en
Doctoral thesis
Open access
English

Lentivector-mediated transfer of Bmi-1 and telomerase in muscle satellite cells yields a Duchenne myoblast cell line with long-term genotypic and phenotypic stability

Defense date2006-03-10
Abstract

Conditionally immortalized human cells are valuable substrates for basic bilogic studies, as well as for the production of specific proteins and for the creation of bioartificial organs. We previously demonstrated that the lentivector-mediated transduction of immortalizing genes into human primary cells is an efficient method for obtaining such cell lines. Here, we used human muscle satellite cells as model targets to examine the impact of the transduced genes on the genotypic and phenotypic characteristics of the immortalized cells. The most commonly used immortalizing gene, the SV40 large T antigen (T-Ag), was extremely efficient at inducing the continuous growth of primary myoblasts, but the resulting cells rapidly accumulated major chromosomal aberrations and exhibited profound phenotypic changes. In contrast, the constitutive expression of telomerase and Bmi-1 in satellite cells from a control individual and from a patient suffering from Duchenne's muscular dystrophy yielded cell lines that remained diploid and conserved their growth factor dependence for proliferation. However, despite the absence of detectable cytogenetic abnormalities, clones derived from satellite cells of a control individual exhibited a differentiation block "in vitro". In contrast, a Duchenne-derived cell line exhibited all the phenotypic characteristics of its primary parent, including an ability to differentiate fully into myotubes when placed in proper culture conditions. This cell line should constitute a useful reagent for a wide range of studies aimed at this disease.

eng
Keywords
  • Immortalization of Duchenne
  • Muscular dystrophy
  • DMD
  • Human moyblasts
Citation (ISO format)
CUDRÉ-MAUROUX, Christophe. Lentivector-mediated transfer of Bmi-1 and telomerase in muscle satellite cells yields a Duchenne myoblast cell line with long-term genotypic and phenotypic stability. 2006. doi: 10.13097/archive-ouverte/unige:402
Main files (1)
Thesis
accessLevelPublic
Identifiers
1152views
1263downloads

Technical informations

Creation10/29/2008 11:47:12 AM
First validation10/29/2008 11:47:12 AM
Update time03/14/2023 2:58:28 PM
Status update03/14/2023 2:58:28 PM
Last indexation01/29/2024 6:35:16 PM
All rights reserved by Archive ouverte UNIGE and the University of GenevaunigeBlack