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Assessing and enhancing migration of human myogenic progenitors using directed iPS cell differentiation and advanced tissue modelling

ABSTRACT: Assessing and enhancing migration of human myogenic progenitors using directed iPS cell differentiation and advanced tissue modelling

PROVIDER: PRJEB43338 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR9769190_1.fastq.gz	Fastqsanger.gz
	ERR9769190_2.fastq.gz	Fastqsanger.gz
	ERR9769200_1.fastq.gz	Fastqsanger.gz
	ERR9769200_2.fastq.gz	Fastqsanger.gz
	ERR9769181_1.fastq.gz	Fastqsanger.gz

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Assessing and enhancing migration of human myogenic progenitors using directed iPS cell differentiation and advanced tissue modelling

Project description:Muscle satellite stem cells are responsible for skeletal muscle growth and regeneration. Despite their differentiation potential, human MuSCs have limited in vitro expansion and in vivo migration capacity, limiting their use in cell therapies for diseases affecting multiple skeletal muscles. Several protocols have been developed to derive MuSC-like progenitors from human induced pluripotent stem (iPS) cells (hiPSCs) to establish a source of myogenic cells with controllable proliferation and differentiation. However, current hiPSC myogenic derivatives also suffer from limitations of cell migration, ultimately delaying their clinical translation. Here we use a multi-disciplinary approach including bioinformatics and tissue engineering to show that DLL4 and PDGF-BB improve migration of hiPSC-derived myogenic progenitors. Transcriptomic analyses demonstrate that this property is conserved across species and multiple hiPSC lines, consistent with results from single cell motility profiling. Treated cells showed enhanced trans-endothelial migration in transwell assays. Finally, increased motility was detected in a novel humanised assay to study cell migration using 3D artificial muscles, harnessing advanced tissue modelling to move hiPSCs closer to future muscle gene and cell therapies.

| S-SCDT-EMM-2021-14526 | biostudies-other

Assessing and enhancing migration of human myogenic progenitors using directed iPS cell differentiation and advanced tissue modelling.

Project description:Muscle satellite stem cells (MuSCs) are responsible for skeletal muscle growth and regeneration. Despite their differentiation potential, human MuSCs have limited in vitro expansion and in vivo migration capacity, limiting their use in cell therapies for diseases affecting multiple skeletal muscles. Several protocols have been developed to derive MuSC-like progenitors from human induced pluripotent stem (iPS) cells (hiPSCs) to establish a source of myogenic cells with controllable proliferation and differentiation. However, current hiPSC myogenic derivatives also suffer from limitations of cell migration, ultimately delaying their clinical translation. Here we use a multi-disciplinary approach including bioinformatics and tissue engineering to show that DLL4 and PDGF-BB improve migration of hiPSC-derived myogenic progenitors. Transcriptomic analyses demonstrate that this property is conserved across species and multiple hiPSC lines, consistent with results from single cell motility profiling. Treated cells showed enhanced trans-endothelial migration in transwell assays. Finally, increased motility was detected in a novel humanised assay to study cell migration using 3D artificial muscles, harnessing advanced tissue modelling to move hiPSCs closer to future muscle gene and cell therapies.

| S-EPMC9549733 | biostudies-literature

Expression data from undifferentiated and iPS cells differentiated to a myogenic fate [hPAX7]

Project description:Here, we report the generation of human induced Pluripotent Stem (iPS) cell reporter line in which a venus fluorescent protein have been introduced into the PAX7 locus. We use microarrays to compare the transcriptome of PAX7-venus+ cells after 3 weeks of myogenic differentiation to that of undifferentiated iPS Satellite cells (SC) are muscle stem cells which can regenerate adult muscles upon injury. Most SC originate from PAX7-positive myogenic precursors set aside during development. While myogenesis has been studied in mouse and chicken embryos, little is known about human muscle development. Here, we use microarrays to compare the transcriptome of myogenic cells differentiated in vitro from human and mouse ES and iPS cells reporter cell lines. We generated fluorescent reporter lines in which a fluorescent protein was introduced into the loci coding for Pax7 (mouse ES and human iPS) or MYOG (human iPS). Mouse ES and human iPS cells were differentiated to the myogenic lineage in vitro for three weeks according to the protocols described in Chal et al, Nature Biotech 2015 and to Chal, Al Tanoury et al, Nature Protocols, 2016. Fluorescent Pax7 or MYOG-positive cells were FACS-sorted after 3-weeks of differentiation in vitro and we used Affymetrix microarrays to analyze the transcriptome of the purified mouse and human cell populations and compare it to the transcriptome of undifferentiated mouse ES or human iPS cells.

