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Murine CD8+ T cells: in vitro-derived vs. thymocytes

ABSTRACT: Transcriptional profiling of mouse in-vitro derived CD8+ cells (obtained after culturing BM-HSCs isolated from B6 mice) with OP9-DL1 cells for 35 days were compared to CD8+ thymocytes isolated from 4-5 weeks old B6 mice. Our goal was to determine similarities and differences in gene expression profile between in vitro-derived CD8+ cells and CD8+ cells isolated from the thymus. The procedure of in vitro differentiation of HSC using OP9-DL1 cocultures was previously described by: Schmitt, T. M., and J. C. Zuniga-Pflucker. 2002. Induction of T cell development from hematopoietic progenitor cells by delta-like-1 in vitro. Immunity 17:749-756 and Holmes, R., and J. C. Zuniga-Pflucker. 2009. The OP9-DL1 system: generation of T-lymphocytes from embryonic or hematopoietic stem cells in vitro. Cold Spring Harb Protoc 2009:pdb prot5156.

ORGANISM(S): Mus musculus

PROVIDER: GSE37669 | GEO | 2012/09/05

SECONDARY ACCESSION(S): PRJNA162585

REPOSITORIES: GEO

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Murine CD8+ T cells: in vitro-derived vs. thymocytes

Project description:Transcriptional profiling of mouse in-vitro derived CD8+ cells (obtained after culturing BM-HSCs isolated from B6 mice) with OP9-DL1 cells for 35 days were compared to CD8+ thymocytes isolated from 4-5 weeks old B6 mice. Our goal was to determine similarities and differences in gene expression profile between in vitro-derived CD8+ cells and CD8+ cells isolated from the thymus. The procedure of in vitro differentiation of HSC using OP9-DL1 cocultures was previously described by: Schmitt, T. M., and J. C. Zuniga-Pflucker. 2002. Induction of T cell development from hematopoietic progenitor cells by delta-like-1 in vitro. Immunity 17:749-756 and Holmes, R., and J. C. Zuniga-Pflucker. 2009. The OP9-DL1 system: generation of T-lymphocytes from embryonic or hematopoietic stem cells in vitro. Cold Spring Harb Protoc 2009:pdb prot5156. Biological replicates: 2 different samples of in vitro-derived CD8+ T cells from day 35 were compared to 2 different samples of thymic CD8+ cells.

2012-09-05 | E-GEOD-37669 | biostudies-arrayexpress

A Notch ligand, Delta-like 1 functions as an adhesion molecule for mast cells

Project description:To assess the effect of over-expression of Delta-like 1 on OP9 cells on gene expression in terms of Notch signaling and adhesion molecules, gene expressions of OP9, OP9-GFP (carrying GFP gene) and OP9-DL1 (carrying GFP and DL1 gene) cells (Immunity 2002;17:749-756) were analyzed.

2010-10-01 | GSE22659 | GEO

A Notch ligand, Delta-like 1 functions as an adhesion molecule for mast cells

2010-10-01 | E-GEOD-22659 | biostudies-arrayexpress

Notch signaling facilitates in vitro generation of cross-presenting classical dendritic cells

Project description:The IRF8-dependent subset of classical dendritic cells (cDC), termed cDC1, is important for cross-priming cytotoxic T cell responses against pathogens and tumors. Culture of hematopoietic progenitors with DC growth factor Flt3 ligand (Flt3L) yields very few cDC1 (in humans) or only immature "cDC1-like" cells (in the mouse). We report that OP9 stromal cells expressing Notch ligand Delta-like 1 (OP9-DL1) optimize Flt3L-driven development of cDC1 from murine immortalized progenitors and primary bone marrow cells. Co-culture with OP9-DL1 induced IRF8-dependent cDC1 with the phenotype (CD103+ Dec205+ CD8α+) and expression profile resembling ex vivo cDC1. OP9-DL1-induced cDC1 showed preferential migration towards Ccr7 ligands in vitro and superior T cell cross-priming and antitumor vaccination in vivo. Co-culture with OP9-DL1 also greatly increased the yield of IRF8-dependent CD141+ cDC1 from human bone marrow progenitors cultured with Flt3L. Thus, Notch signaling optimizes cDC generation in vitro and yields authentic cDC1 for functional studies and therapeutic applications.

