Reliable and versatile immortal muscle cell models from healthy and myotonic dystrophy type 1 primary human myoblasts

Boris Pantic, Doriana Borgia, Silvia Giunco, Adriana Malena, Tohru Kiyono, Sergio Salvatori, Anita De Rossi, Emiliano Giardina, Federica Sangiuolo, Elena Pegoraro, Lodovica Vergani, Annalisa Botta

Research output: Contribution to journalArticle

Abstract

Primary human skeletal muscle cells (hSkMCs) are invaluable tools for deciphering the basic molecular mechanisms of muscle-related biological processes and pathological alterations. Nevertheless, their use is quite restricted due to poor availability, short life span and variable purity of the cells during in vitro culture. Here, we evaluate a recently published method of hSkMCs immortalization, relying on ectopic expression of cyclin D1 (CCND1), cyclin-dependent kinase 4 (CDK4) and telomerase (TERT) in myoblasts from healthy donors (n=3) and myotonic dystrophy type 1 (DM1) patients (n=2). The efficacy to maintain the myogenic and non-transformed phenotype, as well as the main pathogenetic hallmarks of DM1, has been assessed. Combined expression of the three genes i) maintained the CD56(NCAM)-positive myoblast population and differentiation potential; ii) preserved the non-transformed phenotype and iii) maintained the CTG repeat length, amount of nuclear foci and aberrant alternative splicing in immortal muscle cells. Moreover, immortal hSkMCs displayed attractive additional features such as structural maturation of sarcomeres, persistence of Pax7-positive cells during differentiation and complete disappearance of nuclear foci following (CAG)7 antisense oligonucleotide (ASO) treatment. Overall, the CCND1, CDK4 and TERT immortalization yields versatile, reliable and extremely useful human muscle cell models to investigate the basic molecular features of human muscle cell biology, to elucidate the molecular pathogenetic mechanisms and to test new therapeutic approaches for DM1 in vitro.

Original languageEnglish
Pages (from-to)39-51
Number of pages13
JournalExperimental Cell Research
Volume342
Issue number1
DOIs
Publication statusPublished - Mar 1 2016

Keywords

  • ASOs
  • CDK4
  • Cyclin D1
  • Human muscle cell models
  • Immortalization
  • Myotonic dystrophy type 1
  • TERT

ASJC Scopus subject areas

  • Cell Biology

Fingerprint Dive into the research topics of 'Reliable and versatile immortal muscle cell models from healthy and myotonic dystrophy type 1 primary human myoblasts'. Together they form a unique fingerprint.

  • Cite this

    Pantic, B., Borgia, D., Giunco, S., Malena, A., Kiyono, T., Salvatori, S., De Rossi, A., Giardina, E., Sangiuolo, F., Pegoraro, E., Vergani, L., & Botta, A. (2016). Reliable and versatile immortal muscle cell models from healthy and myotonic dystrophy type 1 primary human myoblasts. Experimental Cell Research, 342(1), 39-51. https://doi.org/10.1016/j.yexcr.2016.02.013