Expansion for hematopoietic cells from umbilical cord blood is a strategy for use this cell source in clinic transplants, however, it is important to know about the genomic changes that can occur in expanded cells. In order to detect global expression profiles changes in hematopoietic stem and progenitors cells generated in vitro, we analyzed hematopoietics populations obtained by FACS in fresh from umbilical cord blood. HSC (fHSC) was defined as CD34+ CD38- CD71- CD45RA- Lin- and were cocultured with stromal cell line OP-9 plus FL, SCF, IL3, IL6, TPO, GMCSF and G-CSF by 7 days, after time we repurified HSC population by FACS using same immunophenotype (ivHSC). In other hand, fresh erythroid progenitors cells (fEPC) were identified as CD34+CD38+CD71+CD45RA- Lin- and fresh myeloid progenitors cells (fMPC) were identified as CD34+CD38+CD71-CD45RA+Lin-. In vitro progenitors cells (ivEPC and ivMPC) were obtained by culturing fHSC in Stemspan serum-free media plus SCF, TPO, IL6, FL and IL3 by 10 days, after time cells were repurified by FACS using same immunophenotype for fresh progenitors. In vitro generated cells were compared with their corresponding fresh population cells.
Functional Integrity and Gene Expression Profiles of Human Cord Blood-Derived Hematopoietic Stem and Progenitor Cells Generated In Vitro.
Specimen part
View SamplesWhile the hypothalamo-pituitary-adrenal axis (HPA) activates a general stress response by increasing glucocorticoid (Gc) synthesis, biological stress resulting from infections triggers the inflammatory response through production of cytokines. The pituitary gland integrates some of these signals by responding to the pro-inflammatory cytokines IL6 and LIF and to a negative Gc feedback loop. The present work used whole-genome approaches to define the LIF/STAT3 regulatory network and to delineate cross-talk between this pathway and Gc action. Genome-wide ChIP-chip identified 3 449 STAT3 binding sites, whereas 2 396 genes regulated by LIF and/or Gc were found by expression profiling. Surprisingly, LIF on its own changed expression of only 85 genes but the joint action of LIF and Gc potentiated the expression of more than a thousand genes. Accordingly, activation of both LIF and Gc pathways also potentiated STAT3 and GR recruitment to many STAT3 targets. Our analyses revealed an unexpected gene cluster that requires both stimuli for delayed activation: 83% of the genes in this cluster are involved in different cell defense mechanisms. Thus, stressors that trigger both general stress and inflammatory responses lead to activation of a stereotypic innate cellular defense response.
Regulatory network analyses reveal genome-wide potentiation of LIF signaling by glucocorticoids and define an innate cell defense response.
Specimen part, Time
View SamplesGut microbiota and the circadian clock are both key regulators of the metabolic processes. Although recent evidence points to the impact of the circadian clock on microbiota, gut microbiota effect on diurnal host gene expression remains elusive. A transcriptome analysis of germ-free mice reveals subtle changes in circadian clock gene expression. However, a lack of microbiome leads to liver feminization and alters the expression of male-specific genes involved in lipid metabolism and xenobiotic detoxification associated with sustained activation of the Growth Hormone pathway. These results emphasize the mutual interaction of gut microbiota and its host even on unexpected functions. Overall design: Total RNA-Seq of testis and ovaries of conventional raised (convR) and Germ-free (GF) female mice under ad libitum feeding regime.
The Mouse Microbiome Is Required for Sex-Specific Diurnal Rhythms of Gene Expression and Metabolism.
Sex, Specimen part, Cell line, Subject
View SamplesGut microbiota and the circadian clock are both key regulators of the metabolic processes. Although recent evidence points to the impact of the circadian clock on microbiota, gut microbiota effect on diurnal host gene expression remains elusive. A transcriptome analysis of germ-free mice reveals subtle changes in circadian clock gene expression. However, a lack of microbiome leads to liver feminization and alters the expression of male-specific genes involved in lipid metabolism and xenobiotic detoxification associated with sustained activation of the Growth Hormone pathway. These results emphasize the mutual interaction of gut microbiota and its host even on unexpected functions. Overall design: Total RNA-Seq of primary hepatocytes treated with serum of conventionally raised (convR) and Germ-free (GF) male and female mice.
The Mouse Microbiome Is Required for Sex-Specific Diurnal Rhythms of Gene Expression and Metabolism.
Specimen part, Subject
View SamplesGut microbiota and the circadian clock are both key regulators of the metabolic processes. Although recent evidence points to the impact of the circadian clock on microbiota, gut microbiota effect on diurnal host gene expression remains elusive. A transcriptome analysis of germ-free mice reveals subtle changes in circadian clock gene expression. However, a lack of microbiome leads to liver feminization and alters the expression of male-specific genes involved in lipid metabolism and xenobiotic detoxification associated with sustained activation of the Growth Hormone pathway. These results emphasize the mutual interaction of gut microbiota and its host even on unexpected functions. Overall design: Total RNA-Seq of Germ-free (GF) male mice liver injected with ghrelin.
