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Comparison between mouse ES/iPS derived neurosphere and mouse primary culture of neurospheres obtained from fetal mouse ganglionic eminence

ABSTRACT: Recent reports have emphasized the pitfalls of iPSC technology including the potential for immunogenicity of transplanted cells. It is serious safety-related concern for iPSC-based cell therapy. However, preclinical data supporting the safety and efficacy of iPSCs are also accumulating. To address the concern of immunogenicity of ESCs/iPSCs or ESCs/iPSCs-derived neurospheres, global gene expression profiles were compared between undifferentiated mouse ESCs (EB3 line), mouse iPSCs (38C2 line), and ESC/iPSC-derived neurosphere and mouse primary culture of neurosphere obtained from fetal mouse ganglionic eminence. Mouse adult sklin fibroblast was used as a control. We used affymetrix microarrays to compare the global gene expression of neurospheres prepared several origins. Keywords: Expression profiling by array RNA extracted from neurospheres was hybridized to Affymetrix microarrays.

ORGANISM(S): Mus musculus

SUBMITTER: Kenji Yoshida

PROVIDER: E-GEOD-31725 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Comparison between mouse ES/iPS derived neurosphere and mouse primary culture of neurospheres obtained from fetal mouse ganglionic eminence

Project description:Recent reports have emphasized the pitfalls of iPSC technology including the potential for immunogenicity of transplanted cells. It is serious safety-related concern for iPSC-based cell therapy. However, preclinical data supporting the safety and efficacy of iPSCs are also accumulating. To address the concern of immunogenicity of ESCs/iPSCs or ESCs/iPSCs-derived neurospheres, global gene expression profiles were compared between undifferentiated mouse ESCs (EB3 line), mouse iPSCs (38C2 line), and ESC/iPSC-derived neurosphere and mouse primary culture of neurosphere obtained from fetal mouse ganglionic eminence. Mouse adult sklin fibroblast was used as a control. We used affymetrix microarrays to compare the global gene expression of neurospheres prepared several origins. Keywords: Expression profiling by array

2012-03-01 | GSE31725 | GEO

Quantitative profiling of axonal guidance proteins during the differentiation of human neurospheres

Project description:Axon guidance is required for the establishment of brain circuits. Whether much of the molecular basis of axon guidance is known from animal models, the molecular machinery coordinating axon growth and pathfinding in humans remains to be elucidated. The use of induced pluripotent stem cells (iPSC) from human donors has revolutionized in vitro studies of the human brain. iPSC can be differentiated into neuronal stem cells which can be used to generate neural tissue-like cultures, known as neurospheres, that reproduce, in many aspects, the cell types and molecules present in the brain. Here, we analyzed quantitative changes in the proteome of neurospheres during differentiation. Relative quantification was performed at early time points during differentiation using iTRAQ-based labeling and LC-MS/MS analysis. We identified 6438 proteins, from which 433 were downregulated and 479 were upregulated during differentiation. We show that human neurospheres have a molecular profile that correlates to the fetal brain. During differentiation, upregulated pathways are related to neuronal development and differentiation, cell adhesion and axonal guidance whereas cell proliferation pathways were downregulated. We developed a functional assay to check for neurite outgrowth in neurospheres and confirmed that neurite outgrowth potential is increased after 10 days of differentiation and is enhanced by increasing cyclic AMP levels. The proteins identified here represent a resource to monitor neurosphere differentiation and coupled to the neurite outgrowth assay can be used to functionally explore neurological disorders using human neurospheres as a model.

2021-02-25 | PXD024177 | Pride

Human induced pluripotent stem cells (iPSCs) derived from T-cell compared with that from adult dermal fibroblast (aHDF) [gene expression]

Project description:In this study we determine the transcriptional profile by microarray of iPSCs and iPSC-derived neurospheres generated from T-cells or aHDF by using direct neurosphere method. iPSCs and iPSC-derived tertiary neurospheres generated from T-cells or aHDF subjected to the gene expression microarray analysis.

2016-01-14 | E-GEOD-76830 | biostudies-arrayexpress

Human induced pluripotent stem cells (iPSCs) derived from T-cell compared with that from adult dermal fibroblast (aHDF) [gene expression]

Project description:In this study we determine the transcriptional profile by microarray of iPSCs and iPSC-derived neurospheres generated from T-cells or aHDF by using direct neurosphere method.

2016-01-14 | GSE76830 | GEO

Expression data from dexamethasone treated mouse embryonic neural progenitor/stem cells isolated from wild type C57Bl/6 or caveolin-1 knockout mice

Project description:Neurosphere cultures prepared from E14.5 mouse cerebral cortex at passage 3 were treated for 4 hours with 100 nM dexamethasone We used microarrays to detail the global program of dexamethasone regulated gene expression in embryonic neural progenitor/stem cells. Cerebral cortex was isolated from E14.5 mouse fetuses and cultured as neurospheres for 3 passages prior to treatment with 100 nM dexamethasone or ethanol vehicle for 4 hours.

