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PAX7::GFP hESC-derived myogenic progenitor(MPC) cell gene expression profile change after primary transplantation into murine muscle

ABSTRACT: PAX7::GFP hESC-derived myogenic progenitor(MPC) cell gene expression profile change after primary transplantation into murine muscle

PROVIDER: PRJNA786491 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR17139493.fastq.gz	Fastqsanger.gz
	SRR17139494.fastq.gz	Fastqsanger.gz
	SRR17139495.fastq.gz	Fastqsanger.gz
	SRR17139496.fastq.gz	Fastqsanger.gz

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PAX7::GFP hESC-derived myogenic progenitor(MPC) cell gene expression profile change after primary transplantation into murine muscle

Project description:To understand the mechanism of why in vivo PAX7::GFP+ cells having better engraftment capability

2022-02-01 | GSE190233 | GEO

Bulk RNA-sequencing of unsorted and PAX7::GFP-sorted ESC-derived myogenic progenitors

Project description:Tissue engineering strategies that combine human pluripotent stem cell-derived myogenic progenitors (hPDMs) with advanced biomaterials provide promising tools for engineering 3D skeletal muscle grafts to model tissue development in vitro and promote muscle regeneration in vivo. We recently demonstrated (i) the potential for obtaining large numbers of hPDMs using a combination of two small molecules and (ii) the application of electrospun fibrin microfiber bundles for functional skeletal muscle restoration following volumetric muscle loss. In this study, we demonstrate that the biophysical cues provided by the microfiber bundles can induce unsorted hPDMs to form multinucleated myotubes that express desmin and myosin heavy chain. To reduce the batch-to-batch variability of these samples, we tested a genetic PAX7 reporter line (PAX7::GFP) to sort for more homogenous populations of hPDMs. RNA sequencing and gene set enrichment analyses confirmed that PAX7::GFP-sorted hPDMs exhibited higher expression of myogenic genes while unsorted hPDMs demonstrated increased expression of genes associated with embryonic, neural crest, and fibroadipogenic progenitor lineages. We tested engineered skeletal muscle grafts derived from either PAX7::GFP-sorted or unsorted hPDMs within in vivo skeletal muscle defects. The unsorted hPDM-derived grafts also exhibited greater fibrosis and more proinflammatory responses. In contrast, the PAX7::GFP-sorted groups had moderately high vascular infiltration, more implanted cell association with embryonic myosin heavy chain (eMHC) regions, and greater CD206:CD86 ratios compared to unsorted hPDMs and acellular fibers suggesting they induced more pro-regenerative microenvironments. These findings demonstrated some promise for the use of PAX7::GFP-sorted hPDMS on fibrin microfiber bundles and provided some insights for improving the cell-biomaterial system to stimulate more robust in vivo skeletal muscle regeneration.

2022-12-09 | GSE178784 | GEO

Identification of PAX7-induced transcriptional changes and PAX7 genomic binding during skeletal myogenic differentiation of H9 embryonic stem cells

Project description:Skeletal myogenic commitment of human pluripotent cells can be achieved by doxycycline-inducible expression of the transcription factor PAX7. To gain further insights on PAX7 function during this process, we performed a time course whole transcriptome analysis of differentiating H9 human embryonic stem cells from doxycycline-treated and untreated cultures. In addition, we identified the genomic binding of PAX7 in one of the selected time point (referred as PAX7+ proliferating myogenic progenitors).

2017-06-30 | GSE98976 | GEO

Differentiation of the human PAX7-positive myogenic precursors/satellite cell lineage in vitro

Project description:Satellite cells (SC) are muscle stem cells which can regenerate adult muscles upon injury. Most SC originate from PAX7+ myogenic precursors set aside during development. Although myogenesis has been studied in mouse and chicken embryos, little is known about human muscle development. Here, we report the generation of human induced pluripotent stem cell (iPSC) reporter lines in which fluorescent proteins have been introduced into the PAX7 and MYOG loci. We use single cell RNA sequencing to analyze the developmental trajectory of the iPSC-derived PAX7+ myogenic precursors. We show that the PAX7+ cells generated in culture can produce myofibers and self-renew in vitro and in vivo. Together, we demonstrate that cells exhibiting characteristics of human fetal satellite cells can be produced in vitro from iPSC, opening interesting avenues for muscular dystrophy cell therapy. This work provides significant insights into the development of the human myogenic lineage.

