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| Status |
Public on Sep 13, 2019 |
| Title |
Gene expression profiling of CHIR99021-induced skeletal myogenesis in human pluripotent stem cells |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Human pluripotent stem cell- (hPSC)-derived skeletal muscle progenitors (SMP)—defined as PAX7-expressing cells with myogenic potential—can provide an abundant source of donor material for muscle stem cell therapy owing to the near-infinite replication potential of PSCs. As in vitro myogenesis is decoupled from in vivo timing and the 3D-embryo structure, it remains difficult to definitively characterize what stage or type of muscle is modeled in culture. Here, gene expression profiling is analyzed in hPSCs over a 50 day skeletal myogenesis protocol and compared to gene expression datasets of other hPSC-derived skeletal muscle and adult murine satellite cells. Furthermore, day 2 cultures differentiated with high or lower concentrations of CHIR99021 were contrasted.
Expression profiling of the 50 day time course identified successively expressed gene subsets involved in mesoderm/paraxial mesoderm induction, somitogenesis, and skeletal muscle commitment/formation which could be regulative by a putative cascade of transcription factors. Initiating differentiation with higher CHIR99021 concentrations resulted in significantly higher expression of MSGN1 and TGFB-superfamily genes, notably NODAL, resulting in enhanced paraxial mesoderm and reduced ectoderm/neuronal gene expression. Comparison to adult satellite cells revealed that genes expressed in day 50 cultures correlated better with those expressed by quiescent or early activated satellite cells, which have a greater therapeutic potential than late activated satellite cells. Day 50 cultures were similar to other hPSC-derived skeletal muscle and both expressed known and novel SMP surface proteins.
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| Overall design |
24 samples were analyzed, reflecting 8 experimental conditions with 3 biological replicates each. Day 0 samples were used as a reference in downstream analysis to establish altered levels of gene expression at the later stages of differentiation.
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| Contributor(s) |
Shelton M, Blais A |
| Citation(s) |
31560727 |
| |
| Submission date |
May 13, 2019 |
| Last update date |
May 10, 2023 |
| Contact name |
Alexandre Blais |
| E-mail(s) |
[email protected]
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| Organization name |
University of Ottawa
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| Street address |
451 Smyth Road
|
| City |
Ottawa |
| State/province |
ON |
| ZIP/Postal code |
K1H 8M5 |
| Country |
Canada |
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| Platforms (1) |
| GPL20844 |
Agilent-072363 SurePrint G3 Human GE v3 8x60K Microarray 039494 [Feature Number Version] |
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| Samples (24)
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| Relations |
| BioProject |
PRJNA542692 |
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