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Embryonic Stem Cells/Induced Pluripotent Stem Cells
Article first published online: 9 APR 2012
Copyright © 2012 AlphaMed Press
Volume 30, Issue 5, pages 865–875, May 2012
How to Cite
Kele, J., Andersson, E. R., Villaescusa, J. C., Cajanek, L., Parish, C. L., Bonilla, S., Toledo, E. M., Bryja, V., Rubin, J. S., Shimono, A. and Arenas, E. (2012), SFRP1 and SFRP2 Dose-Dependently Regulate Midbrain Dopamine Neuron Development In Vivo and in Embryonic Stem Cells. STEM CELLS, 30: 865–875. doi: 10.1002/stem.1049
Author contributions: J.K. and E.R.A.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing, and final approval of manuscript; J.C.V.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing, assembly of figures, and final approval of manuscript; L.C., V.B., and C.L.P.: collection and assembly of data and final approval of manuscript; S.B. and E.M.T.: collection of data and final approval of manuscript; J.S.R. and A.S.: provision of study material and final approval of manuscript; E.A.: conception and design, provision of study material, financial support, assembly of data, data analysis and interpretation, manuscript writing, and final approval of manuscript. J.K., E.R.A., and J.C.V. 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 January 30, 2012.
- Issue published online: 9 APR 2012
- Article first published online: 9 APR 2012
- Accepted manuscript online: 30 JAN 2012 03:50PM EST
- Manuscript Accepted: 29 DEC 2011
- Manuscript Received: 12 JUL 2011
- Swedish Research Council. Grant Numbers: DBRM, VR2008:2811, 3287
- European Union (Neurostemcell and Eurostemcell)
- Swedish Foundation for Strategic Research (INGVAR and DBRM)
- Norwegian Research Council
- Karolinska Institute
- National Health and Medical Research Council (NHMRC), Australia
- Swedish National Neuroscience Network
- FEBS Long-Term Fellowship
- EMBO Installation Grant and Ministry of Education, Youth and Sports of the Czech Republic. Grant Number: MSM0021622430
- NHMRC Career development Fellowship, Australia
- Ludwig Institute for Cancer Research Ltd., Stockholm, Sweden
- Department of Cellular and Molecular Biology, Karolinska Institute, Stockholm, Sweden
- Instituto de Biomedicina de Sevilla, Hospital Virgen del Rocio, Sevilla, Spain
- Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic
- Institute of Biophysics, Brno, Czech Republic
Additional Supporting Information may be found in the online version of this article.
|STEM_1049_sm_SuppFig1.pdf||102K||Supplementary Figure 1. Temporal pattern of expression of sFRP1-4 in the developing ventral and dorsal midbrain. (A-D) Quantitative RT PCR analysis was performed with rat ventral midbrain (vm) tissue obtained at embryonic days 10.5, 11.5, 13.5 and 15.5, and compared to dorsal midbrain (dm). Note that the initially high levels of sFRP1-3 mRNAs (A-C) are downregulated by the end of the neurogenic period. SFRP4 mRNA levels remained low and were not temporally regulated (D). *** p<0.001, ** p<0.01 and * p<0.05 compared to vm or dm at E10.5, respectively. One-way ANOVA, with Bonferroni's post-hoc test for multiple comparisons (N=3).|
|STEM_1049_sm_SuppFig2.tif||754K||Supplementary Figure 2. High concentrations of sFRP2 have no effect on basal or Wnt5ainduced DA differentiation of VM primary precursor cultures. (A) Rat E14.5 ventral midbrain cultures were treated for 3 days with vehicle (BSA Ctrl), sFRP1 or sFRP2 (5μg/ml) and analyzed for expression of beta III-tubulin, (TuJ1 - red) and tyrosine hydroxylase (TH - green) and counterstained with Hoechst (blue). (B) The number of TuJ1+ and TH+ neurons decreased in sFRP1-treated cultures. (C) Neither Wnt5a nor SFRP2 had any effect on the number of neurons. (D) While Wnt5a increased the number of TH+ cells, sFRP2 had no effect on the number of TH+ cells and did not modulate the effects of Wnt5a. * p<0.05 One-way ANOVA, with Bonferroni's post-hoc test for multiple comparisons (N=3).|
|STEM_1049_sm_SuppFig3.tif||1302K||Supplementary Figure 3. Proliferation in sFRP1−/−sFRP2−/− mice. (A) The number of A9-A10 VM TH+ cells was not altered in sFRP1−/−;sFRP2−/− embryos, compared to wild-type embryos at E14.5. (B) Quantification of Ki67 revealed no difference in the number of Ki67+ cells in WT and sFRP1−/−sFRP2−/− VM. (C) Stainings for Ki67 and Tuj1 of showed no detectable difference between sFRP1−/− and littermates sFRP1−/−;sFRP2−/− mice in the total number of neurons or cells in the cycle.|
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