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Bulk RNA-sequencing of unsorted and PAX7::GFP-sorted ESC-derived myogenic progenitors

Project description:Bulk RNA-sequencing of unsorted and PAX7::GFP-sorted ESC-derived myogenic progenitors

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A myogenic double reporter human pluripotent stem cell line allows prospective isolation of skeletal muscle progenitors

Project description:Myogenic differentiation of iPSCs has been done by gene–overexpression or directed differentiation. However, viral integration, long-term culture and the presence of unwanted cells are the main obstacles. By using CRISPR/Cas9n, a novel double reporter hESC line was generated for PAX7/MYF5, allowing prospective readout. This strategy allowed pathway screen to define efficient myogenic induction in hESC/iPSCs. Next, surface marker screen allowed identification of CD10 and CD24 for purification of myogenic progenitors. CD10 expression was also confirmed on human satellite cells and muscle progenitors. In vivo studies using transgene/reporter-free hESC/iPSCs further validated myogenic potential of the cells by dystrophin restoration and satellite cell engraftment in NSG-mdx4cv mice. In addition, side-by-side in vitro and in vivo comparison proved superior specificity of CD10/CD24 compared to recently reported markers (ERBB3/NGFR). This study provides new biological insights for myogenic differentiation using a new cell resource and identifies CD10 as a potential myogenic marker.

2018-12-03 | GSE121154 | GEO

Expression data from differentiating mouse embryonic stem cells wild type and lacking functional Pax7 gene

Project description:Myogenic differentiation relies on Pax7 function. We used embryonic stem cells lacking functional Pax7 to follow its role in derivation of skeletal myoblasts. Microarray analysis allowed us to compare transcriptomes of undifferentiated and differentiating embryonic stem cells of two genotypes, i.e. Pax7+/+ and Pax7-/- at day 7 and 21 of culture. Undifferentiated mouse embryonic stem cells, embryiod bodies at day 7, and embryoid body outgrowths at day 21 of differentiation were collected for RNA extraction and hybridization on Affymetrix microarrays.

2016-02-26 | E-GEOD-66483 | biostudies-arrayexpress

Identification of Pax7 binding sites in mouse skeletal muscle progenitors

Project description:Using dox-inducible mouse ES cells, we generated myogenic progenitors expressing the transcription factor Pax7. These cells were used for determining the genomic occupancy of the transcription factor Pax7.

2020-10-31 | GSE147057 | GEO

Expression data from mouse embryonic fibroblasts wild type and lacking functional Pax7 gene

Project description:Myogenic differentiation relies on Pax7 function. We used mouse embryonic fibroblasts lacking functional Pax7 to follow its role in terminally differentiated cells. Microarray analysis allowed us to compare transcriptomes of mouse embryonic fibroblasts of two genotypes, i.e. Pax7+/+ and Pax7-/-.

2016-10-26 | GSE80658 | GEO

Myogenic Progenitor Cell Lineage Specification by CRISPR/Cas9-based Transcriptional Activators

Project description:Engineered CRISPR/Cas9-based transcriptional activators can potently and specifically activate endogenous fate-determining genes to direct differentiation of pluripotent stem cells. Here, we demonstrate that endogenous activation of the PAX7 transcription factor results in stable epigenetic remodeling and directly reprograms human pluripotent stem cells into skeletal myoblast progenitor cells. Compared to the exogenous overexpression of PAX7 cDNA, we find that endogenous activation results in the generation of more proliferative myogenic progenitors that can maintain PAX7 expression over multiple passages in serum-free conditions while preserving the capacity for terminal myogenic differentiation. Transplantation of human myogenic progenitors derived from endogenous activation of PAX7 into immunodeficient mice resulted in a greater number of human dystrophin+ myofibers compared to exogenous PAX7 overexpression. RNA-seq analysis also revealed transcriptome-wide differences between myogenic progenitors generated via CRISPR-based endogenous activation of PAX7 and exogenous PAX7 cDNA overexpression. These studies demonstrate the utility of CRISPR/Cas9-based transcriptional activators for progenitor cell specification and their potential for regenerative medicine.

2020-04-10 | GSE145575 | GEO

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