1. Volume 18, Issue 10, Pages 1837-1845 (October 2010)
A Therapeutic Strategy for Choroidal Neovascularization Based on Recruitment of Mesenchymal Stem Cells to the Sites of Lesions 
Hui-Yuan Hou, Hong-Liang Liang, Yu-Sheng Wang, Zhao-Xia Zhang, Bai-Ren Wang, Yuan-Yuan Shi, Xiao Dong, Yan Cai 
Molecular Therapy 
Volume 18, Issue 10, Pages (October 2010)
DOI: /mt
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

2. Figure 1
Quantitative analysis of GFP-expressing mesenchymal stem cells (MSCs) in PB and BM by flow cytometry after laser photocoagulation and GFP-expressing MSC transplantation. (a) Representative data from three experiments are shown. (b) The quantity of GFP-expressing MSCs in BM increased obviously in the first 24 hours of the experiment and decreased rapidly thereafter. The number of MSCs in PB slightly increased on day 1 (error bars, SEM, n = 6). BM, bone marrow; GFP, green fluorescent protein; PB, peripheral blood.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

3. Figure 2
Representative choroidal flat mount preparations after laser photocoagulation and GFP-expressing mesenchymal stem cell (MSC) transplantation. Blood vessels were stained by rhodamine-conjugated agglutinin (red) in choroidal flat mounts. Panels show the recruitment of GFP-expressing MSCs (green) to the choroidal neovascularization (CNV) lesion (red) on days 1, 3, and 7, indicating a directional movement of the MSCs into the CNV lesions. On day 7, GFP-expressing MSCs in CNV participated in vascular structure formation (yellow). Bar = 50 µm. GFP, green fluorescent protein.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

4. Figure 3
Immunofluorescence staining of eye sections showing differentiated GFP-expressing mesenchymal stem cells (MSCs) in choroidal neovascularization (CNV) 1 week after CNV induction. The differentiation of MSCs in CNV was analyzed by cell maker staining. Representative confocal images show that MSCs (green) express (a) the mature vascular endothelial cell marker CD31 (red), (b) the vascular smooth muscle cell marker αSMA (red), (c) the macrophage marker F4/80 (red), (d) the fibroblast marker vimentin (red), and (e) the epithelial cell marker keratin (red). Blue: DAPI-stained nuclei. Bar = 20 µm. αSMA, α-smooth muscle actin; GFP, green fluorescent protein.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

5. Figure 4
Analysis of adenoviral transduction efficiency 24 hours after transduction and enzyme-linked immunosorbent assays of human pigment epithelial-derived factor (PEDF) released by adenoviral vectors expressing PEDF (AdPEDF)–transduced mesenchymal stem cells (MSCs). Representative inverted microscopy images show reporter green fluorescent protein (GFP) expression in MSCs (a, light microscope image; b, fluorescent image). (c) A representative flow cytometry analysis shows that 78.1% cells within the total cell population expressed GFP. (d) Human PEDF production by AdPEDF-transduced MSCs was detected in vitro during an 8-day experimental period. Maximum production was observed during the first 24 hours after infection (error bars, SEM, n = 9). Bar = 20 µm.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

6. Figure 5
Immunofluorescent staining and enzyme-linked immunosorbent assays (ELISAs) of human PEDF expression in the eyes of mice treated with adenoviral vectors expressing PEDF (AdPEDF)–transduced MSCs. (a) In eye sections of the AdPEDF-expressing mice, positive human PEDF staining (red) was found in AdPEDF-transduced MSCs (green) and the extracellular matrix nearby. Blue: DAPI-stained nuclei. (b) ELISAs show that the average production of human PEDF in eyes of the AdPEDF-expressing mice was relatively steady for 1 week (error bars, SEM, n = 6). Bar = 20 µm. GFP, green fluorescent protein; PEDF, pigment epithelial-derived factor.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

7. Figure 6
Hematoxylin and eosin (H&E) staining, choroidal flat mount, and quantitative analysis of choroidal neovascularization (CNV) severity 1 week after laser photocoagulation. (a) Representative H&E images show that CNVs from the AdPEDF group were encapsulated by pigmented cells (arrows), whereas CNVs in other groups were not. CNVs are indicated by dotted lines. The average thickness and diameter of CNVs from the AdPEDF group significantly decreased compared to other groups. No statistical difference was found among the other groups (error bars, SEM, n = 8). Bar = 20 µm. (b) The images of choroidal flat mounts show smaller surface areas of CNV (red) in the AdPEDF group. CNV area was significantly reduced in the AdPEDF group. No statistically significant difference was found among the other groups (error bars, SEM, n = 8). Bar = 50 µm. AdPEDF, adenoviral vectors expressing pigment epithelial-derived factor.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

8. Figure 7
Proliferation and migration assays of RPE cells cocultured with MSCs. (a) Migration of RPE cells was stimulated by the secretion of AdPEDF-transduced MSCs after coculture for 8 hours. (b) Three days after coculture, proliferation of RPE cells cocultured with AdPEDF-transduced MSCs was markedly enhanced. There was no statistically significant difference between the control group, the AdNull group, and the nontransduction group in both assays (error bars, SEM, n = 10). AdPEDF, adenoviral vectors expressing pigment epithelial-derived factor.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions