Long-term histological follow-up of genetically modified myoblasts grafted into the brain

Although primary muscle cells have been used as intracerebral vehicles for transgene expression in the past, data concerning their long-term survival after grafting into the brain, and the reaction of the host tissue to their implantation are lacking. In order to study these aspects, we have implanted, into the brain, primary muscle cells infected ex vivo with recombinant retroviruses carrying the E. coli LacZ gene. The muscle cells were delivered stereotaxically into different areas of the brain of adult rats and the grafts were analyzed up to 105 days after implantation. Intraventricular implantations did not lead to surviving grafts. In contrast, myoblasts developed when they were grafted into gray or white matter regions. They appeared numerous during the first weeks, but decreased dramatically in number over time. Over months, the grafts appeared to fill up with collagen. Astrocytes elaborated a continuous glia limitans surrounding the implant. Blood vessels coming from the host tissue were found within the grafts. The blood-brain barrier was permanently disrupted within the transplants. beta-Galactosidase activity was abundant during the first weeks, but decreased to a very low level subsequently. This decrease paralleled that of the number of muscle cells. In conclusion, myoblasts transplanted into the adult brain survived only temporarily, which implies a transient transgene expression. In addition, before being eliminated, muscle cells were surrounded by a glia limitans, which may limit exchanges with the host tissue. Altogether, these results suggest that intracerebral transplantation of myoblasts may possibly provide a relevant vehicle only for short-term delivery of a gene product.