Translational and Clinical Research
Article first published online: 26 OCT 2010
Copyright © 2010 AlphaMed Press
Volume 28, Issue 10, pages 1893–1904, October 2010
How to Cite
Swistowski, A., Peng, J., Liu, Q., Mali, P., Rao, M. S., Cheng, L. and Zeng, X. (2010), Efficient Generation of Functional Dopaminergic Neurons from Human Induced Pluripotent Stem Cells Under Defined Conditions. STEM CELLS, 28: 1893–1904. doi: 10.1002/stem.499
Author contributions: A.S. and J.P.: collection and/or assembly of data, data analysis and interpretation; Q.L.: data analysis and interpretation; P.M.: provision of study material; M.S.R.: data analysis and interpretation, manuscript writing; L.C.: provision of study material, final approval of manuscript; X.Z.: conception and design, data analysis and interpretation, manuscript writing, final approval of manuscript. A.S. and J.P. contributed equally to this article.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLS EXPRESS August 16, 2010.
- Issue published online: 26 OCT 2010
- Article first published online: 26 OCT 2010
- Accepted manuscript online: 16 AUG 2010 12:00AM EST
- Manuscript Accepted: 4 AUG 2010
- Manuscript Received: 8 JUL 2010
Additional Supporting Information may be found in the online version of this article.
|STEM_497_sm_SuppFig1.tif||622K||Supporting Information Figure 1. β-catenin-/- ESC are defective for endoderm/mesoderm differentiation. (A) qPCR expression analysis of endoderm markers (Sox17, and Gata6), mesoderm markers (Brachyury, and Eomes), and ectoderm marker (Nestin) upon embryoid body differentiation in wt and β-catenin-/- ESC.|
|STEM_497_sm_SuppFig2.tif||1033K||Supporting Information Figure 2. β-catenin-/- ESC exhibit decreased transcript levels of the pluripotency factors Oct4 and Nanog upon RA differentiation. (A) qRT-PCR analysis of Oct4, Nanog, Lrh-1, and β-catenin expression in wt and β-catenin-/- ESC upon RA-mediated differentiation. *, P < 0.05|
|STEM_497_sm_SuppFig3.tif||827K||Supporting Information Figure 3. CHIR99021 induces Lrh-1 in a β-catenin-dependent manner. (A) qPCR analysis of Lrh-1 Expression in wt and β-catenin-/- ESC upon 12 hours treatment with CHIR99021 (10 nM-5 μM) in the absence of LIF.|
|STEM_497_sm_SuppFig4.tif||1055K||Supporting Information Figure 4. Lrh-1-/- ESC exhibit decreased transcript levels of the pluripotency factors Oct4 and Nanog upon RA differentiation. (A) qRT-PCR analysis of Oct4, Nanog, Lrh-1, and β-catenin expression in wt and Lrh-1-/- ESC upon RA-mediated differentiation. *, P < 0.05|
|STEM_497_sm_SuppFig5.tif||616K||Supporting Information Figure 5. The induction of endoderm/mesoderm/ectoderm factors upon embryoid body differentiation is intact in Lrh-1-/- ESC. (A) qPCR expression analysis of endoderm markers (Sox17, and Gata6), mesoderm markers (Brachyury, and Eomes), and ectoderm marker (Nestin) upon embryoid body differentiation in wt and Lrh-1-/- ESC.|
|STEM_497_sm_SuppFig6.tif||909K||Supporting Information Figure 6. BIO is unable to maintain Oct4 expression in long-term culture. (A) Wild-type mES cells were cultured in the absence of LIF with media supplemented with BIO or DMSO vehicle for 7 days. Oct4 expression was examined by Western blot analysis and β-actin was probed as a loading control.|
|STEM_497_sm_SuppFig7.tif||1362K||Supporting Information Figure 7. Generation of β-catenin-/-(mycLrh-1) stably expressed ESC. (A) Western blot analysis of isolated clones. Heterologous recombinant colonies were cultured in the presence of LIF and G418. Transgene expression was confirmed by blotting for the myc-epitope alongside Lrh-1. All clones are absent for β-catenin, and β-actin served as a loading control.|
|STEM_497_sm_SuppTables1-3.doc||74K||Supporting Information Tables 1 to 3.|
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