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Article first published online: 16 NOV 2011
Copyright © 2011 AlphaMed Press
Volume 29, Issue 12, pages 1983–1994, December 2011
How to Cite
Yasuda, A., Tsuji, O., Shibata, S., Nori, S., Takano, M., Kobayashi, Y., Takahashi, Y., Fujiyoshi, K., Hara, C. M., Miyawaki, A., Okano, H. J., Toyama, Y., Nakamura, M. and Okano, H. (2011), Significance of Remyelination by Neural Stem/Progenitor Cells Transplanted into the Injured Spinal Cord. STEM CELLS, 29: 1983–1994. doi: 10.1002/stem.767
Author contributions: A.Y.: conception and design, experiments, data analysis and interpretation, manuscript writing, and final approval of manuscript; O.T.: conception and design, experiments, surgery, and data analysis and interpretation, S.S.: experiments and data analysis and interpretation, S.N.: experiments, surgery, and data interpretation; M.T.: experiments, surgery, and data analysis; Y.K. and Y.T.: experiments and data interpretation; K.F., C.M.H., A.M., H.J.O. and Y.T.: data analysis and interpretation; M.N. and H.O.: conception and design, data analysis and interpretation, manuscript writing, and final approval of manuscript.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLSEXPRESS October 25, 2011.
- Issue published online: 16 NOV 2011
- Article first published online: 16 NOV 2011
- Accepted manuscript online: 25 OCT 2011 03:19PM EST
- Manuscript Accepted: 5 OCT 2011
- Manuscript Received: 6 JUN 2011
- Program for the Promotion of Fundamental Studies in Health Sciences of the National Institute of Biomedical Innovation (NIBIO); a grant from the Uehara Memorial Foundation
- Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (JSPS) and the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT)
- Project for the Realization of Regenerative Medicine and support for the core institutes for iPSC research from MEXT
- “Funding Program for World-leading Innovative R&D on Science and Technology” to H.O. from the Ministry of Health, Labor, and Welfare
- General Insurance Association of Japan; Research Fellowships for Young Scientists from the Japan Society for the Promotion of Science
- Keio Gijuku Academic Development Funds
- Grant-in-aid for the Global COE program from MEXT to Keio University
Additional Supporting Information may be found in the online version of this article.
|STEM_767_sm_suppinfofig1.eps||14659K||Supplementary Figure 1 Neurosphere formation by shi-NS/PCs. (A), (B): shi-NS/PCs formed neurospheres indistinguishable from those of wt-NS/PCs. Scale bar = 200 μm. (C): ATP assay of shi- and wt-NS/PCs. There was no significant difference in the proliferation rate between the wt- and shi-NS/PCs (n = 32 assays. Each type of NS/PC was derived from four embryos with eight cultures for each embryo)).|
|STEM_767_sm_suppinfofig2.eps||1801K||Supplementary Figure 2 Representative motor-evoked potential (MEP) recordings in the control and transplanted mice during the experiment. MEP waves were not detected in any group immediately after injury or transplantation. Although MEP waves were detected in the shi-NS/PC and wt-NS/PC groups 7 weeks after injury, no waves were detected in the control group. Compound motor action potentials (CMAPs) obtained after sciatic nerve stimulation were observed at all time points and in all groups.|
|STEM_767_sm_suppinfofig3.eps||3995K||Supplementary Figure 3 shi-NS/PCs in an ICR mouse strain background did not survive in the injured spinal cord of C57BL/6J hosts, resulting in no functional recovery. Contusive SCI was induced by IH impactor (60 kdyn) and NS/PCs (5 × 105 cells/mouse; labeled with Venus fluorescent protein lentivirally) were transplanted into the lesion site 9 days after injury. (A): Immunohistological analysis using an anti-GFP antibody revealed that no shi-NS/PCs survived in the injured spinal cord. (B): On the other hand, wt-NS/PCs survived in the injured spinal cords of C57BL/6J mice as well as NOD/SCID mice. Scale bar = 500 ?m. (C): Mean BMS scores for each group over the 42-day recovery period. Although there was no significant difference in the BMS scores among the three groups until 21 days after injury, at day 28 and thereafter, the wt-NS/PC group showed significantly improved BMS scores compared with the other two groups (control, n = 4; shi-NS/PC group, n = 6; wt-NS/PC group, n = 4).|
|STEM_767_sm_suppinfofig4.eps||1520K||Supplementary Figure 4 Quantification of trophic factor mRNA levels produced by neurospheres from both NS/PC genotypes. Trophic factor gene expression plots of the value 2(-Δ CT) with a three-fold difference are displayed, with shi-NS/PCs (y-axis) and wt-NS/PCs (x-axis). Sixty-four factors (see Supplementary Table 1 are dotted between the lines indicating a three-fold difference (n = 3 each). mRNAs for ciliary neurotrophic factor (CNTF), growth differentiation factor 11 (GDF11), platelet derived growth factor A (PDGFA) and vascular endothelial growth factor B (VEGFB) were expressed at relatively high levels by both the shi- and wt-NS/PC-derived neurospheres. Although every analysis of wt-NS/PC-derived neurospheres detected MBP mRNA expression, it was never detected in the analyses of shi-NS/PC-derived neurospheres, confirming their identity as MBP-deficient shiverer mutant cells.|
|STEM_767_sm_suppinfotab1.doc||467K||Supplementary Table 1. The primers used in this study are shown in this table. All primers were manufactured by and obtained from Applied Biosystems.|
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