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Scalable Production of Mouse Erythroid Cells using an in vitro Embryoid Body differentiation System [ChIP-Seq]

ABSTRACT: Scalable Production of Mouse Erythroid Cells using an in vitro Embryoid Body differentiation System [ChIP-Seq]

PROVIDER: PRJNA764671 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15970918_1.fastq.gz	Fastqsanger.gz
	SRR15970918_2.fastq.gz	Fastqsanger.gz
	SRR15970919_1.fastq.gz	Fastqsanger.gz
	SRR15970919_2.fastq.gz	Fastqsanger.gz
	SRR15970920_1.fastq.gz	Fastqsanger.gz

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Scalable Production of Mouse Erythroid Cells using an in vitro Embryoid Body differentiation System

Project description:Scalable Production of Mouse Erythroid Cells using an in vitro Embryoid Body differentiation System

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Scalable Production of Mouse Erythroid Cells using an in vitro Embryoid Body differentiation System [ChIP-Seq]

Project description:Mouse embryonic stem (mES) cells can be manipulated ex-vivo to recapitulate the process of erythropoiesis, whereby multipotential haematopoietic stem cells undergo lineage specification, differentiation and maturation to produce functional erythroid cells. Although very useful for identifying specific progenitors and precursors, this cell system has not been fully exploited as a source of cells to analyse erythropoiesis, compared with primary erythroid cells or immortalised cell lines. Here we have established a protocol in which erythroid cells can be isolated in a scalable manner from differentiated embryoid bodies (EBs). Using transcriptional and epigenetic analysis, we show that all purified erythroid cells in this system are derived from the primitive haematopoietic lineage. Finally, we show that this system faithfully recapitulates normal erythropoiesis in the embryonic mouse and fully mimics the effects of natural and engineered mutations seen in full mouse models. Data deposited here comprises chromatin profiling of the EB derived CD71+ erythroid cells; chromatin accessibility (ATAC-Seq), characterisation of the open chromatin by associated histone modifications (ChIP-Seq), chromatin structure (CTCF ChIP-Seq and Capture C interaction data). WT CD71+ cells were compared to either mutant CD71+ cells or other erythroid tissues (mouse adult spleen, E13.5 fetal liver, early embryonic circulating blood) representing both definitve and primitive erythroid lineages.

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Scalable Production of Mouse Erythroid Cells using an in vitro Embryoid Body differentiation System [Capture-C]

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Scalable Production of Mouse Erythroid Cells using an in vitro Embryoid Body differentiation System [ATAC-Seq]

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Scalable Production of Mouse Erythroid Cells using an in vitro Embryoid Body differentiation System [ATAC-Seq]

Project description:Scalable Production of Mouse Erythroid Cells using an in vitro Embryoid Body differentiation System [ATAC-Seq]

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Scalable Production of Mouse Erythroid Cells using an in vitro Embryoid Body differentiation System [Capture-C]

Project description:Scalable Production of Mouse Erythroid Cells using an in vitro Embryoid Body differentiation System [Capture-C]

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Scalable Production of Mouse Erythroid Cells using an in vitro Embryoid Body differentiation System

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Early embryoid body differentiation

Project description:Embryonic stem (ES) cells, when grown in suspension culture without feeders, spontaneously form round structures known as embryoid bodies. Given the appropriate conditions, these cells can differentiate over time into precursors of all three germ layers. Embryoid bodies, in a disorganized way, mimic early embryonic development to a certain extent and can be used as a synchronously differentiating large scale source of tissue for the study of biological determinants of early differentiation. Embryoid bodies have been used as a source for most early protocols that derive specific differentiated cell types from undifferentiated ES cells, although some protocols, notably those that derive neurons from ES cells, have moved on from EBs as a result of varying replicability and yield. We have decided to look at the transcriptomic profiles of embryoid bodies during the initial stages of embryoid body formation and differentiation, in order to pinpoint novel determinants of key developmental stages. Keywords: time course

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Project description:Expression profiling of a 16 day mouse embryoid body (EB) differentiation time course.

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