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Gene transfer into canine myoblasts

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Abstract

We have developed and characterized cultures of healthy and dystrophic canine myoblasts for the evaluation of various gene transfer protocols. The number of desmin-positive myoblasts was elevated (>>80%) in cultures of myoblasts obtained from different muscle territories, the diaphragm muscle giving rise to the purest cultures. Myoblasts from dogs turned out to be a very convenient source of well transfectable and transducible cells. Transfection with plasmid DNA allowed efficient transgene expression (50% of β-galactosidase positive cells and about 375 ng luciferase/mg protein after transfection with a calcium phosphate-precipitated plasmid). Infection with high concentrations of adenoviral and retroviral vectors allowed transgene (β-galactosidase or mini-dystrophin) detection in about 75 to 90% of the canine cells. Therefore, primary dog myoblast cultures represent a useful in vitro model for viral and non-viral gene delivery, as well as for functional evaluation and cell grafting with applications in genetic diseases, vaccination or production of circulating therapeutic proteins.

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Authors and Affiliations

  1. TRANSGENE S.A., 11 rue de Molsheim, 67082, Strasbourg Cedex, France

    Serge Braun, Christine Thioudellet, Frédéric Perraud, Marie-Christine Claudepierre, Horst Homann, Monika Lusky, Majid Mehtali & Andrea Pavirani

  2. Ecole Nationale Vétérinaire d'Alfort, Unité Pédagogique et de Recherche de Médecine, 7 avenue du général de Gaulle, 94700, Maisons Alfort, France

    Catherine Escriou

  3. Dept. of Bioanal. Chemistry, Astra-Draco AB, PO Box 34, S-221 00, Lund, Sweden

    Reiner Bischoff

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  1. Serge Braun
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  2. Christine Thioudellet
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  3. Frédéric Perraud
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  4. Catherine Escriou
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  5. Marie-Christine Claudepierre
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  6. Horst Homann
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  7. Monika Lusky
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  8. Majid Mehtali
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  9. Reiner Bischoff
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  10. Andrea Pavirani
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Braun, S., Thioudellet, C., Perraud, F. et al. Gene transfer into canine myoblasts. Cytotechnology 30, 181–189 (1999). https://doi.org/10.1023/A:1008026913715

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  • DOI: https://doi.org/10.1023/A:1008026913715

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