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Expression data from undifferentiated and iPS cells differentiated to a myogenic fate [MYOG]

Project description:Here, we report the generation of human induced Pluripotent Stem (iPS) cell reporter line in which a venus fluorescent protein have been introduced into the MYOGENIN (MYOG) locus. We use microarrays to compare the transcriptome of MYOG-venus+ cells after 3 weeks of myogenic differentiation to that of undifferentiated iPS Satellite cells (SC) are muscle stem cells which can regenerate adult muscles upon injury. Most SC originate from PAX7-positive myogenic precursors set aside during development. While myogenesis has been studied in mouse and chicken embryos, little is known about human muscle development. Here, we use microarrays to compare the transcriptome of myogenic cells differentiated in vitro from human and mouse ES and iPS cells reporter cell lines. We generated fluorescent reporter lines in which a fluorescent protein was introduced into the loci coding for Pax7 (mouse ES and human iPS) or MYOG (human iPS). Mouse ES and human iPS cells were differentiated to the myogenic lineage in vitro for three weeks according to the protocols described in Chal et al, Nature Biotech 2015 and to Chal, Al Tanoury et al, Nature Protocols, 2016. Fluorescent Pax7 or MYOG-positive cells were FACS-sorted after 3-weeks of differentiation in vitro and we used Affymetrix microarrays to analyze the transcriptome of the purified mouse and human cell populations and compare it to the transcriptome of undifferentiated mouse ES or human iPS cells.

2020-04-21 | GSE148994 | GEO

Expression data from undifferentiated and embryonic stem (ES) cells differentiated to a myogenic fate [mPAX7]

Project description:Here, we use microarrays to compare the transcriptome of mouse Pax7-GFP ES reporter cell line after 3 weeks of myogenic differentiation in vitro to that of undifferentiated ES Satellite cells (SC) are muscle stem cells which can regenerate adult muscles upon injury. Most SC originate from PAX7-positive myogenic precursors set aside during development. While myogenesis has been studied in mouse and chicken embryos, little is known about human muscle development. Here, we use microarrays to compare the transcriptome of myogenic cells differentiated in vitro from human and mouse ES and iPS cells reporter cell lines. We generated fluorescent reporter lines in which a fluorescent protein was introduced into the loci coding for Pax7 (mouse ES and human iPS) or MYOG (human iPS). Mouse ES and human iPS cells were differentiated to the myogenic lineage in vitro for three weeks according to the protocols described in Chal et al, Nature Biotech 2015 and to Chal, Al Tanoury et al, Nature Protocols, 2016. Fluorescent Pax7 or MYOG-positive cells were FACS-sorted after 3-weeks of differentiation in vitro and we used Affymetrix microarrays to analyze the transcriptome of the purified mouse and human cell populations and compare it to the transcriptome of undifferentiated mouse ES or human iPS cells.

2020-04-21 | GSE148993 | GEO

Global transcriptional characterization of differentiating non-human primate (cynomolgus) iPAX7 iPS cells and comparison with their human counterpart

Project description:Pluripotent stem (PS) cells enable the scalable production of tissue-specific derivatives with therapeutic potential for various clinical applications, including muscular dystrophies. Given the similarity to human counterparts, the non-human primate (NHP) is an ideal preclinical model to evaluate several questions, including delivery, biodistribution and immune response. While the generation of human induced PS (iPS) cell-derived myogenic progenitors is well established, there has been no data for NHP counterparts, probably due to the lack of an efficient system to differentiate NHP iPS cells towards the skeletal muscle lineage. Here we report the generation of three independent Macaca fascicularis iPS cell lines and their myogenic differentiation using PAX7 conditional expression. Whole transcriptome analysis confirmed the successful sequential induction of mesoderm, paraxial mesoderm, and myogenic lineages. NHP myogenic progenitors efficiently gave rise to myotubes under appropriate in vitro differentiation conditions and engrafted in vivo into TA muscles of NSG and FKRP-NSG mice. Lastly, we explored the pre-clinical potential of these NHP myogenic progenitors in a single wild-type NHP recipient, demonstrating engraftment and characterizing the interaction with the host immune response. These studies establish an NHP model system in which to study iPS cell-derived myogenic progenitors.

2023-05-03 | GSE228567 | GEO

Expression data from undifferentiated and iPS cells differentiated to a myogenic fate [MYOG]

Project description:Expression data from undifferentiated and iPS cells differentiated to a myogenic fate [MYOG]

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Expression data from undifferentiated and iPS cells differentiated to a myogenic fate [hPAX7]

Project description:Expression data from undifferentiated and iPS cells differentiated to a myogenic fate [hPAX7]

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Expression data from undifferentiated and iPS/ES cells differentiated to a myogenic fate

Project description:Expression data from undifferentiated and iPS/ES cells differentiated to a myogenic fate

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Transcriptome data for human ES, FORESKIN and FORESKIN-derived iPS (ES4SKIN) cells

Project description:Assessing relevant differences between human induced pluripotent stem (iPS) cells and human embryonic stem (ES) cells is important, given that such differences may impact their potential therapeutic use. We used microarray to profile the changes in global gene expression arising from the reprogramming of somatic cells to induced pluripotent stem cells.

2011-01-05 | GSE26451 | GEO

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