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Cancer Stem Cells
Article first published online: 22 MAR 2012
Copyright © 2012 AlphaMed Press
Volume 30, Issue 4, pages 591–598, April 2012
How to Cite
Xin, H.-W., Hari, D. M., Mullinax, J. E., Ambe, C. M., Koizumi, T., Ray, S., Anderson, A. J., Wiegand, G. W., Garfield, S. H., Thorgeirsson, S. S. and Avital, I. (2012), Tumor-Initiating Label-Retaining Cancer Cells in Human Gastrointestinal Cancers Undergo Asymmetric Cell Division. STEM CELLS, 30: 591–598. doi: 10.1002/stem.1061
Author contributions: H.-W.X: concept and design, collection and assembly of data, data analysis and interpretation, and manuscript writing; D.M.H., J.E.M., C.M.A., A.J.A., G.W.W., and S.H.G.: collection and assembly of data; T.K. and S.R.: manuscript writing; S.S.T.: data analysis and interpretation and manuscript writing; I.A: concept and design, data analysis and interpretation, manuscript writing, and final approval of manuscript. H.-W.X. and D.M.H. 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 February 13, 2012.
- Issue published online: 22 MAR 2012
- Article first published online: 22 MAR 2012
- Accepted manuscript online: 13 FEB 2012 04:43PM EST
- Manuscript Accepted: 29 JAN 2012
- Manuscript Received: 16 DEC 2011
- NIH/National Cancer Institute
Additional Supporting Information may be found in the online version of this article.
|STEM_1061_sm_SuppFig1.tif||1098K||Figure S1. A subpopulation of Label-Retaining-Cancer-Cells (LRCC) is not quiescent and undergoes active cell-division. (A) Experimental procedure for the isolation of live LRCC and non-LRCC. (B) A FACS diagram shows Cy5+ LRCC (green) were successfully sorted out after 15 cell generations of growth in log phase post initial Cy5-dUTP labeling and sorting. (C-D) FACS analysis of Ki67+ cells show that a subpopulation of LRCC isolated from human cancer cell line and fresh primary human cancer cells are proliferating with 89.4% ± 3.3 of Ki67+ cells comparing to 79.2% ± 5.2 for non-LRCC (p=0.20). (E-F) FACS analysis of pHH3+ cells show that a subpopulation of LRCC isolated from human cancer cell line and fresh primary human cancer cells are actively dividing with 13.5% ± 2.5 of pHH3+ cells comparing to 6.5% ± 1.6 for non-LRCC (p=0.078). (G-I) Cell cycle analysis show that a subpopulation of LRCC isolated from human cancer cell line and fresh primary human cancer cells undergo active cell division with 16.9% ± 3.4 of G2/M phase cells comparing to 11.6% ± 2.7 for non-LRCC (p=0.28).|
|STEM_1061_sm_SuppFig2.tif||2471K||Figure S2. Cy5-dUTP labels DNA within nuclei in the time lapse confocal imaging experiments for real time detection of ACD-NRCC of live LRCC. (A) Experimental procedure of real time detection of live LRCC undergoing ACD-NRCC by DNA labeling with fluorescent Cy5-dUTP and time lapse confocal microscopy imaging. (B-C) Confocal images show that Cy5-dUTP co-localizes with DAPI staining supporting Cy5- dUTP labeling of DNA within nuclei. (D-E) Three dimensional confocal images re-constructed from serial single confocal images show that Cy5-dUTP labels DNA within nuclei.|
|STEM_1061_sm_SuppTab1.pdf||21K||Table S1. Determination of cell doubling times. To effectively detect asymmetric cell division, we determined the growth curves and doubling times experimentally for all cell lines and the fresh tested tumor cells. Growth curve's correlation value R2≥0.9 was considered adequate for computations of doubling times.|
|STEM_1061_sm_SuppVideo1.avi||5624K||Movie S1. A time-laspe confocal microscopy movie showing LRCC undergoing ACD-NRCC in live cells and in real-time.|
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