Development of gene expression signature for defining the cell potency of muscle derived stem cells (MDSC) from mice of diffferent genotypes
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE39765 | GEO | 2012/08/01
SECONDARY ACCESSION(S): PRJNA171616
REPOSITORIES: GEO
Similar Datasets
Project description:In order to determine the cell potency, by identification of genes responsible for pluri/multi potency, we performed a global gene expression profiling of MDSC isolated from five week old male wild type(WT), C57Bl6J and another hypertrophied musculature mouse genotype called myostatin null (Mstn-/-) mice using microarray analysis and compared this gene expression to that of a standard mouse ES cell line W4. Muscle derived stem cells (MDSC) were isolated from WT and Mstn null mice using an established preplate technique and compared with the gene expression signature of standard mouse ES cell line W4 Entire hindlimb muscles of five week old WT and Mstn null male mice were subjected to primary cultures using the well established preplate technique based on adhesion properties of cells for sequential isolation of preplate6(PP6) cells on day4. The PP6 cells are called so because they are slowest adhering and attach to the matrigel coated dishes on day4. PP6 have stem cell like qualities as established by previous workers and are hence also known as MDSC. We expanded the MDSCs from the our isolation sources mentioned above for a fortnight in media containing 10% fetal bovine serum, 10% horse serum,1% chcken embryo extract,1% Penstep and 0.1M-BM-5M M-NM-2mercaptoethanol.RNA was extracted by RNeasy minit kit from Qiagen as per manufacturer's instructions and subjected to RNA integreity check by RIN score followed by microarray analysis for global gene expression using Agilent platform. Also, in parallel we cultured a standard mouse ES cell line from the company Taconic named W4 on feeder free conditions, extraced RNA and performed microarray analysis.
2012-07-31 | E-GEOD-39765 | biostudies-arrayexpress
Project description:The gold standard for examining pluripotency of stem cells is to see whether cells can contribute to entire body of animals. Here we show that the increased frequency of Zscan4 activation in mouse ES cells not only enhances, but also maintains their developmental potency in long-term cell culture. As the potency increases, even a whole animal can be produced from a single ES cell injected into 4N blastocyst at unusually high success rate. Although Zscan4-activated cells express genes that are also expressed in 2-cell stage mouse embryos, transiently Zscan4-activated state of ES cells is not associated with the high potency of ES cells. It is thus concluded that ES cells acquire higher potency by going through transient Zscan4 activation state more frequently than the regular state. Taken together, our results indicate that frequent activation of Zscan4 can rejuvenate pluripotent stem cells.
2013-06-12 | GSE15604 | GEO
Project description:The gold standard for examining pluripotency of stem cells is to see whether cells can contribute to entire body of animals. Here we show that the increased frequency of Zscan4 activation in mouse ES cells not only enhances, but also maintains their developmental potency in long-term cell culture. As the potency increases, even a whole animal can be produced from a single ES cell injected into 4N blastocyst at unusually high success rate. Although Zscan4-activated cells express genes that are also expressed in 2-cell stage mouse embryos, transiently Zscan4-activated state of ES cells is not associated with the high potency of ES cells. It is thus concluded that ES cells acquire higher potency by going through transient Zscan4 activation state more frequently than the regular state. Taken together, our results indicate that frequent activation of Zscan4 can rejuvenate pluripotent stem cells.
2013-06-12 | GSE26162 | GEO
Project description:Myostatin (Mstn) knockout mice exhibit large increases in skeletal muscle mass. However, relatively few of the genes that mediate or modify MSTN effects are known. In this study, we performed co-expression network analysis using whole transcriptome microarray data from Mstn-null and wild-type mice to identify genes involved in important biological processes and pathways related to skeletal muscle and adipose development.Genes differentially expressed between wild-type and Mstn-null mice were identified at 13.5 d.p.c. and 35 days after birth (d35) and further analyzed for shared DNA motifs using DREME. Known and novel transcription factor binding sites involved in MSTN signal transduction were enriched in differentially expressed genes. The use of regulatory impact factor (RIF), phenotypic impact factor (PIF), and differential hubbing analyses with the partical correlation and information theory (PCIT) co-expression analysis methods identified both known and potentially novel regulators of muscle development. These results provide new details of how MSTN may mediate transcriptional regulation as well as insight into novel regulators of MSTN signal transduction that merit further study regarding their physiological roles in muscle and adipose development.
