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Series GSE127955 Query DataSets for GSE127955
Status Public on Mar 20, 2019
Title Single-cell RNA-seq of human cardiovascular progenitor cells generated with a chemically defined, xeno-free laminin based differentiation protocol
Organism Homo sapiens
Experiment type Expression profiling by high throughput sequencing
Summary Regeneration of injured human heart muscle is limited and an unmet clinical need. There are no methods for the reproducible generation of clinical quality stem-cell-derived cardiovascular progenitors (CVPs). We identified laminin-221 (LN-221) as the most likely expressed cardiac laminin. We produced it as human recombinant protein, and showed that LN-221 promotes differentiation of pluripotent hESCs towards cardiomyocyte lineage and downregulates pluripotency and teratoma associated genes. We developed a chemically defined, xeno-free laminin-based differentiation protocol to generate CVPs. Single-cell RNA sequencing of CVPs derived from hESC lines supported reproducibility and identified three main progenitor subpopulations. These CVPs were transplanted into myocardial infarction mice, where heart function was measured by echocardiogram and human heart muscle bundle formation was identified histologically. This method may provide clinical quality cells for use in regenerative cardiology.
Overall design We generated four independent single-cell RNA-sequencing datasets with day 9 and 11 cardiovascular progenitor cells derived from HS1001 and H1 hES cells respectively. H1 and HS1001 cells were cultured for 9 and 11 days in wells coated with LN-521+221 from day 0. We used nutristem to maintain the cells. Differentiation commenced when cells reached confluency. Small molecule inhibitor CHIR99021 (inhibitor of GSK3) was added at day 4 followed by IWP2 (inhibitor of Wnt production) at day 7.
Contributor(s) Yap L, Moreno-Moral A, Petretto E, Tryggvason K
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Submission date Mar 06, 2019
Last update date Mar 21, 2019
Contact name Aida Moreno-Moral
Organization name Duke-NUS Medical School
Street address 8 College Road
City Singapore
ZIP/Postal code 169857
Country Singapore
Platforms (1)
GPL21290 Illumina HiSeq 3000 (Homo sapiens)
Samples (4)
GSM3657225 Day9_H1_1000_cells
GSM3657226 Day9_1001_1000_cells
GSM3657227 Day11_H1_1000_cells
This SubSeries is part of SuperSeries:
GSE100725 In vivo generation of post-infarct human cardiac muscle by laminin-promoted cardiovascular progenitors
BioProject PRJNA525863
SRA SRP187760

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Supplementary file Size Download File type/resource
GSE127955_CVPs_single_cell_sequencing_gene_raw_counts.txt.gz 9.9 Mb (ftp)(http) TXT
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Raw data are available in SRA
Processed data are available on Series record

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