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Series GSE100722 Query DataSets for GSE100722
Status Public on Apr 26, 2019
Title In vivo generation of post-infarct human cardiac muscle by laminin-promoted cardiovascular progenitors [HS1001 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. We show high reproducibility of the differentiation protocol using time-course bulk RNA sequencing developed from different hESC lines. 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 Human embryonic stem cells HS1001 were cultured in wells coated with LN-521 and LN-221 using RPMI1460 supplemented with B21 without insulin medium. Differentiation commenced when cells reached confluency. Small molecule inhibitor CHIR99021 (inhibitor of GSK3) was added at day 0 followed by IWP2 (inhibiot of Wnt production) at day 3. Cells continue to be cultured in RPMI1460 supplemented with B21 without insulin medium for 10 days with medium change every other day. After day 10, medium was changed to RPMI1460 supplemented with B21 CTS grade medium. RNA was extracted from triplicates wells at day 0, 1, 3, 11, 14, 20, 30 and 90 and from 5 wells at day 5, 7 and 9.
Contributor(s) Yap L, Moreno-Moral A, Petretto E, Tryggvason K
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Submission date Jul 03, 2017
Last update date May 15, 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 (34)
GSM2692027 1001_pluri_1
GSM2692028 1001_pluri_2
GSM2692029 1001_pluri_3
This SubSeries is part of SuperSeries:
GSE100725 In vivo generation of post-infarct human cardiac muscle by laminin-promoted cardiovascular progenitors
BioProject PRJNA392969
SRA SRP111058

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SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE100722_HS1001_timecourse_TPM.txt.gz 1.8 Mb (ftp)(http) TXT
GSE100722_HS1001_timecourse_gene_raw_counts.txt.gz 1.7 Mb (ftp)(http) TXT
GSE100722_HS1001_timecourse_samples_info.txt.gz 828 b (ftp)(http) TXT
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Raw data are available in SRA
Processed data are available on Series record

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