Additional Supporting Information may be found in the online version of this article.

sc-12-0431_sm_SupplFigure1.tif1040KSupporting 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.tif713KSupporting 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.pdf261KSupporting 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.pdf205KSupporting 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.wmv223KSupporting 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.tif10KSupplementary Table 1
sc-12-0431_sm_SupplTable2.tif23KSupplementary Table 2

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