2014-11-11 | GSE63154 | GEO
Project description:The gold standard for examining pluripotency of stem cells is to see whether cells can contribute to entire body of animals. Here we show that the increased frequency of Zscan4 activation in mouse ES cells not only enhances, but also maintains their developmental potency in long-term cell culture. As the potency increases, even a whole animal can be produced from a single ES cell injected into 4N blastocyst at unusually high success rate. Although Zscan4-activated cells express genes that are also expressed in 2-cell stage mouse embryos, transiently Zscan4-activated state of ES cells is not associated with the high potency of ES cells. It is thus concluded that ES cells acquire higher potency by going through transient Zscan4 activation state more frequently than the regular state. Taken together, our results indicate that frequent activation of Zscan4 can rejuvenate pluripotent stem cells. MC1 ES cells (for pZscan4-Emerald transfection) and R26R6 ES cells (for pZscan4-Cre ERT2 transfection) were grown on gelatin in 6-well plate, 5x10^5cells in suspension were transfected with 1ug of linearized pZscan4-Emerald vector or pZscan4-Cre ERT2 vector using Effectene (QIAGEN) according manufacturer protocol, and plated in gelatin coated 100 mm dishes. Cells were selected with 5 ug/ml blasticidin, colonies were picked on the 8th day, expanded and frozen for further analysis. DNA microarray analysis of pZscan4-Emerald cells was carried out as described. In brief, universal Mouse Reference RNA (Stratagene) were labeled with Cy5-dye, mixed with Cy3-labeled samples, and used for hybridization on the NIA Mouse 44K Microarray v2.2 (manufactured by Agilent Technologies #014117). The intensity of each gene feature was extracted from scanned microarray images using Feature Extraction 9.5.1.1 software (Agilent Technologies). Microarray data analyses were carried out by using an application developed in-house to perform ANOVA and other analyses (NIA Array Analysis software; http://lgsun.grc.nia.nih.gov)
2013-06-12 | E-GEOD-15604 | biostudies-arrayexpress
Project description:The gold standard for examining pluripotency of stem cells is to see whether cells can contribute to entire body of animals. Here we show that the increased frequency of Zscan4 activation in mouse ES cells not only enhances, but also maintains their developmental potency in long-term cell culture. As the potency increases, even a whole animal can be produced from a single ES cell injected into 4N blastocyst at unusually high success rate. Although Zscan4-activated cells express genes that are also expressed in 2-cell stage mouse embryos, transiently Zscan4-activated state of ES cells is not associated with the high potency of ES cells. It is thus concluded that ES cells acquire higher potency by going through transient Zscan4 activation state more frequently than the regular state. Taken together, our results indicate that frequent activation of Zscan4 can rejuvenate pluripotent stem cells. Previously, we have shown that constitutive and strong expression of Zscan4 arrest the proliferation of ES cells (refs). We sought for the system to mimic the transient expression of Zscan4 and decided to use ERT2 - Tamoxifen-inducible system (refs). The system allows one to keep a transgene off in the absence of Tmx and turn on in the presence of Tmx at will (refs). We first made a plasmid construct pCAG-Zscan4-ERT2, in which ZSCAN4 ORF fused with ERT2 domain can be driven by a strong ubiquitous promoter CAG. To further investigate this, we transfected pCAG-Zscan4-ERT2 plasmid into an F1 hybrid (C57BL/6J vs. 129) ES cell line V6.5 and isolated multiple cell clones. Based on the qRT-PCR analysis of Zscan4 ORF, we found that cell clones showed a variety of Zscan4 expression levels. We selected the clone that showed the highest Zscan4 expression levels (clone #18) and a clone #2 with the background Zscan4 level.
2013-06-12 | E-GEOD-26162 | biostudies-arrayexpress
Project description:Ets2-null ES cells are defective in differentiating into cardiac myocytes, evidenced by the lack of spontaneous beating, cardiac transcription factors and structural proteins. With the Phalanx mouse Onearray v2, we compared the gene expression profiles of cells derived from Ets2-null and wildtype ES cells. Standard embryoid body culture protocol was used to induce differentiation of murine ES cells. On the 10th day, replicates of either Ets2-null or control wildtype EB culture were collected in Trizol for total RNA extraction. Microarray was preformed by Phalanx.