The Mouse Microbiome Is Required for Sex-Specific Diurnal Rhythms of Gene Expression and Metabolism.
Sex, Specimen part, Cell line, Treatment, Subject
View SamplesGut microbiota and the circadian clock are both key regulators of the metabolic processes. Although recent evidence points to the impact of the circadian clock on microbiota, gut microbiota effect on diurnal host gene expression remains elusive. A transcriptome analysis of germ-free mice reveals subtle changes in circadian clock gene expression. However, a lack of microbiome leads to liver feminization and alters the expression of male-specific genes involved in lipid metabolism and xenobiotic detoxification associated with sustained activation of the Growth Hormone pathway. These results emphasize the mutual interaction of gut microbiota and its host even on unexpected functions. Overall design: Total RNA-Seq of Germ-free (GF) male mice liver injected with growth hormone.
The Mouse Microbiome Is Required for Sex-Specific Diurnal Rhythms of Gene Expression and Metabolism.
Sex, Specimen part, Cell line, Treatment, Subject
View SamplesPitx1, critical regulator of a limited hindlimb-specific gene network, targets the limb development program common to both fore- and hindlimbs in order to implement hindlimb-specific limb morphology. Overall design: The gene regulatory networks governing forelimb vs. hindlimb development in mouse were investigated using expressing profiling of morphologically stage-matched e10.5 forelimbs and e11.0 hindlimbs, ChIPseq of chromatin marks, and ChIPseq of limb-specific transcription factors Pitx1 and Tbx5. The makeup of the Pitx1-directed components of the hindlimb gene network were investigated using expression profiling of Pitx1 null hindlimbs at two stages (e11.0 and e11.5).
Regulatory integration of Hox factor activity with T-box factors in limb development.
Specimen part, Cell line, Subject
View SamplesDeployment of a cell-specifying enhancer repertoire by the pioneer factor Pax7 The establishment and maintenance of cell identity depends on implementation of stable cell-specific chromatin landscapes. Pioneer transcription factors establish new cell fate competences by triggering chromatin remodeling during development. Here, we used pituitary cell specification to define the salient features of pioneer action. Comparison of purified pituitary cells of different lineages showed that chromatin accessibility differs at enhancers rather than promoters. The pioneer factor Pax7 specifies one pituitary lineage identity by opening a specific repertoire of enhancers that are distinct from the myogenic targets of Pax7. Pax7 binds its pioneer targets rapidly and days before chromatin remodeling and gene activation. Finally, enhancers opened by Pax7-dependent chromatin remodeling exhibit loss of DNA methylation and they acquire long term epigenetic memory. The present work identifies enhancer pioneering as a critical feature for cell fate specification and maintenance. Overall design: RNA extraction followed by high throughput sequencing (RNA-seq)
Pioneer factor Pax7 deploys a stable enhancer repertoire for specification of cell fate.
Specimen part, Cell line, Treatment, Subject
View SamplesThe combinatorial expression of the Hox genes along the body axes, referred to as the HOX code, is a major determinant of cell fate and plays a prevailing role in generating the animal body plan. In developing limb buds, the paralogous group 13 genes of the HoxA and HoxD clusters are essential for patterning the distal-most limb structures, the digits. Inactivation of HOXA13 and HOXD13 transcription factors (HOX13) leads to complete digit agenesis in mice, but how HOX13 regulate transcriptional outcomes and confer identity to the distal-most limb cells has remained elusive. Here we performed genome-wide profiling of HOX13 by chromatin immunoprecipitation and analyzed the transcriptome and chromatin state of wild type early and late-distal limb buds, as well as Hoxa13-/-;Hoxd13-/- compound mutant limb buds. Our results show that inactivation of HOX13 impairs the activation and repression of putative cis-regulatory modules specific to the late-distal limb cells. Loss of HOX13 also disrupts the specific, spatial patterning of gene expression along the proximal-distal axis of the developing limb buds. These results show that proper termination of the early limb transcriptional program and activation of the late-distal limb program are coordinated by the dual action of HOX13 on cis-regulatory modules. Overall design: Totla mRNAs from dissected distal parts of e11.5 forelimb, of wild-type as well as Hoxa13-/-;Hoxd13-/- mice
Regulatory integration of Hox factor activity with T-box factors in limb development.
Specimen part, Cell line, Subject
View SamplesRenal failure is characterized by important biological changes resulting in profound pleomorphic physiological effects termed uremia, whose molecular causation is not well understood. The data was used to study gene expression changes in uremia using whole genome microarray analysis of peripheral blood from subjects with end-stage renal failure (n=63) and healthy controls (n=20) to obtain insight into the molecular and biological causation of this syndrome.
Alteration of human blood cell transcriptome in uremia.
Sex, Specimen part, Disease, Disease stage, Race
View Samples