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A New Extensively Characterised Conditionally Immortal Muscle Cell-Line for Investigating Therapeutic Strategies in Muscular Dystrophies

Muses, S; Morgan, JE; Wells, DJ; (2011) A New Extensively Characterised Conditionally Immortal Muscle Cell-Line for Investigating Therapeutic Strategies in Muscular Dystrophies. PLOS ONE , 6 (9) , Article e24826. 10.1371/journal.pone.0024826. Green open access

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Abstract

A new conditionally immortal satellite cell-derived cell-line, H2K 2B4, was generated from the H2K(b)-tsA58 immortomouse. Under permissive conditions H2K 2B4 cells terminally differentiate in vitro to form uniform myotubes with a myogenic protein profile comparable with freshly isolated satellite cells. Following engraftment into immunodeficient dystrophin-deficient mice, H2K 2B4 cells regenerated host muscle with donor derived myofibres that persisted for at least 24 weeks, without forming tumours. These cells were readily transfectable using both retrovirus and the non-viral transfection methods and importantly upon transplantation, were able to reconstitute the satellite cell niche with functional donor derived satellite cells. Finally using the Class II DNA transposon, Sleeping Beauty, we successfully integrated a reporter plasmid into the genome of H2K 2B4 cells without hindering the myogenic differentiation. Overall, these data suggest that H2K 2B4 cells represent a readily transfectable stable cell-line in which to investigate future stem cell based therapies for muscle disease.

Type: Article
Title: A New Extensively Characterised Conditionally Immortal Muscle Cell-Line for Investigating Therapeutic Strategies in Muscular Dystrophies
Open access status: An open access version is available from UCL Discovery
DOI: 10.1371/journal.pone.0024826
Publisher version: http://dx.doi.org/10.1371/journal.pone.0024826
Language: English
Additional information: © 2011 Muses et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. This work was supported by a European Union grant “SyntheGeneDelivery” (FFP 6 STREP project number 018716). Jennifer E. Morgan is funded by a Wellcome Trust University Award. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Keywords: SLEEPING-BEAUTY-TRANSPOSON, SKELETAL-MUSCLE, SELF-RENEWAL, SATELLITE CELLS, GENE-TRANSFER, MOUSE MUSCLE, STEM-CELL, IN-VIVO, EXPRESSION, MYOBLASTS
UCL classification: UCL
UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health > Developmental Neurosciences Dept
URI: https://discovery.ucl.ac.uk/id/eprint/1325301
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