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Version of Record online: 22 MAY 2013
Copyright © 2013 AlphaMed Press
Volume 31, Issue 6, pages 1136–1148, June 2013
How to Cite
Manuguerra-GagnÉ, R., Boulos, P. R., Ammar, A., Leblond, F. A., Krosl, G., Pichette, V., Lesk, M. R. and Roy, D.-C. (2013), Transplantation of Mesenchymal Stem Cells Promotes Tissue Regeneration in a Glaucoma Model Through Laser-Induced Paracrine Factor Secretion and Progenitor Cell Recruitment. STEM CELLS, 31: 1136–1148. doi: 10.1002/stem.1364
Author contributions: R.M.-G.: conception and design, collection and/or assembly of data, data analysis and interpretation, and manuscript writing; P.R.B. and V.P.: conception and design; A.A. and F.A.L.: conception and design and collection and/or assembly of data; G.K.: conception and design and manuscript writing; M.R.L. and D.-C.R.: conception and design, administrative support, financial support, manuscript writing.
Disclosure of potential conflicts of interest is found at the end of this article.
first published online in STEM CELLS EXPRESS February 4, 2013.
- Issue online: 22 MAY 2013
- Version of Record online: 22 MAY 2013
- Accepted manuscript online: 14 MAR 2013 12:00AM EST
- Manuscript Accepted: 5 FEB 2013
- Manuscript Received: 7 JUL 2012
- Glaucoma Research Society of Canada
- Thécell Network - Fonds Recherche Quebec - Sante (FRQS)
- Fonds de Recherche en Ophtalmologie de l'Université de Montréal
- Fonds de recherche Québec - Santé
- Canadian Institutes for Health Research
Additional Supporting Information may be found in the online version of this article.
|sc-12-0633_sm_SupplFigure1.pdf||1873K||Supplementary Figure 1. BMMC Characterization. BMMC population was characterized for hematopoietic cells and MSC. A) Using CD45 and CD14 hematopoietic marker, we identified 65% of non hematopoietic cells after 2 passages in vitro. Gated cells were positive for CD73 (B) CD90 (C) and CD105 (D). Selected CD45-CD14 negative cells were plated in adipogenic (E) and osteogenic (F) media, were allowed to differentiate and compared to non treated MSC controls. Representative images of flow cytometry analysis and MSC differentiation. Scale bar = 50 μm|
|sc-12-0633_sm_SupplFigure2.pdf||1876K||Supplementary Figure 2. MSC do not transdifferentiate into TM cells in vitro. A) TM cells and MSC before and after 1 week co-culture with TM cells were assessed for the presence of laminin, fibronectin, aquaporin-1 and Pax6 by flow cytometry. Fluorescence intensity was measured on cell tracker positive MSC (dot plot). B) Compilation of mean fluorescence intensity for each antigen in TM cells, MSC alone and MSC co-cultured with TM. (Mean±SEM; n=3 from 3 distinct experiments). Significance: *p<0.05, ***p<0.001.|
|sc-12-0633_sm_SupplFigure3.pdf||1570K||Supplementary Figure 3. MSC phagocytosis by ocular microglia. Pictures are representative images of day 2 post laser damage tissue sections that were stained with microglia marker F 4/80 after A) injection of saline solution or B) injection with CFSE+ MSC. Scale bar = 50 μm|
|sc-12-0633_sm_SupplFigure4.tif||1261K||Supplementary Figure 4. MSC paracrine factors promote restoration of TM function. Aqueous humor clearance was evaluated by measuring peripheral blood radioactive tracer levels in counts per minutes (CPM) at different time points after injection of radiotracer in normal eyes (black line, n=3), laser treated eyes (blue line, n=3) and low oxygen CM treated eyes (red line, n=3). Results are expressed as mean±SEM of 2 independent experiments. Statistical significance was observed in low O2 CM exposed eyes versus laser only and normal controls: *p<0.05|
|sc-12-0633_sm_SupplFigure5.pdf||1629K||Supplementary Figure 5. Decreased IOP allows for better survival of retinal cells Histological sections of the retinal area from a rat eye stained by TUNEL assay. Pictures are representative images of retinas from a A) untreated control B) Laser + normoxic CM treated eye, day 12 C) laser + low oxygen treated eye, day 12. D) Compilation of total number of TUNEL+ cells present in the retina 12 days after laser damage and CM injection. Cell numbers (mean±SEM) were determined by fluorescence microscopy analysis of sequential ocular sections (n=3 per time point). *p<0.05. Scale bar = 100 μm|
|sc-12-0633_sm_SupplFigure6.pdf||1656K||Supplementary Figure 6. Nestin expression and cellular proliferation in the damaged and intact areas. Graphs represent compilation of nestin (A-B) and Ki67 (C-D) expressing cells based on their location: Intact area (A-C) and damaged area (B-D). Three animals per group from 2 separate experiments (N=6). Numbers indicate the percentage of positive cells found in the area composed of the ciliary body, the TM and a limited portion of the iris.|
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