Post-mitotic, differentiated myotubes efficiently produce retroviral vector from hybrid adeno-retrovirus templates.
We have examined the ability of proliferating myoblasts and post-mitotic, differentiated myotubes to produce retroviral vector using hybrid adeno-retroviral vectors as templates. We show that production of retroviral vector from myoblasts peaks 48 h after adenoviral infection at 4.8 x 10(4) cfu/ml and is scarcely detectable by 96 h. Both fully and partially differentiated myotubes were able to generate a sustained increase in the levels of retroviral vector compared with myoblasts peaking 48 h at 1.4 x 10(5) cfu/ml and 1.8 x 10(5) cfu/ml, respectively. Addition of the cell cycle inhibitor aphidicolin (5 microg/ml) had no effect on the production of retroviral vector from fully differentiated myotubes, but resulted in an 80% increase in vector production from partially differentiated myotubes. Thus indicating that retroviral vector production is more efficient in post-mitotic myotubes and is independent of muscle cell cycle progression.
Roberts
M L
ML
Division of Biochemistry, School of Biological Sciences, Royal Holloway-University of London, Egham, Surrey, UK.
Athanasopoulos
T
T
Pohlschmidt
M
M
Duisit
G
G
Cosset
F L
FL
Dickson
G
G
D016475:3T3 Cells; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D016590:Aphidicolin / Q000494:pharmacology; D002454:Cell Differentiation; D002460:Cell Line; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics* / Q000494:pharmacology; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008856:Microscopy, Fluorescence; D018485:Muscle Fibers, Skeletal / Q000187:drug effects / Q000648:ultrastructure / Q000821:virology*; D020388:Muscular Dystrophy, Duchenne / Q000628:therapy*; D012190:Retroviridae / Q000235:genetics*
D000998:Antiviral Agents; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D016590:Aphidicolin