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Direct comparison of HES2 ESC overexpressing CD30 variant with HES2 overexpressing GFP

ABSTRACT: HES2 ESC were infected with a lentivirus construct containing either CD30 (TNFRSF8) extracellular deficient variant or GFP, using the ViraPower™ Promoterless Lentiviral Gateway® Kits (Invitrogen). HES2 cells were recovered and grown for 19 passages. HES2 were commonly grown on Mouse Embryonic Feeder cells in the presence of KSOR media. Before collection of RNA, ES cells were FACs sorted with antibodies to GCMT-2 and CD9 to separate ES cells from differentiated cells and mouse feeders. The experiment was repeated in triplicate. The final aim was to deduce the effect of overexpressing the variant form of CD30 on gene expression in HES2 cells.

ORGANISM(S): Homo sapiens

PROVIDER: GSE13261 | GEO | 2009/10/10

SECONDARY ACCESSION(S): PRJNA109549

REPOSITORIES: GEO

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Direct comparison of HES2 ESC overexpressing CD30 variant with HES2 overexpressing GFP

Project description:HES2 ESC were infected with a lentivirus construct containing either CD30 (TNFRSF8) extracellular deficient variant or GFP, using the ViraPowerâ?¢ Promoterless Lentiviral GatewayÂ® Kits (Invitrogen). HES2 cells were recovered and grown for 19 passages. HES2 were commonly grown on Mouse Embryonic Feeder cells in the presence of KSOR media. Before collection of RNA, ES cells were FACs sorted with antibodies to GCMT-2 and CD9 to separate ES cells from differentiated cells and mouse feeders. The experiment was repeated in triplicate. The final aim was to deduce the effect of overexpressing the variant form of CD30 on gene expression in HES2 cells. Three consecutive passages of HES2 cells over-expressing either CD30V or GFP were grown and subject to FACs sorting, collection and microarray. A total of 6 samples, (3 replicates of these 2 populations) were used in the experiment.

2010-04-08 | E-GEOD-13261 | biostudies-arrayexpress

Gene expression profiling of granuloma T cells in M. tuberculosis infected rhesus macaques reveals a critical role for CD30 [Macaca mulatta]

Project description:Control of Mycobacterium tuberculosis infection requires generation of T cells that migrate to granulomas, complex immune structures surrounding sites of bacterial replication. Here we compared the gene expression profiles of T cells in pulmonary granulomas, bronchoalveolar lavage and blood of Mtb-infected rhesus macaques to identify granuloma-enriched T cell genes. TNFRSF8/CD30 was among the top genes that was upregulated in both CD4 and CD8 granuloma T cells and independent of bacterial loads. Transcriptomic profiling of lung T cells from Mtb-infected mixed bone marrow chimeric mice showed that CD30 directly promotes CD4 T cell differentiation and effector molecule expression. Moreover, in mice CD30 expression on CD4 T cells is required for survival of Mtb infection. These results show the CD30 co-stimulatory axis is highly upregulated on granuloma T cells and is critical for the generation of protective T cell responses against Mtb infection.

2023-03-22 | GSE227653 | GEO

Gene expression profiling of granuloma T cells in M. tuberculosis infected rhesus macaques reveals a critical role for CD30 [Mus musculus]

2023-03-24 | GSE228113 | GEO

HES2 embryonic stem cells (ESCs) grown in standard conditions and FACs sorted for GCTM-2 and CD9

Project description:HES2 ESCs were grown in standard ES culture conditions. After 1 week, these cells were FACs sorted for the presence of GCTM-2 and CD9. Keywords: Sorted cell comparison

2009-10-01 | GSE13201 | GEO

Homo sapiens

Project description:Direct comparison of HES2 ESC overexpressing CD30 variant with HES2 overexpressing GFP

| PRJNA109549 | ENA

HES2 embryonic stem cells (ESCs) grown in standard conditions and FACs sorted for GCTM-2 and CD9

Project description:HES2 ESCs were grown in standard ES culture conditions. After 1 week, these cells were FACs sorted for the presence of GCTM-2 and CD9. Keywords: Sorted cell comparison Three consecutive passages of HES2 cells were grown and subjected to FACs sorting. For each, 4 sorted populations of cells (p4, p5, p6, p7) were collected and these were subjected to microarray. A total of 12 samples: 3 replicates each of p4, p5, p6, and p7.

