Project description:Colon cancer patient-derived xenograft (PDX) models were processed to single cells and sorted by FACS (BD FACS Aria II) for ALDH activity (Aldefluor assay) and DAPI. ALDH Negative and ALDH Positive cells from each PDX model were collected and lysed in RLT buffer and processed for RNA using the RNeasy Mini Plus RNA extraction kit (Qiagen). Samples were processed using Illumina’s TrueSeq RNA protocol and sequenced on an Illumina HiSeq 2500 machine as 2x125nt paired-end reads. Reads were mapped to the human reference genome (assembly hg19) using the STAR aligner (version 2.4.2a). Total read counts per gene were computed using the program “featureCounts” (version 1.4.6-p2) in the “subread” package, with the gene annotation taken from Gencode (version 19).
Project description:Fluorescence-activated cell sorting (FACS) was used to separate colon tumor samples into two fractions of ALDH Negative and ALDH Positive cells using the Aldefluor Assay. RNA samples were derived from FAC-sorted 3D in-vitro models of patient-derived colon tumors. Samples from unfractionated in vitro models are also included.
Project description:The intent of this experiment was to determine the similarities and differences in expression signatures of normal breast cells that express markers associated with an epithelial-like (ALDH+) and mesenchymal-like (CD44+CD24-) stem cells in the normal human breast. Briefly, tissues were collected from women undergoing voluntary reduction mammoplasty. Tissues were dissociated using chemical and enzymatic methods to yield single cells. These cells were sorted by flow cytometry for aldehyde hydrogenase activity (ALDH+), CD44, and CD24, as well as viability, following depletion of hematopoeitc cells, endothelial cells, and fibroblasts.
Project description:Purpose: we aimed at identify and compare the transcriptional changes of ALDH- and ALDH+ DU145 xenografts upon radiotherapy treatment. Method: Xenografts were generated by injection of ALDH- and ALDH+ DU145 cells in male NMRI-Foxn1 nu/nu immune-deficient mice and subjected to fractionated irradiation to a final dose of 50 Gy. Tumors were excised and processed for total RNA extraction and RNAseq analysis.
Project description:To identify genes that are differentially expressed between ALDH-high and ALDH-low cells in endometrial cancer-derived spheroids, we performed whole genome microarray expression profiling as a discovery platform. Human endometrial cancer stem spheroid cells from three patients were sorted into ALDH-high and ALDH-low cells using the FACS Aria II Cell Sorter. Total RNA extracted from the cells were labeled with Cy3 and used for microarray analyses with Agilent Whole Human Genome Oligo Microarrays.
Project description:Transcriptional profiling of cancer stem cells (ALDH-high cells) comparing non-cancer stem cells (ALDH-low cells), sorted out using ALDEFLUOR assay. Goal was to identity cancer stem cell-specific genes. Two-condition experiment, sphere-cultured cells vs. adherent-cultured cells: 1 ALDH-high replicate and 1 ALDH-low replicate.
Project description:We used an Aldefluor assay system to study the signaling pathways that regulate the frequency and maintenance of ALDH-positive cell population in MCF-7 cells as a CSC model. Through microarray analysis, we investigated a possible receptor that increases the proportion of the ALDH-positive cell population in MCF-7 cells. MCF-7 were stained with Aldefluor and ALDH-positive cells were sorted using flow cytometry. Gene expression profiles of ALDH+ cells were compared with ALDH- cells.
