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Transcriptome expression pattern of E11.5 PGCs, P2 GSCs, and CD49f+GSCs.

ABSTRACT: The cell identities of CD49f+GSCs were further identified by comparing them with the E11.5 PGCs and P2 GSCs. The transcriptomic analysis revealed that the CD49f+GSCs had 1/3 similar genes profile to the E11.5 PGCs and P2 GSCs. Further gene ontology (GO) analysis demonstrated that the E11.5 PGCs, P2 GSCs, and CD49f+GSCs shared the partial similar gene expression profile of pluripotency regulation signaling pathway, PI3K-AKT signaling, chemokine signaling, and HIF-1 signaling.

ORGANISM(S): Mus musculus

PROVIDER: GSE106664 | GEO | 2017/11/09

SECONDARY ACCESSION(S): PRJNA417561

REPOSITORIES: GEO

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Project description:Central questions like cardiomyocyte subtype emergence during cardiogenesis or availability of cardiomyocyte subtypes for cell replacement therapy require selective identification and purification of atrial and ventricular cardiomyocytes. However, characterization and implementation of pure cardiomyocyte subtypes is still challenging due to technical limitations. Our aim was to identify surface markers enabling the selective detection and purification of atrial and ventricular cardiomyocytes from mouse hearts. In a surface marker screen we found differential expression of CD49f in atrial and ventricular embryonic cardiomyocytes (E13.5). By flow cytometry we could correlate a high CD49f expression with MLC-2a on the single cell level; a low CD49f expression corresponded to MLC-2v. Based on the persisting differential CD49f expression we developed purification protocols for cardiomyocytes subtypes from the developing mouse heart. Flow sorting of E15.5 hearts into ErbB-2+/CD49flow and ErbB-2+/CD49fhigh cells led to a selective depletion (CD49flow) or enrichment of MLC-2a+ cells (CD49fhigh). We found a corresponding CD49f-dependent distribution of MLC-2a when pre-enriched neonatal cardiomyocytes (P2) were flow-sorted into CD49flow and CD49fhigh. Atrial and ventricular identity was confirmed by expression profiling and patch clamp analysis of sorted embryonic hearts, which unequivocally demonstrated that the sorted cells were viable and functional. For the first time, we introduce a non-genetic, antibody-based approach to specifically isolate atrial and ventricular cardiomyocytes from mouse hearts of various developmental stages. This newly gained capability of obtaining highly pure, viable cells will facilitate in-depths characterization of the individual cellular subsets and will aid translational research and therapeutic applications. The dataset comprises four different cardiomyocytes subtypes from the developing mouse heart. Embryonic (E15.5) hearts were dissociated and flow-sorted into ErbB-2+/CD49flow and ErbB-2+/CD49fhigh cardiomyocytes. Neonatal (P2) hearts were dissociated, contaminating non-myocytes were removed by MACS depletion, and the purified cardiomyocytes were flow-sorted into CD49flow and CD49fhigh cardiomyocytes. Four biological replicates were available for each sample groups. Microarray analysis was conducted on the Agilent Whole Mouse Genome Oligo Microarray 8x60K platform.

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Transcriptome expression pattern of E11.5 PGCs, P2 GSCs, and CD49f+GSCs.

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