Genomics

Dataset Information

41

Efficient Generation of Cardiac Purkinje Fiber-like Cells from ESCs by Activating cAMP Signaling


ABSTRACT: Analysis of gene expression of mESC-derived cardiac purkinje fiber-like cells at mRNA level. Dysfunction of the cardiac conduction system (CCS) significantly impacts pathogenesis of arrhythmia, a major cause of morbidity and mortality. Strategies to derive cardiac conduction cells including Purkinje fiber cells (PC) would facilitate models for mechanistic studies and drug discovery, and also provide new cellular materials for regenerative therapies. A high-throughput chemical screen using CCS:lacZ and Contactin2:eGFP (Cntn2:eGFP) reporter embryonic stem cell (ESC) lines was used to discover a small molecule, sodium nitroprusside (SN), that efficiently promotes the generation of cardiac cells that express gene profiles and generate action potentials of PC-like cells. Imaging and mechanistic studies suggest that SN promotes the generation of PC from cardiac progenitors initially expressing cardiac myosin heavy chain, and that it does so by activating cAMP signaling. These findings provide a novel strategy to derive scalable PC, along with insight into the ontogeny of CCS development. Overall design: Total RNA isolated from mESC-derived cardiac purkinje fiber-like cells. Mouse Contactin2:eGFP transduced with Lenti-viral aMHC: mCherry embryonic Stem Cells was used. Day 4 of differentiated cells were treated with Sodium Nitroprusside(SN). After 25 days differentiation, by using FACS, we could separate SN-induced cells into three populations: negative, αMhc:mCherry+ (MHC) and Cntn2:eGFP+/dim αMhc:mCherry+(GFP). We then sequenced mRNA from these three populations and found that Cntn2:eGFP+ cells express cardiac purkinje-fiber gene profiles.

INSTRUMENT(S): Illumina HiSeq 2500 (Mus musculus)

SUBMITTER: Todd Evans 

PROVIDER: GSE68025 | GEO | 2016-04-20

SECONDARY ACCESSION(S): PRJNA281593

REPOSITORIES: GEO

Dataset's files

Source:
Action DRS
GSE68025_All_gene_normalized.counts.txt.gz Txt
Items per page:
1 - 1 of 1
altmetric image

Publications

Efficient Generation of Cardiac Purkinje Cells from ESCs by Activating cAMP Signaling.

Tsai Su-Yi SY   Maass Karen K   Lu Jia J   Fishman Glenn I GI   Chen Shuibing S   Evans Todd T  

Stem cell reports 20150528 6


Dysfunction of the specialized cardiac conduction system (CCS) is associated with life-threatening arrhythmias. Strategies to derive CCS cells, including rare Purkinje cells (PCs), would facilitate models for mechanistic studies and drug discovery and also provide new cellular materials for regenerative therapies. A high-throughput chemical screen using CCS:lacz and Contactin2:egfp (Cntn2:egfp) reporter embryonic stem cell (ESC) lines was used to discover a small molecule, sodium nitroprusside (  ...[more]

Similar Datasets

1000-01-01 | S-EPMC4471825 | BioStudies
2010-01-01 | S-EPMC3068837 | BioStudies
1000-01-01 | S-EPMC4542521 | BioStudies
2011-01-01 | S-EPMC3247655 | BioStudies
2010-01-01 | S-EPMC2930621 | BioStudies
2019-01-01 | S-EPMC6675655 | BioStudies
2015-01-01 | S-EPMC4418548 | BioStudies
2019-08-01 | GSE132658 | GEO
2011-01-01 | S-EPMC3060803 | BioStudies
1000-01-01 | S-EPMC4726039 | BioStudies