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Tissue-Specific Stem Cells
Version of Record online: 22 OCT 2012
Copyright © 2012 AlphaMed Press
Volume 30, Issue 11, pages 2487–2497, November 2012
How to Cite
Paik, D. Y., Janzen, D. M., Schafenacker, A. M., Velasco, V. S., Shung, M. S., Cheng, D., Huang, J., Witte, O. N. and Memarzadeh, S. (2012), Stem-Like Epithelial Cells Are Concentrated in the Distal End of the Fallopian Tube: A Site for Injury and Serous Cancer Initiation. STEM CELLS, 30: 2487–2497. doi: 10.1002/stem.1207
Author contributions: D.Y.P. and D.M.J.: conception and design, collection and/or assembly of data, data analysis and interpretation, and manuscript writing; A.M.S., V.S.V., M.S.S., and D.C.: conception and design, collection and/or assembly of data, and data analysis and interpretation; J.H.: data analysis and interpretation; O.N.W.: manuscript writing; S.M.: conception and design, data analysis and interpretation, manuscript writing, 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 August 21, 2012.
- Issue online: 22 OCT 2012
- Version of Record online: 22 OCT 2012
- Accepted manuscript online: 21 AUG 2012 08:21AM EST
- Manuscript Accepted: 11 JUL 2012
- Manuscript Received: 5 MAY 2012
- Liz Tilberis Scholars Program
- Ovarian Cancer Research Fund, Inc.
- Gynecologic Cancer Foundation
- St. Louis Ovarian Cancer Awareness Research Grant
- Ovarian Cancer Circle
- Howard Hughes Medical Institute investigator
Additional Supporting Information may be found in the online version of this article.
|sc-12-0431_sm_SupplFigure1.tif||1040K||Supporting Information Figure S1. FTE growth is clonal and cells within large spheres can self-renew. (A) Representative field illustrating classification of large (>250 μm) and small (<250 μm) spheres. (B) The numbers of spheres generated was linear with respect to cells plated in vitro. (C) Fallopian tube epithelia were color marked with a lentivirus expressing the red fluorescent protein (RFP). Large vs. small RFP marked spheres were hand-picked and dissociated separately. Twenty-five hundred color labeled cells from each condition (large vs. small spheres) were combined with equal numbers of unlabeled carrier cells from spheres and plated in vitro. Numbers of RFP marked large vs. small spheres were quantified and compared. Only large spheres were capable of giving rise to large and small spheres. Small spheres only gave rise to small spheres but could not generate large spheres. (D) Cells in FTE spheres could be dissociated and re-grown for multiple passages.|
|sc-12-0431_sm_SupplFigure2.tif||713K||Supporting Information Figure S2. Gating strategy for the isolation of FTESC cells and fold enrichment of in vitro growth for FTESC+ vs. FTESC- vs. bulk FTE. Examples of cells isolated from the FTE and processed through a FACS sorter from postmenopausal (A) and pre-menopausal (B) specimens are shown. The gating strategy was the same for all specimens. Lineage positive single cells were removed by gating out PTPRC-PECAM1-GYPA- cells (Aa, Ba) while epithelial cells were selected by gating on the EPCAM+ population (Ab, Bb). The CD44+ITGA6hi gate was set using the fluorescence minus one (FMO) approach which entails staining with all antibodies except anti-CD44 (Ac, Bc). Based on this gating EPCAM+CD44+ITGA6hi FTESC cells were detected on fully stained samples (anti-CD44 antibody included) (Ad, Bd). (C) In post-menopausal specimens, FTESC was 16.5 fold (± 4.3x) enriched for cells with large sphere forming capacity compared to the FTESC- fraction and 4.8 fold (± 0.1x) compared to bulk FTE (a). In pre-menopausal specimens, FTESC was 16.8 fold (± 0.4x) enriched for cells with large sphere forming capacity compared to the FTESC- fraction and 5.7 fold (± 1.0x) compared to bulk FTE(b).|
|sc-12-0431_sm_SupplFigure3.pdf||261K||Supporting Information Figure S3. Vast majority of normal ovarian surface epithelia (OSE) do not express CD44. The expression of CD44 was examined in the normal ovarian surface epithelia of eight independent patients. The OSE was identified by expression of pan-keratin on the surface of the ovary. Predominance of ovarian surface epithelia did not express CD44. Scale bars equal 50 μm.|
|sc-12-0431_sm_SupplFigure4.pdf||205K||Supporting Information Figures S4. An expansion of CD44 and KRT5 positive cells was detected in tubal intraepithelial carcinomas (TIC). CD44 and KRT5 expression was examined in two independent patient samples with a diagnosis of TIC based on histology and TP53 expression. In both specimens, an expansion of cells expressing CD44 and KRT5 was observed in the abnormal epithelium. Scale bars equal 50 μm.|
|sc-12-0431_sm_SupplMovies1.wmv||223K||Supporting Information Video S1. FTE spheres contain beating cilia in the luminal space. Functional ciliated cells were detected in FTE spheres arising from single cells supporting multi-lineage differentiation of FTE. Field of view is 450 μm.|
|sc-12-0431_sm_SupplTable1.tif||10K||Supplementary Table 1|
|sc-12-0431_sm_SupplTable2.tif||23K||Supplementary Table 2|
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