September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Signaling factors secretion by 2D mesenchymal stem cells (MSCs) and spheroid MSCs and microvesicles derived from them cocultured with retinal ganglion cell line
Author Affiliations & Notes
  • Lili Xie
    Ophthalmology, The 2nd Xiangya Hospital,Central South University, Changsha, Hunan, China
  • Bing Jiang
    Ophthalmology, The 2nd Xiangya Hospital,Central South University, Changsha, Hunan, China
  • Mao Mao
    Univ of California, SF Sch of Med, San Francisco, California, United States
  • Footnotes
    Commercial Relationships   Lili Xie, None; Bing Jiang, None; Mao Mao, None
  • Footnotes
    Support  NSFC 81371012 to Bing Jiang
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 2570. doi:
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      Lili Xie, Bing Jiang, Mao Mao; Signaling factors secretion by 2D mesenchymal stem cells (MSCs) and spheroid MSCs and microvesicles derived from them cocultured with retinal ganglion cell line. Invest. Ophthalmol. Vis. Sci. 2016;57(12):2570.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Mesenchymal stem cells (MSCs) have been shown to be a potential treatment for degenerative eye diseases. In this study we aim to identify and compare levels of signaling factors secreted by MSCs cultured in 2D monolayer (2D-MSCs) or in spheroids (spheoird MSCs). To explore potential non cell-based therapies, we also treated retinal ganglion cells (RGC-5) with MSC derived microvesicles (MVs) and analyzed levels of secreted signaling factors.

Methods : We seeded 2D-MSCs, spheroid MSCs and cells isolated by trypsinization of spheoird MSCs at equal cell density and collected conditioned medium after 24hrs. Hanging drop protocol was used to prepare spheroid MSCs. We isolated microvesicles (MVs) from conditioned medium of MSCs cultured in those three conditions and co-cultured them with RGC-5 for different time period. Levels of 51 cytokines in conditioned medium of MSCs or MV treated RGC-5 were qualified with a high throughput bead-based assay.

Results : We found that IL-8, IL-6 and CXCL1 were the top three abundant cytokines secreted by MSCs. In addition, we found that compared to 2D-MSCs, protein levels of 12 cytokines and a cytokine receptor, IL-2Rα were significantly increased in conditioned medium from spheroid MSCs. Finally, we found increased levels of SCGF-β, VEGF and LIF in conditioned medium of RGC5 co-cultured with MVs derived from spheroid MSCs.

Conclusions : 3D suspension culture model improves the ability of MSCs to secrete signal factors responsible for anti-inflammation, cell differentiation and cell survival comparing to traditional 2D monolayer culture. SCGF-β, VEGF and LIF can be valuable targets for the progress of acellular grafts for application in RGC regenerative therapies.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

 

MSCs as spheroids with 25×103 cells/drop)

MSCs as spheroids with 25×103 cells/drop)

 

microvesicles derived from spheroid MSCs

microvesicles derived from spheroid MSCs

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