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Tissue-Specific Stem Cells
Version of Record online: 16 NOV 2011
Copyright © 2011 AlphaMed Press
Volume 29, Issue 12, pages 2030–2041, December 2011
How to Cite
Alfaro, L. A. S., Dick, S. A., Siegel, A. L., Anonuevo, A. S., McNagny, K. M., Megeney, L. A., Cornelison, D.D.W. and Rossi, F. M.V. (2011), CD34 Promotes Satellite Cell Motility and Entry into Proliferation to Facilitate Efficient Skeletal Muscle Regeneration. STEM CELLS, 29: 2030–2041. doi: 10.1002/stem.759
Author contributions: L.A.S.A.: conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing, and final approval of manuscript; S.A.D. and A.L.S.: conception and design, collection and/or assembly of data, data analysis and interpretation and final approval of manuscript; A.S.A.: collection and/or assembly of data and final approval of manuscript; K.M.M.: conception and design, provision of study material, data interpretation, manuscript writing, and final approval of manuscript; L.A.M.: conception and design, data interpretation, and final approval of manuscript; D.D.W.C.: conception and design, collection and/or assembly of data, data analysis and interpretation, manuscript writing, final approval of manuscript, and financial support; F.M.V.R.: conception and design, data interpretation, manuscript writing, final approval of manuscript, and financial support. S.A.D. and A.L.S. contributed equally to this article.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLSEXPRESS October 13, 2011.
- Issue online: 16 NOV 2011
- Version of Record online: 16 NOV 2011
- Accepted manuscript online: 13 OCT 2011 04:18PM EST
- Manuscript Accepted: 23 SEP 2011
- Manuscript Received: 11 JUL 2011
- CIHR. Grant Number: MOP-82864
- National Institute of Health
- NIH. Grant Number: 1R21AR056814-01
- Michael Smith Foundation for Health Research
- Natural Sciences and Engineering Research Council of Canada. Grant Numbers: NSERC, PGSD2-362406-2008
- Michael Smith Foundation for Health Research (MSFHR, ST-JGS-062(06-1)BM)
Additional Supporting Information may be found in the online version of this article.
|STEM_759_sm_suppinfofig1.tif||8291K||Supplemental Figure S1. Isolation of myogenic progenitor cells using FACS. (A) Plots showing gating used for MPC isolation by flow cytometry. MPCs were isolated by gating for the nucleated, live cell fraction (Hoechst+ and PI-, respectively), followed by exclusion of hematopoietic and endothelial cells (assessed by CD45- and CD31-, respectively), leaving MPCs exclusively within the Sca1- Alpha7 integrin+ population.|
|STEM_759_sm_suppinfofig2.tif||2050K||Supplemental Figure S2. Quantification of absolute values of sorted WT and Cd34?/? MPCs in multiple experiments. (A) LacZ+ MPCs were sorted and used as internal controls injected with WT/GFP+ or Cd34−/−/GFP+ in three independent experiments. Total numbers of LacZ+ fibers were determined. Error bars represent ± SEM for n = 3-5 mice per experiment. (B) Total numbers of GFP+ fibers from WT or Cd34−/− transplanted MPCs were determined in four independent experiments. Error bars represent ± SEM for n = 3-5 mice per experiment. (C) Frequency of Myf5/LacZ+ cells in WT and Cd34−/& minus; sorted MPCs in four independent experiments. Error bars represent ± SEM for n = 3 mice per experiment.|
|STEM_759_sm_suppinfofig3.tif||6527K||Supplemental Figure S3. Cd34−/− satellite cells efficiently cross the basal lamina on cultured single fibers. (A) WT and Cd34−/− single fibers were cultured and harvested at 0, 24, 48, and 72 hours post-culture. Immunofluorescent staining for satellite cells (Pax7) and the basal lamina (laminin) was performed and the location of satellite cells relative to the basal lamina was determined (Pax7, red; laminin, green). Representative images are shown. Bar graphs on the right show the relative proportion of satellite cells below or above the basal lamina in WT and Cd34−/− groups, n = 3 animals. Scale bar = 25 μm.|
|STEM_759_sm_suppinfovideo1.mpg||1799K||Supplemental online video S1. Live videomicroscopy imaging of WT satellite cell motility. Live imaging of satellite cells from isolated WT fibers was performed during the 24-48 hours period in culture.|
|STEM_759_sm_suppinfovideo2.mpg||1564K||Supplemental online video S2. Live videomicroscopy imaging of Cd34−/− satellite cell motility. Live imaging of satellite cells from isolated Cd34−/− fibers was performed during the 24-48 hours period in culture. Representative videos are shown.|
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