2012-06-30 | E-GEOD-29710 | biostudies-arrayexpress
Project description:Recent studies have reported the deleterious physiological and metabolic changes in Mstn-/- mice including impaired force generation and susceptibility to contraction-induced injury. Such observations have raised the concerns about the functional quality of the increased muscle resulting from therapeutic blockade of Mstn. Here we provide proteomic evidence to demonstrate that therapeutic Mstn inhibition has minimal effects on muscle proteome composition whereas the genetic ablation of Mstn induces marked changes. Furthermore, this study also represents the first proteomic analysis of the pharmacological blockage of the Mstn/activin receptor pathway being actively pursued as a potential therapy for multiple muscle wasting disorders.
2016-05-31 | PXD003747 | Pride
Project description:Chronic inflammation is known to associate with prostate cancer development, but how epithelium-associated cancer initiating events cross-talk to inflammatory cells during prostate cancer initiation and progression is largely unknown. Using the Pten null murine prostate cancer model, we show an expansion of Gr-1+CD11b+ myeloid-derived suppressor cells (MDSCs) occurring intra-prostatically immediately following epithelium-specific Pten deletion without expansion in hematopoietic tissues. This MDSC expansion is accompanied by sustained immune suppression. Prostatic Gr-1+CD11b+ cells, but not those isolated from the spleen of the same tumor bearing mice, suppress T cell proliferation and express high levels of Arginase 1 and iNOS. Mechanistically, loss of PTEN in the epithelium leads to a significant upregulation of genes within the inflammatory response and cytokine-cytokine receptor interaction pathways, including Csf-1 and IL-1β, two genes known to induce MDSC expansion and immunosuppressive activities. Treating Pten null mice with the selective CSF-1 receptor inhibitor GW2580 decreases MDSC infiltration and relieves the associated immunosuppressive phenotype. Our study indicates that epithelium-associated tumor initiating events trigger the secretion of inflammatory cytokines and promote localized MDSC expansion and immune suppression, thereby promoting tumor progression. Laser-capture microdissection was used to extract epithelial cells from murine benign and Pten null prostate tumors. Their transcript levels were measured by the custom Agilent 4x44K whole mouse genome expression oligonucleotide microarrays. Each cohort had three biological replicates.
2014-04-03 | E-GEOD-56469 | biostudies-arrayexpress
Project description:Chronic inflammation is known to associate with prostate cancer development, but how epithelium-associated cancer initiating events cross-talk to inflammatory cells during prostate cancer initiation and progression is largely unknown. Using the Pten null murine prostate cancer model, we show an expansion of Gr-1+CD11b+ myeloid-derived suppressor cells (MDSCs) occurring intra-prostatically immediately following epithelium-specific Pten deletion without expansion in hematopoietic tissues. This MDSC expansion is accompanied by sustained immune suppression. Prostatic Gr-1+CD11b+ cells, but not those isolated from the spleen of the same tumor bearing mice, suppress T cell proliferation and express high levels of Arginase 1 and iNOS. Mechanistically, loss of PTEN in the epithelium leads to a significant upregulation of genes within the inflammatory response and cytokine-cytokine receptor interaction pathways, including Csf-1 and IL-1β, two genes known to induce MDSC expansion and immunosuppressive activities. Treating Pten null mice with the selective CSF-1 receptor inhibitor GW2580 decreases MDSC infiltration and relieves the associated immunosuppressive phenotype. Our study indicates that epithelium-associated tumor initiating events trigger the secretion of inflammatory cytokines and promote localized MDSC expansion and immune suppression, thereby promoting tumor progression Custom Agilent 4x44K whole mouse genome expression oligonucleotide microarrays were used to measure transcript levels in murine Pten null prostate cancer cell lines. Benign mouse prostate epithelia as wild type control against cell lines. Each cohort had three biological replicates.
2014-04-03 | E-GEOD-56468 | biostudies-arrayexpress