 2022. The Author(s).
FAU - Jalilian, Elmira
AU  - Jalilian E
AUID- ORCID: 0000-0003-4121-0517
AD  - Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, 
      University of Illinois at Chicago, 1855 W. Taylor Street, MC 648, Chicago, IL, 
      60612, USA. jalilian@uic.edu.
AD  - Richard and Loan Hill Department of Bioengineering, University of Illinois at 
      Chicago, Chicago, IL, USA. jalilian@uic.edu.
FAU - Massoumi, Hamed
AU  - Massoumi H
AD  - Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, 
      University of Illinois at Chicago, 1855 W. Taylor Street, MC 648, Chicago, IL, 
      60612, USA.
AD  - Richard and Loan Hill Department of Bioengineering, University of Illinois at 
      Chicago, Chicago, IL, USA.
FAU - Bigit, Bianca
AU  - Bigit B
AD  - Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, 
      University of Illinois at Chicago, 1855 W. Taylor Street, MC 648, Chicago, IL, 
      60612, USA.
FAU - Amin, Sohil
AU  - Amin S
AD  - Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, 
      University of Illinois at Chicago, 1855 W. Taylor Street, MC 648, Chicago, IL, 
      60612, USA.
FAU - Katz, Eitan A
AU  - Katz EA
AD  - Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, 
      University of Illinois at Chicago, 1855 W. Taylor Street, MC 648, Chicago, IL, 
      60612, USA.
FAU - Guaiquil, Victor H
AU  - Guaiquil VH
AD  - Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, 
      University of Illinois at Chicago, 1855 W. Taylor Street, MC 648, Chicago, IL, 
      60612, USA.
FAU - Anwar, Khandaker N
AU  - Anwar KN
AD  - Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, 
      University of Illinois at Chicago, 1855 W. Taylor Street, MC 648, Chicago, IL, 
      60612, USA.
FAU - Hematti, Peiman
AU  - Hematti P
AD  - Department of Medicine, Hematology/Oncology Division, University of 
      Wisconsin-Madison, School of Medicine and Public Health, Madison, WI, 53705, USA.
FAU - Rosenblatt, Mark I
AU  - Rosenblatt MI
AD  - Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, 
      University of Illinois at Chicago, 1855 W. Taylor Street, MC 648, Chicago, IL, 
      60612, USA.
FAU - Djalilian, Ali R
AU  - Djalilian AR
AD  - Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, 
      University of Illinois at Chicago, 1855 W. Taylor Street, MC 648, Chicago, IL, 
      60612, USA. adjalili@uic.edu.
LA  - eng
GR  - EY031809/NH/NIH HHS/United States
GR  - UG3 EY031809/EY/NEI NIH HHS/United States
GR  - P30 EY001792/EY/NEI NIH HHS/United States
GR  - EY01792/NH/NIH HHS/United States
GR  - TR002002/NH/NIH HHS/United States
GR  - UH3 EY031809/EY/NEI NIH HHS/United States
GR  - KL2 TR002002/TR/NCATS NIH HHS/United States
GR  - R01 EY024349/EY/NEI NIH HHS/United States
GR  - EY024349/NH/NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20220819
PL  - England
TA  - Stem Cell Res Ther
JT  - Stem cell research & therapy
JID - 101527581
RN  - 0 (Tubulin)
SB  - IM
MH  - Animals
MH  - Bone Marrow
MH  - Bone Marrow Cells
MH  - *Extracellular Vesicles/physiology
MH  - Humans
MH  - *Mesenchymal Stem Cells
MH  - Mice
MH  - Tubulin
PMC - PMC9389821
OTO - NOTNLM
OT  - 2D culture
OT  - 3D culture
OT  - Bone marrow mesenchymal stromal cell
OT  - Exosomes
OT  - Extracellular vesicles
OT  - Neuronal growth
COIS- The authors declare that they have no competing interests.
EDAT- 2022/08/20 06:00
MHDA- 2022/08/24 06:00
PMCR- 2022/08/19
CRDT- 2022/08/19 23:36
PHST- 2022/06/14 00:00 [received]
PHST- 2022/08/08 00:00 [accepted]
PHST- 2022/08/19 23:36 [entrez]
PHST- 2022/08/20 06:00 [pubmed]
PHST- 2022/08/24 06:00 [medline]
PHST- 2022/08/19 00:00 [pmc-release]
AID - 10.1186/s13287-022-03128-z [pii]
AID - 3128 [pii]
AID - 10.1186/s13287-022-03128-z [doi]
PST - epublish
SO  - Stem Cell Res Ther. 2022 Aug 19;13(1):425. doi: 10.1186/s13287-022-03128-z.