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A Comparison of Whole Genome Gene Expression Profiles of 0p-EpSCs and 15p-EpSCs

ABSTRACT: To investigate successful maintenance of skin epithelial stem cells (EpSCs) in vitro using Wnt-3a, we prepared CD49f+CD34+ cells, an EpSC-rich population from adult mouse skin and cultured for 10 days in the presence of Wnt-3a. Sorted CD49f+CD34+ cells proliferated markedly in the presence of Wnt-3a. Those cells were then subjected to a second 10-day culture with Wnt-3a and sorted, with the same procedures repeated until fifteen 10-day cultures were performed. A dye-swapped experiment was performed by hybridizing complimentary RNA (cRNA) labeled with either Cyanine (Cy) -3 or Cy-5 onto 4X44K ver.2 Agilent Whole Mouse Genome Oligo Microarray (G4846A).

ORGANISM(S): Mus musculus

SUBMITTER: Daisuke Okuzaki

PROVIDER: E-GEOD-60503 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Partial maintenance and long-term expansion of murine skin epithelial stem cells by Wnt-3a in vitro.

Ouji Yukiteru Y Ishizaka Shigeaki S Nakamura-Uchiyama Fukumi F Okuzaki Daisuke D Yoshikawa Masahide M

The Journal of investigative dermatology 20141201 6

CD49f(+)CD34(+) cells, a skin epithelial stem cell (EpSC)-rich population, were prepared from adult mouse skin and cultured in the presence of Wnt-3a without feeder cells. CD34 expression was retained in about 10% of the cells, which had proliferated about 1,000-fold by day 10, although completely lost by day 14. CD49f(+)CD34(+) cells sorted on day 10 retained canonical Wnt-responsiveness, proliferated markedly in the presence of Wnt-3a, maintained undifferentiated epithelial cell marker express ...[more]

PMID: 25437427

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RNA Sequencing of LT-HSCs and ST-HSCs from healthy adult human Bone Marrow

Project description:The transcriptome of LT-HSC (CD34+CD38-CD45RA+CD90+CD49f+) and ST-HSC (CD34+CD38-CD45RA+CD90-CD49f-) from healthy adult human Bone Marrow Cells were assessed by RNA-seq.

2022-04-06 | E-MTAB-9892 | biostudies-arrayexpress

Human FZD6 phosphorylation LC-MS/MS

Project description:Frizzleds (FZD) function as receptors for secreted lipoglycoproteins of the Wingless/Int-1 (WNT) family, initiating an important signal transduction network in multicellular organisms. FZDs are classified as G protein-coupled receptors (GPCRs), which control a wide variety of physiological functions. GPCRs are well known to be regulated by phosphorylation, which can lead to specification of downstream signaling or receptor desensitization. The role and underlying mechanisms of phosphorylation of FZDs remains largely unexplored. Here, we investigated the phosphorylation of human FZD6. In order to assess FZD6 phosphorylation experimentally, we employed mass spectrometry. HEK293 cells expressing FZD6-GFP were treated with either control, WNT-3A or WNT-5A containing conditioned medium for 30 min and processed for LC-MS/MS analysis. Six phosphorylated serine residues were detected in the C-terminus of FZD6 − S592, S620, S629, S648, S653 and S656.

2018-10-18 | PXD011173 | Pride

Single Cell RNA Sequencing of human bone marrow LT-HSCs.

Project description:10x single cell sequencing of LT-HSCs (CD34+CD38-CD90+CD45RA-CD49f+) from human bone marrow.

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Global gene expression analysis of human dermal fibroblasts treated with Wnt 3a compared to control cells

Project description:human dermal fibroblasts were treated with 100 ng/ml recombinant mouse Wnt 3a for 8 h and 24 h and compared to untreated controls at 8 and 24 h. RNA was isolated and microarray analysis was performed with Illumina Human Sentrix-6 BeadChip arrays

2014-11-26 | E-MTAB-837 | biostudies-arrayexpress

A Comparison of Whole Genome Gene Expression Profiles of 0p-EpSCs and 15p-EpSCs

Project description:To investigate successful maintenance of skin epithelial stem cells (EpSCs) in vitro using Wnt-3a, we prepared CD49f+CD34+ cells, an EpSC-rich population from adult mouse skin and cultured for 10 days in the presence of Wnt-3a. Sorted CD49f+CD34+ cells proliferated markedly in the presence of Wnt-3a. Those cells were then subjected to a second 10-day culture with Wnt-3a and sorted, with the same procedures repeated until fifteen 10-day cultures were performed.

