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A Myogenic Double-Reporter Human Pluripotent Stem Cell Line Allows Prospective Isolation of Skeletal Muscle Progenitors.

Cell reports | 2018

Myogenic differentiation of human pluripotent stem cells (hPSCs) 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 double-reporter human embryonic stem cell (hESC) line was generated for PAX7/MYF5, allowing prospective readout. This strategy allowed pathway screen to define efficient myogenic induction in hPSCs. Next, surface marker screen allowed identification of CD10 and CD24 for purification of myogenic progenitors and exclusion of non-myogenic cells. CD10 expression was also identified on human satellite cells and skeletal muscle progenitors. In vitro and in vivo studies using transgene and/or reporter-free hPSCs further validated myogenic potential of the cells by formation of new fibers expressing human dystrophin as well as donor-derived satellite cells in NSG-mdx4Cv mice. This study provides biological insights for myogenic differentiation of hPSCs using a double-reporter cell resource and defines an improved myogenic differentiation and purification strategy.

Pubmed ID: 30428361 RIS Download

Research resources used in this publication

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Associated grants

  • Agency: NIAMS NIH HHS, United States
    Id: R01 AR068293
  • Agency: NIAMS NIH HHS, United States
    Id: R21 AR071583

Publication data is provided by the National Library of Medicine ® and PubMed ®. Data is retrieved from PubMed ® on a weekly schedule. For terms and conditions see the National Library of Medicine Terms and Conditions.

This is a list of tools and resources that we have found mentioned in this publication.


Purified anti-human CD10 (antibody)

RRID:AB_314913

This monoclonal targets CD10

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CD24 (Heat Stable Antigen) (antibody)

RRID:AB_2737796

This monoclonal targets CD24 (Heat Stable Antigen)

View all literature mentions

Myf-5 (C-20) (antibody)

RRID:AB_631994

This polyclonal targets Myf-5 (C-20)

View all literature mentions

Lamin A antibody [EPR4100] (antibody)

RRID:AB_10866185

This monoclonal targets Lamin A antibody [EPR4100]

View all literature mentions

CD271 (NGF Receptor) (antibody)

RRID:AB_2737657

This monoclonal targets CD271 (NGF Receptor)

View all literature mentions

APC anti-human erbB3/HER-3 (antibody)

RRID:AB_2099567

This monoclonal targets erbB3/HER-3

View all literature mentions

NOD.Cg-Prkdcscid Il2rg/SzJ (organism)

RRID:IMSR_JAX:005557

Mus musculus with name NOD.Cg-Prkdcscid Il2rg/SzJ from IMSR.

View all literature mentions

APC anti-human erbB3/HER-3 (antibody)

RRID:AB_2099567

This monoclonal targets erbB3/HER-3

View all literature mentions

NOD.Cg-Prkdcscid Il2rg/SzJ (organism)

RRID:IMSR_JAX:005557

Mus musculus with name NOD.Cg-Prkdcscid Il2rg/SzJ from IMSR.

View all literature mentions

APC anti-human erbB3/HER-3 (antibody)

RRID:AB_2099567

This monoclonal targets erbB3/HER-3

View all literature mentions

CD271 (NGF Receptor) (antibody)

RRID:AB_2737657

This monoclonal targets CD271 (NGF Receptor)

View all literature mentions

CD24 (Heat Stable Antigen) (antibody)

RRID:AB_2737796

This monoclonal targets CD24 (Heat Stable Antigen)

View all literature mentions

Purified anti-human CD10 (antibody)

RRID:AB_314913

This monoclonal targets CD10

View all literature mentions

Lamin A antibody [EPR4100] (antibody)

RRID:AB_10866185

This monoclonal targets Lamin A antibody [EPR4100]

View all literature mentions

Myf-5 (C-20) (antibody)

RRID:AB_631994

This polyclonal targets Myf-5 (C-20)

View all literature mentions

CD271 (NGF Receptor) (antibody)

RRID:AB_2737657

This monoclonal targets CD271 (NGF Receptor)

View all literature mentions

CD24 (Heat Stable Antigen) (antibody)

RRID:AB_2737796

This monoclonal targets CD24 (Heat Stable Antigen)

View all literature mentions

Myf-5 (C-20) (antibody)

RRID:AB_631994

This polyclonal targets Myf-5 (C-20)

View all literature mentions

Purified anti-human CD10 (antibody)

RRID:AB_314913

This monoclonal targets CD10

View all literature mentions

Lamin A antibody [EPR4100] (antibody)

RRID:AB_10866185

This monoclonal targets Lamin A antibody [EPR4100]

View all literature mentions