Efficiency of a high-titer retroviral vector for gene transfer into skeletal myoblasts

The Journal of Thoracic and Cardiovascular Surgery
R M el OakleyM H Yacoub

Abstract

Genetic transformation of skeletal myoblasts for myocardial repair is dependent on an efficient gene transfer system that integrates the genes of interest into the genome of the target cell and its progeny. The aim of this investigation was to evaluate the use of a new retrovirally based gene transfer system for this purpose. MFGnlslacZ retroviral vector, packaged in high-titer, split-genome packaging cell line (FLYA4) was used to transduce the skeletal myoblast cell line L6. L6 cells, cultured in 10% fetal calf serum, were transduced with the MFGnlslacZ vector by means of filtered supernatant from FLYA4 cells. Transduced L6 cells were divided into four groups. Group I cells were fixed as myoblasts 3 days after transduction. Group II cells were allowed to differentiate into myotubes. Group III cells were split every 3 days for 4 months. Group IV cells were split as in group III but then allowed to differentiate into myotubes. All samples were fixed and stained for beta-galactosidase activity. The effects on gene transfer of transforming growth factor-beta, insulin-like growth factor-I, and platelet-derived growth factor were determined by spectrophotometric assay of beta-galactosidase activity in cells transduced in the presenc...Continue Reading

References

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Citations

Jun 1, 2001·The Annals of Thoracic Surgery·R M El OakleyM H Yacoub
Dec 17, 2004·Journal of Cellular and Molecular Medicine·A D BachR E Horch
Jan 5, 2011·Organogenesis·Carlo Alberto RossiPaolo De Coppi

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