Efficient and reproducible myogenic differentiation from human iPS cells: prospects for modeling Miyoshi Myopathy in vitro

PLoS One. 2013 Apr 23;8(4):e61540. doi: 10.1371/journal.pone.0061540. Print 2013.

Abstract

The establishment of human induced pluripotent stem cells (hiPSCs) has enabled the production of in vitro, patient-specific cell models of human disease. In vitro recreation of disease pathology from patient-derived hiPSCs depends on efficient differentiation protocols producing relevant adult cell types. However, myogenic differentiation of hiPSCs has faced obstacles, namely, low efficiency and/or poor reproducibility. Here, we report the rapid, efficient, and reproducible differentiation of hiPSCs into mature myocytes. We demonstrated that inducible expression of myogenic differentiation1 (MYOD1) in immature hiPSCs for at least 5 days drives cells along the myogenic lineage, with efficiencies reaching 70-90%. Myogenic differentiation driven by MYOD1 occurred even in immature, almost completely undifferentiated hiPSCs, without mesodermal transition. Myocytes induced in this manner reach maturity within 2 weeks of differentiation as assessed by marker gene expression and functional properties, including in vitro and in vivo cell fusion and twitching in response to electrical stimulation. Miyoshi Myopathy (MM) is a congenital distal myopathy caused by defective muscle membrane repair due to mutations in DYSFERLIN. Using our induced differentiation technique, we successfully recreated the pathological condition of MM in vitro, demonstrating defective membrane repair in hiPSC-derived myotubes from an MM patient and phenotypic rescue by expression of full-length DYSFERLIN (DYSF). These findings not only facilitate the pathological investigation of MM, but could potentially be applied in modeling of other human muscular diseases by using patient-derived hiPSCs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers / metabolism
  • Cell Differentiation / drug effects*
  • Cell Differentiation / genetics
  • Cells, Cultured
  • Distal Myopathies / genetics*
  • Distal Myopathies / metabolism
  • Distal Myopathies / pathology
  • Doxycycline / pharmacology
  • Dysferlin
  • Electric Stimulation
  • Gene Expression
  • Gene Expression Profiling
  • Genetic Vectors
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / drug effects
  • Induced Pluripotent Stem Cells / metabolism*
  • Membrane Proteins / genetics*
  • Membrane Proteins / metabolism
  • Mice
  • Mice, SCID
  • Models, Biological
  • Muscle Fibers, Skeletal / cytology
  • Muscle Fibers, Skeletal / drug effects
  • Muscle Fibers, Skeletal / metabolism*
  • Muscle Proteins / genetics*
  • Muscle Proteins / metabolism
  • Muscular Atrophy / genetics*
  • Muscular Atrophy / metabolism
  • Muscular Atrophy / pathology
  • MyoD Protein / genetics*
  • MyoD Protein / metabolism
  • Transfection

Substances

  • Biomarkers
  • DYSF protein, human
  • Dysferlin
  • Membrane Proteins
  • Muscle Proteins
  • MyoD Protein
  • MyoD1 myogenic differentiation protein
  • Doxycycline

Supplementary concepts

  • Miyoshi myopathy

Grants and funding

This research was supported in part by the FIRST Program, Scientific Research Grant No.22790284 (to HS) from the Japan Society for the Promotion of Science (JSPS), and a grant from the Leading Project of MEXT (to HS and AS). This research was also supported in part by grants from the Ministry of Health, Labor, and Welfare of Japan (to TE) and Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (to TE). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.