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Profiling of normal human colonic epithelial cells according to their PTK7 surface abundance

ABSTRACT: Human mucosa was collected from two different individuals undergoing colectomy. After treatment with EDTA, colonic crypts were isolated and further dis-aggregated using Dispase. Next, cells were stained using antibodies directed against the extracellular domain of EpCAM, PTK7, CD11, CD31, and CD45. Cells were analyzed and sorted via flow cytometry (BD Aria). After excluding non-epithelial cells (Epcam. CD11, CD31, and CD45 negative), the EpCAM positive fraction was divided into fractions expressing either high, medium, low, or negative levels of PTK7. Sorted cells were transferred to Trizol for RNA extraction and RNA was purified using the Qiagen RNA Mini Kit.

ORGANISM(S): Homo sapiens

PROVIDER: GSE68340 | GEO | 2016/01/12

SECONDARY ACCESSION(S): PRJNA282480

REPOSITORIES: GEO

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Profiling of normal human colonic epithelial cells according to their PTK7 surface abundance

Project description:Human mucosa was collected from two different individuals undergoing colectomy. After treatment with EDTA, colonic crypts were isolated and further dis-aggregated using Dispase. Next, cells were stained using antibodies directed against the extracellular domain of EpCAM, PTK7, CD11, CD31, and CD45. Cells were analyzed and sorted via flow cytometry (BD Aria). After excluding non-epithelial cells (Epcam. CD11, CD31, and CD45 negative), the EpCAM positive fraction was divided into fractions expressing either high, medium, low, or negative levels of PTK7. Sorted cells were transferred to Trizol for RNA extraction and RNA was purified using the Qiagen RNA Mini Kit. Colonic crypts were isolated from normal human mucosa derived from individuals undergoing colectomy. Single cell fractions from these crypts were sorted and isolated RNA processed and hybridized to Affymetrix PrimeView Arrays

2016-01-12 | E-GEOD-68340 | biostudies-arrayexpress

Transcription as a function of tissue environment

Project description:We are interested in whether fibroblasts (CD45-CD31-Epcam-PDPN+PDGFRA+) cells exhibit unique transcriptional profiles. To this end, we digested tissues and FACS sorted CD45-CD31-Epcam-PDPN+PDGFRA+ fibroblasts CD45-CD31-Epcam-PDPN+PDGFRA- mesothelial cells and compared the transcriptional profiles of these cells. PRJ0014647

2019-07-16 | GSE130813 | GEO

Signature of lung epithelial and mesenchymal populations from control, irradiated and neutrophil-depleted irradiated mice

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RNA-seq from HNSCC and melanoma populations

Project description:Populations were sorted from tumor speciments via flow cytometry using EPCAM, FAP, CD45, and CD31 surface markers

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Sentinel p16INK4a+ cells in the basement membrane form a reparative niche in the lung [RNA-Seq]

Project description:We collected freshly sorted mesenchyme (live CD45-Epcam-CD31-PDGFRa+GFP- or GFP+) from dissociated lung tissue of INKBRITE animals to characterize the transcriptome difference between p16INK4a+ vs p16INK4a- cells at homeostasis and during injury. We used fluorescence activated cell sorting (FACS) to exclude immune (CD45+), epithelial (Epcam+), and endothelial (CD31+) cells and positively select for PDGFRa+ mesenchyme. Cells were sequenced at a depth of average of 45 million reads/sample. The results revealed an increase of senescence associated secretory phenotype (SASP) signature in p16INK4+ cells that becomes refined after injury. We found the expression of an epithelial growth factor epiregulin (ereg) increased in p16INK4a+ cells after injury suggesting a possible role of p16INK4a+ senescence during regeneration of lung epithelium.

2022-11-02 | GSE140654 | GEO

Expression profiles of cell populations purified from human CRC (4 ways)

Project description:The survival of isolated metastatic cells and expansion into macroscopic tumour has been recognized as a limiting step for metastasis formation in several cancer types yet the determinants of this process remain largely uncharacterized. In colorectal cancer (CRC), we identify a transcriptional programme in tumour-associated stromal cells, which is intimately linked to a high risk of developing recurrent disease after therapy. A large proportion of CRCs display mutational inactivation of the TGF-beta pathway but paradoxically they are characterized by high TGF-beta production. In these tumours, TGF-beta instructs a transcriptional programme in stromal cells, which confers a high risk of developing metastatic disease. We purified by FACS CD31(+), CD45(+), FAP(+) and Epcam(+) populations from fresh CRC samples and assessed their gene expression profiles Freshly obtained tumours from 6 CRC patients treated at Hospital del Mar or Hospital Clinic (Barcelona, Spain) were minced and incubated with Collagenase IV (100 U mL-1). Enzymatic reaction was stopped by adding 10% FBS, single cells were collected by sequential filtering and resuspended in ammonium chloride (0.15M) to lyse erythrocytes. Cells were stained with FAP unconjugated rabbit antibody. After two washes with HBSS, cells were stained with an APC conjugated anti-rabbit antibody; anti-hEPCAM/TROP1-FITC, anti-CD31-PE and anti-CD45-PE-Cy7 conjugated antibody. Dead cells were labelled with Propidium Iodide. Fluorescence Activated Cell Sorting (FACS) was used to separate 2000 cells from each population; [CD45(+), EPCAM(-), CD31(-), FAP(-)], [CD45(-) EPCAM(+), CD31(-), FAP(-)], [CD45(-), EPCAM(-), CD31(+),FAP(-)] and [CD45(-), EPCAM(-), CD31(-), FAP(+)].

2013-01-17 | E-GEOD-39396 | biostudies-arrayexpress

RNA-seq profiling of gene expression in Ezh2-deficient and control murine lung epithelial and stromal cell populations at day E16.5

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2015-05-11 | GSE57391 | GEO

Expression data of PTK7+ and PTK7- cells from differentiated pluripotent stem cells (PSC), and expression data from undifferentiated PSC and definitive endoderm

2012-11-07 | GSE42073 | GEO

Expression data of PTK7+ and PTK7- cells from differentiated pluripotent stem cells (PSC), and expression data from undifferentiated PSC and definitive endoderm

Project description:As PSCs differentiate, they display characteristics of EMT as well as PTK7 expression. We sought to discover novel markers and regulators of this process by comparing purified PTK7+ and PTK7- populations We used microarrays to detail the global program of gene expression underlying this EMT process and identified distinct classes of up-regulated genes during this process. PTK7+ and PTK7- populations were purified by FACS and subjected to lysis and RNA extraction. Undifferentiated PSCs and definitive endoderm were washed with PBS, then subjected to lysis and RNA extraction

2012-11-07 | E-GEOD-42073 | biostudies-arrayexpress

The CD90+CD39+ and CD90+CD39- fibroblast subtypes in human hypertrophic scar and normal skin

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2021-07-05 | GSE158766 | GEO

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