Successive injections in mdx mice of myoblasts grown with bFGF

We studied the effects of single and repeated sets of injections in the same muscle of mdx mice of myoblasts grown with or without a high concentration 100 ng ml-1 of basic fibroblast growth factor (bFGF). The injected myoblasts were obtained from non-dystrophic transgenic mice expressing the beta-galactosidase gene under the control of a muscle-specific promoter. In these experiments, the host muscle was not irradiated to prevent muscle regeneration by host myoblasts. The host muscle was not damaged before myoblast transplantation with notexin, marcaïne or cold to trigger a regeneration-degeneration cycle. Without such pretreatments, the first set of injections of myoblasts grown without bFGF produced only 8% beta-galactosidase-positive and dystrophin-positive muscle fibers 1 month after transplantation. The percentage of muscle fibers containing the donor reporter gene increased, however, to 26% following a second set of injections in the same muscle. The percentage of muscle fibers expressing the donor reporter gene was significantly higher when the myoblasts were grown with a high dose of bFGF. Indeed the first set of injections produced 34% beta-gal-positive fibers while a second set of injections raised this percentage to 54%. In all cases, the percentage of dystrophin-positive fibers was similar to that of beta-gal-positive fibers. Therefore a high percentage of muscle fibers of donor origin can be obtained without preliminary damaging treatments of the mdx muscle when myoblasts grown with bFGF are injected several times. The effects of bFGF is not produced by increasing the percentage of myoblasts in a primary muscle culture since improvement of myoblast transplantation was obtained with a pure myoblast clone even with a lower concentration (10 ng ml-1) of bFGF.