The selected cells are muscle-derived “AC-133” precursors (Yvan Torrente, Milan). This material is difficult to isolate and to amplify ex vivo without losing its myogenic properties. The most significant advances are: - The preparation of AC-133 cells retaining their in vivo myo-regenerating capacity was reproduced for the first time in another lab, thru the utilization of another less expensive culture medium (EGM2 Lonza) (Jenny Morgan, London). - Quality control: the experimental laboratory processes of isolation and amplification of AC-133 cells derived from dog and human muscle have been optimized. A battery of tests have been standardized to control the quality of the cells at each step of the procedure (Genosafe, Evry). - Heterogeneity of AC-1133 cells: this important feature has been evidenced, even after starting from one single cell clone. It has an impact on cell viability, cell proliferation, and in vivo myoregenerating capacity. At the present time it is a major hurdle on the way to clinical trials (Yvan Torrente, Milan). - Therapeutic trials in GRMD dog: 6 animals received arterial infusion of autologous AC-133 cells harnessed with the su7opt exon skipping device vectorized in a lentivirus (108 cells via subclavian route + 108 cells via femoral route). Encouraging results were obtained both at biological and clinical levels. No side effects were observed. The degree of histological and 4 functional recovery varied significantly from one dog to the other, suggesting that there are “good” and “bad” responders. Unexpectedly there was no correlation with the degree of dystrophin rescue. Functional improvement was evaluated by the “6 min walk test”, ability to climb stairs, and attested by video recording. It is already possible to say that this strategy combining splicing and cell therapy is able to elicit a sustained muscle regeneration with progressive appearance of dystrophin (7% of dys positive fibers after 6 months). These animals are still alive, allowing one to pursue long term surveys. These results belong to the most important advances obtained in 2010
Autologous stem cell therapy combined with exon skipping / Y. Torrente. ((Intervento presentato al convegno Towards clinical trials for DMD using exon-skipping obtained by combined gene and cell therapy - ICE 2010 tenutosi a Principality of Monaco nel 2010.
Autologous stem cell therapy combined with exon skipping
Y. TorrentePrimo
2012
Abstract
The selected cells are muscle-derived “AC-133” precursors (Yvan Torrente, Milan). This material is difficult to isolate and to amplify ex vivo without losing its myogenic properties. The most significant advances are: - The preparation of AC-133 cells retaining their in vivo myo-regenerating capacity was reproduced for the first time in another lab, thru the utilization of another less expensive culture medium (EGM2 Lonza) (Jenny Morgan, London). - Quality control: the experimental laboratory processes of isolation and amplification of AC-133 cells derived from dog and human muscle have been optimized. A battery of tests have been standardized to control the quality of the cells at each step of the procedure (Genosafe, Evry). - Heterogeneity of AC-1133 cells: this important feature has been evidenced, even after starting from one single cell clone. It has an impact on cell viability, cell proliferation, and in vivo myoregenerating capacity. At the present time it is a major hurdle on the way to clinical trials (Yvan Torrente, Milan). - Therapeutic trials in GRMD dog: 6 animals received arterial infusion of autologous AC-133 cells harnessed with the su7opt exon skipping device vectorized in a lentivirus (108 cells via subclavian route + 108 cells via femoral route). Encouraging results were obtained both at biological and clinical levels. No side effects were observed. The degree of histological and 4 functional recovery varied significantly from one dog to the other, suggesting that there are “good” and “bad” responders. Unexpectedly there was no correlation with the degree of dystrophin rescue. Functional improvement was evaluated by the “6 min walk test”, ability to climb stairs, and attested by video recording. It is already possible to say that this strategy combining splicing and cell therapy is able to elicit a sustained muscle regeneration with progressive appearance of dystrophin (7% of dys positive fibers after 6 months). These animals are still alive, allowing one to pursue long term surveys. These results belong to the most important advances obtained in 2010
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