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Embryonic Stem Cells/Induced Pluripotent Stem Cells
Article first published online: 22 MAY 2013
Copyright © 2013 AlphaMed Press
Volume 31, Issue 6, pages 1086–1096, June 2013
How to Cite
Ogaki, S., Shiraki, N., Kume, K. and Kume, S. (2013), Wnt and Notch Signals Guide Embryonic Stem Cell Differentiation into the Intestinal Lineages. STEM CELLS, 31: 1086–1096. doi: 10.1002/stem.1344
Author contributions. S.O.: acquisition of data and analysis and interpretation of data; N.S.: study concept and design and acquisition, analysis, and interpretation of data; K.K.: critical revision of the manuscript and administrative and study supervision; S.K.: study concept and design, drafting and revision of the manuscript, approval of the final version of the manuscript, and obtained funding.
Disclosure of potential conflicts of interest is found at the end of this article.
first published online in STEM CELLS EXPRESS February 4, 2013.
- Issue published online: 22 MAY 2013
- Article first published online: 22 MAY 2013
- Accepted manuscript online: 4 FEB 2013 01:41AM EST
- Manuscript Accepted: 10 JAN 2013
- Manuscript Received: 14 AUG 2012
- Cell Fate Regulation Research and Education Unit. Grant Numbers: #21390280, #21790671
- Funding Program for Next Generation World-Leading Researchers
- Japan Society for the Promotion of Science
- Realization of Regenerative Medicine
- Ministry of Education, Culture, Sports, Science and Technology (MEXT) Japan
Additional Supporting Information may be found in the online version of this article.
|sc-12-0751_sm_SupplFigure1.pdf||239K||Supplementary Figure 1 Cdx2+ cells are proliferative cells EdU incorporation tests assayed for 3 hours (d12) revealed that 45.6±1.51% (n=3) of the ES cell-derived Cdx2+ (green) cells were EdU positive (red). DAPI (blue) indicate nuclei. Scale bars: 100 μm.|
|sc-12-0751_sm_SupplFigure2.tif||2027K||Supplementary Figure 2 MEF cells did not express Cdx2. (A) Immunocytochemical analysis of MEF treated with BIO and DAPT. RT-PCR analyses of intestinal markers on day 20. ES cells differentiated on MEF without BIO and DAPT. Scale bars: 100 μm. (B) Immunocytochemical analysis using specific isotype negative control IgGs. Scale bars: 100 μm.|
|sc-12-0751_sm_SupplFigure3.tif||1054K||Supplementary Figure 3 There were no undifferentiated ES on day 15 under feeder free condition. Immunocytochemical analysis of undifferentiated ES cells and ES cells-derived cells on day 15 are shown. Nanog+ cells were not observed on day 15. Scale bar: 100 μm.|
|sc-12-0751_sm_SupplFigure4.tif||136K||Supplementary Figure 4 FGF inhibitor SU5402 treatment increased initial induction of CDX2. (A) A schematic presentation of the experiment. khES3 cells were cultured on feeders, added with SU5402 from day 10 to 30, and were assayed for CDX2 transcript by real time PCR. (B) Initial induction of CDX2 was potentiated by SU5402, but CDX2 expression in control caught up later. **p < 0.01 versus control (white bars) by Student's t-test (N=3-5).|
|sc-12-0751_sm_SupplFigure5.tif||2100K||Supplementary Figure 5 FGF2 inhibited intestinal differentiation in Caco2 cells. (A) A schematic presentation of the experiment. Caco2 cells were cultured to confluency, then differentiated at the presence or absence of FGF2. (B) Bright field views of Caco2 cells under different FGF2 conditions. FGF2 inhibited dome-like morphology of mature intestinal endoderm. (C, D) Flow cytometeric analysis of Caco2 cells treated with graded concentrations of FGF2. Differentiation into CDX2high population was inhibited by 250 ng/ml FGF. (E) RT-PCR analysis of Caco2 cells treated with FGF2. FGF2 inhibited CDX2 expression in Caco2 cells. **p < 0.01 versus control (white bar) by Student's t-test (N=3). Scale bars: 100 μm.|
|sc-12-0751_sm_SupplTable1.pdf||76K||Supplementary Table 1 The primer sequences and number of cycles. The primer sequences, cycle numbers for semi-quantitative RT-PCR and real time PCR are shown.|
|sc-12-0751_sm_SupplTable2.pdf||87K||Supplementary Table 2 The growth factors tested for potentiating the induction into Cdx2-positive intestinal cells. The growth factors and suppliers used in the experiments are shown.|
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