2015-07-07 | GSE60503 | GEO

The miRNA profiling of goat CD49f-positive and negative testicular cells

Project description:Purpose: miRNAs, a member of the small RNA, play critical roles in the mammalian spermatogenesis. Spermatogonia was the foundation of spermatogenesis and valuable for the study of spermatogenesis. However, it is still not clear that the expression profiling of the miRNAs in spermatogonia of dairy goat. Methods: The CD49f was one of the surface markers for spermatogonia enrichment by MACS. Therefore, we used CD49f microbeads antibody to purify CD49f-positive and negative cells of dairy goat testicular cells by MACS (Magnetic Activated Cell Sorting), and then in-depth analyzed the miRNA expression in these cells using Illumina sequencing technology. Results: The results of miRNAs expression profiling in purified CD49f-positive and negative testicular cells showed that 933 were miRNAs upregulated in CD49f-positive cells and 916 were miRNAs upregulated in CD49f-negative cells with a 2-fold increase, respectively; some spermatogonial stem cells(SSCs) specific miRNAs and marker genes in testis had a higher level expression in CD49f-positive testicular cells, such as miR-221, miR-23a, miR-29b, miR-24, miR-29a, miR-199b, miR-199a, miR-27a, miR-21. Conclusions: our comparative miRNAome data provided some useful miRNAs profiling data of dairy goat spermatogonia cells and suggested CD49f could be used to enrich dairy goat spermatogonia-like cells, including SSCs. miRNA profiles of goat CD49f-positive and negative testicular cells were generated by deep sequencing, in triplicate, using Illumina GAIIx

2014-04-25 | E-GEOD-57041 | biostudies-arrayexpress

Developmental changes in the in vitro activated regenerative activity of primitive mammary epithelial cells

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2013-08-13 | E-GEOD-46357 | biostudies-arrayexpress

Progesterone and estrogen receptors segregate into different cell subpopulations in the normal human breast

Project description:Gene expression profiles of normal human mammary epithelial cell subsets enriched for luminal or myoepithelial characteristics on the basis of EpCAM and CD49f expression. Transcription profiles were compared between luminal- and myoepithelial-enriched human MEC subsets following 3D culture from 4 individuals. Gene expression profiles of normal human mammary epithelial cell subsets enriched for mature, luminal progenitor or bipotent progenitor-enriched characteristics on the bais of CD49f, MUC1, CD10, CD133 and Thy1 expression Transcription profiles were compared between unsorted, mature, luminal progenitor-enriched and bipotent progenitor-enriched human MEC subsets following 3D culture from 2 individuals. Four replicates of luminal-enriched human MECs and 3 replicates of myoepithelial-enriched MECs Transcription profiles of two replicates of unsorted, mature, luminal progenitor-enriched and bipotent progenitor-enriched human MEC subsets were analysed following 3D culture

2012-08-17 | E-GEOD-31429 | biostudies-arrayexpress

Foxc1 reinforces quiescence in hair follicle stem cells

Project description:Quiescent stem cells are periodically activated to maintain tissue homeostasis or occasionally called into action upon injury. Molecular mechanisms that constitutively maintain stem cell identity or promote stem cell proliferation and differentiation upon activation have been extensively studied. However, it is unclear how quiescent stem cells maintain identity and reinforce quiescence when they transition from quiescence to activation. Here we show mouse hair follicle stem cell compartment induces a transcription factor, Foxc1, when activated. Importantly, deletion of Foxc1 in the activated but not quiescent stem cells compromises stem cell identity, fails to re-establish quiescence and subsequently drives premature stem cell activation.These findings uncover a dynamic, cell-intrinsic mechanism employed by hair follicle stem cells to reinforce stemness in response to activation. Poly(A)-enriched transcriptome RNA-seq on HFSCs isolated in WT and K14Cre cKO mice at anagen and early telogen stage of hair cycle.

2016-02-05 | E-GEOD-67404 | biostudies-arrayexpress

Analysis of Gsk-3-Deficient Embryonic Stem Cells Reveals Large-Scale Alterations in Gene Expression

Project description:The two vertebrate Gsk-3 isoforms, Gsk-3a and Gsk-3b, are encoded by distinct genetic loci and exhibit mostly redundant function in murine embryonic stem cells (ESCs). Here we report that deletion of both Gsk-3a and Gsk-3b in mouse ESCs results in significant changes in gene expression. In contrast, deletion of either Gsk-3a or Gsk-3b individually had little effect on gene expression. These data support the notion that Gsk-3 isoforms are functionally redundant in embryonic stem cells. In addition, we did not find the expected upregulation of known Wnt target genes. Our data suggests that Gsk-3-meidated regulation of gene expression in embryonic stem cells is complex, and likely involves affects on numerous signaling pathways. The study was designed to examine the changes in gene expression between wild-type, Gsk-3a-/-, Gsk-3b-/-, and Gsk-3a-/-;Gsk-3b-/- mouse embryonic stem cells.

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