2018-06-05 | GSE110577 | GEO

Comparison of gene expression from thymocyte populations and equivalent OP9-DL1 cultured cells

2014-07-19 | GSE49039 | GEO

Comparison of gene expression from thymocyte populations and equivalent OP9-DL1 cultured cells

Project description:Comparison between ex vivo immature, mature and stimulated T cells and in vitro generated counterparts. The T cells generated in vitro were cultured on OP9-DL1 stroma supplied with growth factors. Three in vivo (Thymus DPCD3+, Thymus SP8, PBMC CD8 POST) and three in vitro (CCd14 DPCD3+, CCd20 SP8, CCd27 Post) samples were collected. T-cells were sorted to high purity, RNA was extracted and hybridized on Affymetrix microarrays. No technical replicates were used. The expression profiles of in vivo samples were then compared to the ones of in vitro samples.

2014-07-19 | E-GEOD-49039 | biostudies-arrayexpress

Vitamin C Promotes Positive Selection of Mouse T Cells

Project description:We discovered that vitamin C (ascorbic acid) is required for differentiation of mouse bone marrow-derived progenitor cells in T cell cultures based on OP9-DL1 stromal cells. This microarray dataset evaluates differential expression of genes in OP9-DL1 stromal cells as well as in lymphocytes developing in the cultures when a stabilized form of ascorbic acid (l-ascorbic acid 2-phosphate; pAsc) is added. Cultures were maintained in the absence of pAsc for 13 days, at which time 800 micromolar pAsc was added. Cells were harvested after 24 and 72 hours (24 hours only in the case of OP9-DL1 cells) and solubilized in Trizol.

2011-08-24 | GSE31596 | GEO

Time series micro RNA analysis of immature T-cells and reMAIT cells by microarrays

Project description:To explore the molecular mechanisms underlying selective differentiation of MAIT-derived iPSCs into reMAITs, we conducted micro RNAs profiling based on microarrays. We sampled reMAITs during the time course of differentiation from iPSCs as well as mature MAITs isolated from cord blood (CB MAITs). As control, we used immature T cells differentiated from hematopoietic stem cells (HSCs). For each of reMAITs and the immature T cells, we selected four time points: Start, Early, Middle, and Late. MAITs isolated from CB were reprogrammed to iPSCs (PMID:23523177). The MAIT-derived iPSCs were cultured on OP9, and CD34+ CD43+ cells were isolated. These CD34+ CD43+ precursor cells were furthered cultured on OP9/DL1 for re-differentiation into MAITs. Based on the observation of the reported surface antigen profiles (PMID:23523177), reMAITs were harvested at four different time points: day 0 (Start), day 4 (Early), day 7-10 (Middle), and after day 30 (Late). For the immature T cells, CD34+ cells were isolated from CB using CD34 MicroBead Kit (Miltenyi Biotech). These CD34+ HSCs were cultured on OP9/DL1 for differentiation into T cell lineage as previously described (PMID:15494433). Based on the surface antigen profiles, the immature T cells were harvested at four different time points: CD34+ cells at day 0 (Start), CD4- CD8- double negative cells at day 21 (Early) and day 40 (Midlle), and CD4+ CD8+ double positive cells after day 50 (Late). Total RNA was extracted from each sample using RNeasy Kit (Quiagen). For micro RNA analysis, RNA was labeled with Cy3-pCp by ligation, and subjected to analysis using Human miRNA Microarray Release 19.0 8x60K (Agilent).

2017-03-14 | GSE88936 | GEO

Vitamin C Promotes Positive Selection of Mouse T Cells

2011-08-23 | E-GEOD-31596 | biostudies-arrayexpress

RNA-seq of EBF1E271A-transduced progenitors

Project description:Gene expression analysis in Ebf1-/- lymphocyte progenitors complemented with EBF1wt or EBF1E271A and co-cultured with OP9-DL1 feeder cells.

2022-07-01 | GSE201528 | GEO

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