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Transcription profile of fetal human CD140a-defined OPC differentiation in vitro

ABSTRACT: Human fetal dissociates from 19-22 week gestational age were magnetically sorted for CD140a antigen. CD140a-defined OPCs were plated into serum free conditions and allowed to differentiate in the absence of growth factors or mitogens. RNA was extracted from cells immediately following isolation and every day for 4 days. To block differentiation, matched cells were cultured in the presence of PDGF-AA (10ng/ml). This treatment prevents the acquisition of O4-positive oligodendrocyte cell fate and delays MBP mRNA expression by human CD140a-sorted OPCs.

ORGANISM(S): Homo sapiens

PROVIDER: GSE36431 | GEO | 2016/07/01

SECONDARY ACCESSION(S): PRJNA153239

REPOSITORIES: GEO

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Transcription profile of fetal human CD140a-defined OPC differentiation in vitro

Project description:Human fetal dissociates from 19-22 week gestational age were magnetically sorted for CD140a antigen. CD140a-defined OPCs were plated into serum free conditions and allowed to differentiate in the absence of growth factors or mitogens. RNA was extracted from cells immediately following isolation and every day for 4 days. To block differentiation, matched cells were cultured in the presence of PDGF-AA (10ng/ml). This treatment prevents the acquisition of O4-positive oligodendrocyte cell fate and delays MBP mRNA expression by human CD140a-sorted OPCs. 29 samples, 4 time points, 2 media conditions, at least three biological replicates per time point and media condition

2016-07-01 | E-GEOD-36431 | biostudies-arrayexpress

Isolation of fetal human CD140a and O4-defined oligodendrocyte progenitor cells.

2015-03-04 | GSE32589 | GEO

Isolation of fetal human CD140a and O4-defined oligodendrocyte progenitor cells.

Project description:Human fetal tissue dissociates between 18-22wk gestational age were FACS sorted on the basis of CD140a (PDGFRA) and O4 antigens. Each population was profiled immediately following FACS. 20 samples, 4 groups (FACS-sorted cell populations), between 3 and 6 replicates in each group, each replicate is from a separate patient sample

2015-03-04 | E-GEOD-32589 | biostudies-arrayexpress

CD133/CD140a-Based Isolation of Distinct Human Multipotent Neural Progenitor Cells and Oligodendrocyte Progenitor Cells

Project description:The mechanisms underlying the specification of oligodendrocyte fate from multipotent neural progenitor cells (NPCs) in developing human brain are unknown. In this study, we sought to identify antigens sufficient to distinguish NPCs free from oligodendrocyte progenitor cells (OPCs). We investigated the potential overlap of NPC and OPC antigens using multicolor fluorescence-activated cell sorting (FACS) for CD133/PROM1, A2B5, and CD140a/PDGFaR antigens. Surprisingly, we found that CD133, but not A2B5, was capable of enriching for OLIG2 expression, Sox10 enhancer activity, and oligodendrocyte potential. As a subpopulation of CD133- positive cells expressed CD140a, we asked whether CD133 enriched bone fide NPCs regardless of CD140a expression. We found that CD133+CD140a- cells were highly enriched for neurosphere initiating cells and were multipotent. Importantly, when analyzed immediately following isolation, CD133+CD140a- NPCs lacked the capacity to generate oligodendrocytes. In contrast, CD133+CD140a+ cells were OLIG2-expressing OPCs capable of oligodendrocyte differentiation, but formed neurospheres with lower efficiency and were largely restricted to glial fate. Gene expression analysis further confirmed the stem cell nature of CD133+CD140a- cells. As human CD133+ cells comprised both NPCs and OPCs, CD133 expression alone cannot be considered a specific marker of the stem cell phenotype, but rather comprises a heterogeneous mix of glial restricted as well as multipotent neural precursors. In contrast, CD133/CD140a-based FACS permits the separation of defined progenitor populations and the study of neural stem and oligodendrocyte fate specification in the human brain. 12 samples, 4 groups (FACS-sorted cell populations),3 replicates in each group, each replicate is from a separate patient sample