2014-05-21 | E-GEOD-49804 | biostudies-arrayexpress

Expression data from directly induced neural stem cells

Project description:Induced pluripotent stem (iPS) cells give rise to neural stem cells, which are applicable for therapeutic transplantation in treatment of neural diseases. However, generation of neural stem cells from iPS cells requires a careful selection of safe iPS clones. We sought to determine whether direct induction of neural stem cells from partially reprogrammed somatic cells is able to generate safer cells rapidly. We have successfully established direct induction system from fibroblast to neural stem cells. To characterize these directly induced neural stem cells, Gene expression profiles were compared with iPS cell or ES cell-derived neurosphere. We used affymetrix microarrays to compare the global gene expression of neurospheres prepared several method. RNA extracted from neurospheres was hybridized to Affymetrix microarrays. The mouse strain used in this study except ES/iPS cells was C57BL/6.

2012-03-31 | E-GEOD-31598 | biostudies-arrayexpress

mRNA expression data from iPSCs, ntESCs and iPSC-nt-ESCs

2011-05-31 | GSE24705 | GEO

Expression analysis of neurospheres generated in vitro

Project description:Neurospheres generated in vitro were treated with non-epinephrine or potassium chloride. Gene expression analysis was then carried out to identify genes that are up or down regulated due to chemical treatement. The microarray data for the neurosphere samples was generated at Queensland Brain Institute. Briefly, RNA was isolated from neurospheres generated in presence of either N (norepinephrine) or K (potassium chloride)and converted to cDNA using Applause WT-Amp ST kit (NuGEN). Fragmentation and biotin labelling of the cDNA was carried out using Encore Biotin module (NuGEN) for Affymetrix GeneChip Mouse Gene 1.0 ST arrays. Labelled cDNA was hybridised to arrays in an oven at 45M-BM-0C. Hybridised arrays were scanned using the Affymetrix GeneChip Scanner and the scanned data (.CEL files) was transferred to the Partek Genomics Suite and data analysis was carried out as described in Narayanan RK, Mangelsdorf M, Panwar A, Butler TJ, Noakes PG, et al. (2013).

2014-06-20 | E-GEOD-57440 | biostudies-arrayexpress

Generation of CNS neural stem cells and PNS derivatives from neural crest derived peripheral stem cells [Dataset 1]

Project description:Neural crest-derived neural stem cells (NCSCs) from the embryonic PNS can be reprogrammed in neurosphere culture (NS) to rNCSCs that produce CNS progeny, including myelinating oligodendrocytes. Using global gene expression analysis we now demonstrate that rNCSCs completely lose their previous PNS characteristics and acquire the identity of neural stem cells derived from embryonic spinal cord (SCSCs). Reprogramming proceeds rapidly and results in a homogenous population of Olig2-, Sox3- and Lex-positive CNS stem cells. Low-level expression of pluripotency inducing genes Oct4, Nanog and Klf4 argues against a transient pluripotent state during reprogramming. The acquisition of CNS properties is prevented in the presence of BMP4 (BMP NCSCs) as shown by marker gene expression and the potential to produce PNS neurons and glia. In addition, genes characteristic for mesenchymal and perivascular progenitors are expressed, which suggests that BMP NCSCs are directed towards a pericyte progenitor/mesenchymal stem cell (MSC) fate. Adult NCSCs from mouse palate, an easily accessible source of adult NCSCs, display strikingly similar properties. They do not generate cells with CNS characteristics but lose the neural crest markers Sox10 and p75 and produce MSCs. These findings show that embryonic NCSCs acquire a full CNS identity in neurosphere culture. In contrast, MSCs are generated from adult pNCSCs and BMP NCSCs, which reveals that postmigratory NCSCs are a source for MSCs up to the adult stage. Affymetrix Mouse 430_2 arrays were used to compare the gene expression profiles of E12.5 mouse spinal cord-derived neurospheres (SCSCs) and E12.5 DRG-derived neurospheres, cultured in the absence (rNCSCs) or in the presence of BMP4 (BMP NCSCs).

2014-11-01 | E-GEOD-56999 | biostudies-arrayexpress

mRNA expression data from iPSCs, ntESCs and iPSC-nt-ESCs

Project description:We generated three kinds of genetically identical mouse reprogrammed cells: induced pluripotent stem cells (iPSCs), nuclear transfer embryonic stem cells (ntESCs) and iPSC-nt-ESCs that are established after successively reprogramming of iPSCs by nuclear transfer (NT). NtESCs show better developmental potential than iPSCs, whereas iPSC-nt-ESCs display worse developmental potential than iPSCs. We used microarrays to distinguish the gene expression differences among three pluriptoent stem cells and identified that imprinted genes had a similar expression pattern in iPSCs and iPSC-nt-ESCs. We sought to obtain genetic identical pluripotent stem cell lines in order to minimize genetic variations among different reprogrammed cells. To that end, we established a genetically homogenous secondary reprogramming system, in which mouse embryonic fibroblasts (MEFs) carrying doxycycline (dox)-inducible lentiviruses expressing Oct4, Sox2, Klf4 and c-Myc were isolated and used as donors for different reprogramming experiments. We generated iPSCs after plating MEFs in the presence of dox in ES culture conditions, and then derived ntESCs after transplantation of the nucleus from the same MEFs into enucleated oocyte. Furthermore, we successively reprogrammed iPSCs by means of NT and established a set of nt-iPSC lines. Pluripotent stem cells generated from different reprogramming strategies were for RNA extraction and hybridization on Affymetrix microarrays.

2011-05-31 | E-GEOD-24705 | biostudies-arrayexpress

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