2010-04-08 | E-GEOD-13201 | biostudies-arrayexpress

Gene Expression Profiling of Cutaneous CD30+ Lymphoproliferative Disorders by RNA-seq

Project description:Cutaneous CD30+ lymphoproliferative disorder (CD30+LPDs), including lymphomatoid papulosis (LyP) and primary cutaneous anaplastic large-cell lymphoma (PCALCL), comprises the second most common group of cutaneous T cell lymphoma. Previously, we reported that special AT-rich sequence-binding protein1 (SATB1), a thymocyte specific chromatin organizer, was over-expressed and promoted malignant T-cell proliferation in a portion of CD30+LPDs, whereas other CD30+LPDs didn't express SATB1 at all. To elucidate the underlying molecular events in CD30+LPDs with differential SATB1 expression, we subjected 4 SATB1+ and 3 SATB1- CD30+LPDs skin biopsies to second-generation RNA-sequencing (RNA-seq). These data provide a significant resource for studies of CD30+LPDs.

2019-01-25 | GSE109620 | GEO

Membrane-polysome associated fractionation of HES2 embryonic stem cells

Project description:The membrane fraction and the cytosolic fraction of HES2 cells were collected and subjected to microarray. The experiment was performed in triplicate Keywords: membrane-polysome fractionation of RNA Three consecutive passages of HES2 cells were grown and subject to membrane-polysome fractionation. A total of 6 samples, (3 replicates of 2 fractions) were included in the experiment

2010-04-07 | E-GEOD-14037 | biostudies-arrayexpress

Characterization of human CD30+ B cells and their relationship to Hodgkin and Reed-Sternberg cells of Hodgkin lymphoma

Project description:Small subsets of B cells in the germinal center (GC) and in extrafollicular regions of lymph nodes express the activation marker CD30. Very little is known about the specific features of these cells and their relationship to the CD30-expressing Hodgkin and Reed/Sternberg (HRS) cells of Hodgkin lymphoma. Phenotypic and immunoglobulin V gene analyses revealed that CD30+ GC B lymphocytes represent typical GC B cells, and that CD30+ non-GC B cells are mostly post-GC B cells. However, despite these seemingly distinct identities, both CD30+ subsets share an unexpectedly large overlap in specific transcriptome patterns, and are strikingly different from bulk GC B cells and classical memory and plasma cells, respectively. A main common feature of these CD30+ B cells is a strong MYC signature. CD30+ GC B cells appear to represent the recently described MYC+ GC B cell subset of recirculating centrocytes at the stage of centroblast transition. CD30+ non-GC B cells rather represent highly activated and proliferating memory B cells, differentiating into plasma cells. Notably, CD30+ B cells were more similar in their transcriptome patterns to HRS cells than any other B cell subset investigated, suggesting that HRS cells may either derive from CD30+ B cells or acquired a similar activation signature. In comparison to CD30+ B cells and other lymphomas, HRS cells show a remarkable downregulation of genes regulating cell cycle, genomic stability and polyploidity, providing a potential explanation for the genomic instability and multinuclearity of HRS cells.

2018-06-13 | GSE83441 | GEO

Ascorbate effect on serum free cultured hESC

Project description:Ascorbate activates CD30 expression and causes widespread specific demethylation of the epigenome of serum free cultured hESC. The genetic and epigenetic integrity of human Embryonic Stem cells (hESCs) is critical to their future applications in research and medicine. hESC cultured in serum free media can accumulate point mutations, aneuploidy and progressive epigenetic changes over prolonged culture in vitro. We have identified ascorbate as the only molecule in the very widely used Knock-out serum replacement medium that is sufficient to induce expression of CD30, a biomarker for aneuploidy in hESCs. In fact, we show that hESC cultured in the presence of ascorbate for 20 passages not only display demethylation of the CD30 locus, but exhibit widespread and remarkably specific and consistent demethylation of 1,847 genes in both HES2 and HES3 cells. The specific ascorbate induced demethylation changes in the hESC epigenome, of which 86% are shared between the two lines, identify a subset of genes in hESC, including CD30, that are sensitive to serum free culture medium induced epigenetic changes. Experiment. HES2 cells were maintained on 20% Fetal calf serum on mouse embryonic fibroblast feeder layers with mechanical dissection. Using HES2 after 99 mechanical passages (P99), hESC were grown for 17-20 (+17 - +20) passages in the presence of 20% Knock-Out Serum Replacement (Invitrogen) either with (+ASC) or without (-ASC) Ascorbate with enzymatic culturing. RNA (FACs sorted for the presence of the pluripotent stem marker TG30) from three passages (17, 19 and 20) (replicates) for both +ASC and âASC samples were arrayed.

2010-06-23 | E-GEOD-16590 | biostudies-arrayexpress

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