2013-04-23 | E-GEOD-40676 | biostudies-arrayexpress

Transcription profiling of differentiation of Rattus norvegicus oligodendrocytes from their progenitor cells

Project description:Oligodendrocytes undergo extensive changes as they differentiate from progenitors into myelinating cells. To better understand the; molecular mechanisms underlying this transformation, we performed a comparative analysis using gene expression profiling of A2B5+; oligodendrocyte progenitors and O4+ oligodendrocytes. Cells were sort-purified ex vivo from postnatal rat brain using flow cytometry. Using Affymetrix microarrays, 1707 transcripts were identified with a more than twofold increase in expression inO4+oligodendrocytes. Many genes required for oligodendrocyte differentiation were upregulated in O4+ oligodendrocytes, including numerous genes encoding; myelin proteins. Transcriptional changes included genes required for cell adhesion, actin cytoskeleton regulation, and fatty acid and; cholesterol biosynthesis. At the O4+ stage, there was an increase in expression of a novel proline-rich transmembrane protein (Prmp). Localized to the plasma membrane, Prmp displays adhesive properties that may be important for linking the extracellular matrix to the; actin cytoskeleton. Together, our results highlight the usefulness of this discovery-driven experimental strategy to identify genes relevant; to oligodendrocyte differentiation and myelination. Experiment Overall Design: Whole brain dissociates were prepared from one litter of 10 male postnatal day 7 rat pups for each of the 5 A2B5 bioligcal replicates and the 4 O4+ bioligical replicates. Total RNA was extracted from single A2B5+ and single O4+ cells sorted directly from postnatal day7 rat whole brain dissociates using flow cytometry.

2007-11-29 | E-GEOD-5940 | biostudies-arrayexpress

CD133/CD140a-Based Isolation of Distinct Human Multipotent Neural Progenitor Cells and Oligodendrocyte Progenitor Cells

2013-04-23 | GSE40676 | GEO

Casaccia OPC differentiation in the presence of TSA

Project description:OPCs were differentiated for one day in the presence or absence of the HDAC inhibitor, Trichostatin A.

2011-06-30 | E-MEXP-3028 | biostudies-arrayexpress

Generation of oligodendroglial cells by direct lineage conversion

Project description:We report the generation of induced oligodendrocyte precursor cells (iOPCs) by direct lineage conversion. Forced expression of the three transcription factors Sox10, Olig2 and Zfp536 was sufficient to convert mouse and rat fibroblasts into iOPCs with morphologies and gene expression signatures that resemble OPCs. We compared the global gene expression pattern of iOPCs, fibroblasts, primary OPCs from the neonatal rat brain, and their differentiated progeny. We purified iOPCs by O4 immunopanning three weeks after infection and extracted total RNA. Acutely isolated rat cortical OPCs were either used directly for RNA extraction or expanded in mitogen-containing media for 24h before switching into differentiation medium, lacking PDGF/NT-3 and containing T3. Cells were harvested for microarray analysis 3 and 6 days after induction of differentiation.

2013-04-07 | E-GEOD-40421 | biostudies-arrayexpress

Homo sapiens

Project description:Isolation of fetal human CD140a and O4-defined oligodendrocyte progenitor cells.

| PRJNA147089 | ENA

OPC Cell Culture with LPC 18:1 or LPC 18:1 (both + C13-His)

Project description:Rat OPCs were incubated in the presence of LPC 18:1 or LPC 18:0 and C13-His. Cultures controls included no treatment and a positive control for the differentiation of OPCs. LPC 18:1 was administered at 10 uM while C13-His was administered at 10 uM. Samples were process for mass spectrometry to identify the incorporation of C13-His in newly synthesized proteins.

2022-02-17 | PXD025177 | Pride

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