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Brief Report: Embryonic Stem Cells/Induced Pluripotent Stem Cells
Version of Record online: 16 NOV 2011
Copyright © 2011 AlphaMed Press
Volume 29, Issue 12, pages 2094–2099, December 2011
How to Cite
Son, Y. S., Seong, R. H., Ryu, C. J., Cho, Y. S., Bae, K.-H., Chung, S. J., Lee, B., Min, J.-K. and Hong, H. J. (2011), Brief Report: L1 Cell Adhesion Molecule, a Novel Surface Molecule of Human Embryonic Stem cells, Is Essential for Self-Renewal and Pluripotency. STEM CELLS, 29: 2094–2099. doi: 10.1002/stem.754
Author contributions: Y.S.S.: conception and design, collection and assembly of data, and data analysis and interpretation; R.H.S.: administrative support and data analysis and interpretation; C.J.R.: conception and design; Y.S.C. and K.-H.B.: administrative support, data analysis, and financial support; S.J.C.: collection and assembly of data and financial support; B.-H.L.: administrative support and financial support; J.-K.M.: conception and design, collection and assembly of data, data analysis and interpretation, financial support, manuscript writing, and final approval of manuscript; H.J.H.: conception and design, data analysis and interpretation, financial support, and final approval of manuscript.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLSEXPRESS September 28, 2011.
- Issue online: 16 NOV 2011
- Version of Record online: 16 NOV 2011
- Accepted manuscript online: 28 SEP 2011 03:02PM EST
- Manuscript Accepted: 16 SEP 2011
- Manuscript Received: 19 MAY 2011
- Korea Research Institute of Bioscience & Biotechnology
- Korea Research Council of Fundamental Science and Technology
- Stem Cell Research Program
- Korea Science and Engineering Foundation. Grant Numbers: M1AN41-2006-04440, SC12023
- Stem Cell Research Center of the 21st Century Frontier Research Program
- Ministry of Science and Technology of Korea
Additional Supporting Information may be found in the online version of this article.
|STEM_754_sm_supplFigure1.pdf||643K||Figure S1. Identification of MAb 4-63 antigen. (A) Extracts of biotinylated H9 hESCs were immunoprecipitated with 4-63 or no antibody (bead), followed by Western blot analysis using streptavidin-HRP. 4-63 in the non-IP section indicates that only 4-63 antibody was loaded. (B) Sequence analysis of L1CAM expressed in hESCs. The MS/MS spectrum of L1CAM obtained after trypsin digestion was analyzed by Q-TOF mass spectrometry. The precursor ion shown in the figure is m/z 673.93; resultant peaks were searched against the NCBInr database. Four tryptic peptides that matched the L1CAM isoform 2 are underlined in the sequence presented in B. The hESC-expressed L1CAM was the non-neuronal isoform because RSLE was not present in the peptide (N-DETFGEYSDNEEK-C) sequenced from the precursor ion of m/z 781.88. To confirm this result, we cloned the full-length cDNA encoding L1CAM from H9 cells and determined its entire nucleotide sequence. The L1CAM sequence lacks the YEGHH and RSLE peptides found in neural L1CAM.|
|STEM_754_sm_supplFigure2.tif||2492K||Figure S2. Specificity of L1CAM expression in undifferentiated hESCs and mESCs. (A) H9 cells and day 12-derived EBs were incubated with anti-SSEA-3, anti-SSEA-4, or L1CAM, followed by FITC-conjugated anti-rat IgM or anti-mouse IgG. Fluorescence is compared with control in each panel. (B-C) Total mRNAs were isolated from the hESCs (B) or msESC (C) cultured for 6 or 12 days, or the EBs derived at indicated days. The expression level of transcription factors essential for the maintenance of an undifferentiated state in ESCs (Nanog, Oct4, and Sox2) and three germ layer markers (ectoderm, Pax6; mesoderm, CD34; and endoderm, AFP) was assessed by RT-PCR.|
|STEM_754_sm_supplFigure3.tif||1922K||Figure S3. AP staining in H9-derived stable cell lines. (A) AP staining of H9-derived stable cell lines. (B) The graph shows the data of AP-positive cells. L1CAM depletion in hESCs significantly reduced AP staining. In contrast, L1CAM-overexpressing cells strongly expressed AP. Three independent experiments were performed in duplicate. Data are presented as means ± SD (**p < 0.01).|
|STEM_754_sm_supplTable1.pdf||113K||Supplementary Table 1.|
|STEM_754_sm_supplTable2.pdf||76K||Supplementary Table 2.|
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