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Stem Cell Technology: Epigenetics, Genomics, Proteomics, and Metabonomics
Version of Record online: 19 APR 2011
Copyright © 2011 AlphaMed Press
Volume 29, Issue 5, pages 836–846, May 2011
How to Cite
Kondo, T., Matsuoka, A. J., Shimomura, A., Koehler, K. R., Chan, R. J., Miller, J. M., Srour, E. F. and Hashino, E. (2011), Wnt Signaling Promotes Neuronal Differentiation from Mesenchymal Stem Cells Through Activation of Tlx3. STEM CELLS, 29: 836–846. doi: 10.1002/stem.624
Author contributions: T.K.: conception and design, collection and/or assembly of data; data analysis and interpretation, manuscript writing; A.J.M.: conception and design, collection and/or assembly of data, data analysis and interpretation; A.S.: collection and/or assembly of data, data analysis and interpretation, manuscript writing; K.R.K.: collection and/or assembly of data, data analysis and interpretation; R.J.C.: collection and/or assembly of data, data analysis and interpretation, final approval of manuscript; J.M.M.: data analysis and interpretation, final approval of manuscript; E.F.S.: collection and/or assembly of data, data analysis and interpretation, final approval of manuscript; E.H.: conception and design, financial support, data analysis and interpretation, manuscript writing, final approval of manuscript.
First published online in STEM CELLS EXPRESS March 3, 2011.
Disclosure of potential conflicts of interest is found at the end of this article.
- Issue online: 19 APR 2011
- Version of Record online: 19 APR 2011
- Accepted manuscript online: 3 MAR 2011 04:48PM EST
- Manuscript Accepted: 1 FEB 2011
- Manuscript Received: 7 SEP 2010
- National Institutes of Health. Grant Numbers: R01 DC007390, RC1 DC010706
- Ruth and Lynn Townsend Professorship in Communication Disorders to the University of Michigan
Additional Supporting Information may be found in the online version of this article.
|STEM_624_sm_suppinfoFigs.pdf||220K||Supplemental Figure 1 Modifications of Alzet osmotic pumps. Polyimide tubing was inserted into a vinyl tubing and secured with silicon rubber. A small drop of silicone rubber placed 0.3 mm from the fine tip kept the cannula from being introduced too far into the scala tympani and helped seal the infusion hole in the cochlea. Supplemental Figure 2 Wnt3a induces expression of sensory neuron markers in neurally-induced MSCs. qRT-PCR analysis for Ngn1, NeuroD, Brn3a and Tlx3 in MSCs grown in neural induction medium containing recombinant human Wnt3a (1-200 ng/mL) for 7 days. Values are mean ± SD. Supplemental Figure 3 Sensory neuron marker genes are not induced by Wnt1 in MSCs without neural induction. RT-PCR analysis for Ngn1, NeuroD, Brn3a and L27 (internal control) in MSCs grown in maintenance medium containing recombinant human Wnt1 (100 ng/mL) for 3 (d3) or 7 days (d7). PC; E10 otocyst cDNA, NC; no cDNA. Supplemental Figure 4 Wnt1 induces expression of AMPA receptors in neurally-induced MSCs. (A) RT-PCR analysis for AMPA receptors (GluR1,-2,-3 and -4) in MSCs grown in neural induction medium (NI) containing recombinant human Wnt1 (10 or 100 ng/mL) for 7 days. PC; E10 otocyst cDNA, NC; no cDNA. (B) RT-PCR analysis for Dopaminergic (TH and Nurr1), GABAergic (Grik2 and Viaat) neuronal subtype and Glia (GFAP and Itgb4)) marker genes in MSCs grown in neural induction medium (NI) containing recombinant human Wnt1 (10, 100 and 400ng/mL) for 7 days. PC; E18 brain cDNA, NC; no cDNA. L27 is an internal control. Supplemental Figure 5 Expression of hematopoietic cell markers is not altered by Wnt1 in neurally-induced MSCs Flow cytometry profiles of CD34-FITC, CD117-APC and SCA1-FITC fluorescence in MSCs at neural induction day 7 without (NI) or with Wnt1 (NI+Wnt1). Supplemental Figure 6 Wnt1-induced up-regulation of CyclinD1 is suppressed by Dkk1 or sFRP2 in neurally-induced MSCs Western blot analysis for CyclinD1 in MSCs grown in neural induction medium with recombinant human Wnt1 and either Dkk1(100 ng/mL) or sFRP2 (100 ng/mL) for 7 days. β-actin is used as a loading control. Supplemental Figure 7 The extent of cell proliferation is not altered by Wnt1 in neurally-induced MSCs (A, B) BrdU immunofluorescence in MSCs grown in neural induction medium without (A) or with (B) Wnt1 for 7 days. (C) The percentage of BrdU-positive cells in neurally-induced MSCs grown in the presence or absence of Wnt1.|
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