Abstract:
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Communication n° 413 Many viral vectors have been designed for the purpose of delivering transgenes into a variety of cells and tissues from many species. The effective delivery of the therapeutic agent in human muscle cells, and itÕs sustained expression, are major components in the evaluation of the effectiveness of cell therapy. Our goal here, is to assess the potentiality, for every vector that we possess, to really translate these expectations. Human myoblasts, from primary cultures, were transducted by different serotypes of adenoassociated virus (rAAV2), by an adenovirus (rAd5), a retrovirus and a lentivirus. Both retrovirid×s were vsvg pseudotyped and rAAV were of serotypes 1,2 and 5. The transductions consisted of 8 hours incubations of with various viral preparations, ranging from 10E5 to 10E10 IU/ml or vgp/ml. Confluent cultures of myoblasts were set into differentiation for 4 days and transducted in the same conditions as the myoblasts. The expression of the reporter gene was observed 48 hours post-transduction. For the myoblasts, 4 x 10E9 vgp/ml adenofections and 3 x 10E6 IU/ml lentifections yield positive expressions of reporter gene for 95% of populations. Retroviral and rAAV2/2 transductions were positives for 52 and 45%, respectively. There was no reporter gene expression detectable in rAAV2 of serotypes 1 and 5. Adenofections on the differentiated cells shown identical results as for the myoblasts. All other vectors gave less than 5% or no positive expression at all. These results show that adenofection and lentifection are the only effective tools for the delivery of gene therapy, in human muscle cells dedicated to cell therapy.
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