Project description:To prove that hypoxia generates Cancer stem cells (CSCs) from non-CSCs, we needed to separate out already existing tumorigenic cells from the cell population. To identify CSCs, we used ES cell-specific cell surface antigens (SSEA-1, -3, -4, and Tra1-81) on the assumption that the immature cell fraction would be rich in CSCs. We analyzed the expression levels of such embryonic antigens and stemness genes in cells exposed to hypoxia. iPS (253G1), SSEA-1 positive-N417 3 of SSEA-1/3 double negative-N417, each condition is represented by 3 biological replicates
Project description:A subset of cells in cellular populations may be preferentially reprogrammed into pluripotent stem cells due to their innate gene expression characteristics. SSEA-1+ sorted cells were shown to be more amenable to reprogramming than other cells in porcine fetal fibroblast cell lines. This microarray data reveals which genes are unique to the SSEA1+ sorted cells and helps to give them an identity. Porcine fetal fibroblasts were subjected to Magnetic Activated Cell Sorting using anti-SSEA-1. Five biological replicates were performed resulting in 5 SSEA-1 positive fractions and 5 SSEA-1 negative fractions which were analyzed.
Project description:Comparison of gene expression profile of different types of cells: hESCs (H1), human adult dermal fibroblasts, cells from adult human ovarian cell culture, FACS sorted SSEA-4 positive cells from adult human ovarian culture and cells from human ovarian cancer cell culture.
Project description:Direct comparison of FACS sorted EGFP positive brain macrophages vs pooled Total Brain extract Keywords: Direct pairwise comparison
Project description:Direct comparison of FACS sorted EGFP positive renal macrophages vs pooled Total Kidney extract Keywords: Direct pairwise comparison
Project description:Transcriptional profiling of ALDH1 high cancer stem like cells of ovarian cancer cell line, sorted out using ALDEFLUOR assay. Two-condition experiment, ALDEFLUOR positive vs. negative cells. Biological replicates: 1 ALDEFLUOR positive replicate, 1 negative replicate.
Project description:MSCs comprise several percent of pluripotent-like cells named Multilineage-differentiating stress enduring (Muse) cells that express pluripotent markers at moderate levels, are collectable as cells positive for the pluripotent surface marker SSEA-3, are able to differentiate into triploblastic lineage cells, and self-renew at the single cell level (Kuroda et al, PNAS, 2010; Wakao et al, PNAS, 2011). Importantly, MSCs also comprise cells other than SSEA-3(+)-Muse cells, namely multipotent SSEA-3(-)-non-Muse MSCs that correspond to ~98% of the total MSC population. These SSEA-3(-)-non-Muse MSCs exhibit the same properties as conventional MSCs, although the non-Muse MSCs are multipotent, they are not pluripotent. In the present study, to clarify the key molecules that characterize pluripotent-like vs multipotent somatic stem cells, we separated human MSCs into SSEA-3(+)-Muse cells and SSEA-3(-)-non Muse MSCs, and analyzed both populations by scRNA-seq. SSEA-3(+) Muse cells and SSEA-3(-) non-Muse MSCs were sorted from human BM-MSCs. Sequencing libraries were prepared using Chromium single cell 3' Kit v3 (10x Genomics, Pleasanton, CA, USA) and sequenced on HiSeq2500 (Illumina, San Diego, CA, USA). Transcripts were mapped with CellRanger pipeline v3 (10x Genomics). Library construction, sequencing, and initial analysis were performed by GENEWIZ (South Plainfield, NJ, USA).
Project description:We conducted bulk RNA sequencing on cells sorted by fluorescence from micropatterned colonies treated with FGF8 and different concentrations of WNT3a, using a SOX2::mCitrine cell line to distinguish between SOX2 positive and negative cells. The sequencing revealed that SOX2 negative cells exhibited a definitive endoderm identity, while SOX2 positive cells showed epiblast identity.
