Retrovirus-mediated gene transfer into mouse cerebellar primary culture and its application to the neural transplantation

Abstract

To develop a procedure for the transplantation of genetically modified brain primary cells, we transplanted cultured mouse cerebellar cells infected with recombinant retroviruses into the cerebella of adult mice and examined the expression of introduced gene-products in the host cerebellum after transplantation. After infection of cultured cerebellar cells with recombinant retroviruses harboring chloramphenicol acetyltransferase (CAT) gene, we selected only virus-infected cells for transplantation by culturing the cells in medium containing G418 for 3 weeks. CAT was continuously expressed in the cultured cerebellar cells during the 3-week incubation, but by immunoblotting analysis with antiglial fibrillar acidic protein (GFAP) or antineurofilament protein (NFP) antiserum the population of cultured cerebellar cells was found to change during the incubation. Immunocytochemical analyses using anti-CAT antiserum demonstrated that the transplanted cell mass containing CAT-positive cells was detectable in the cerebellum up to 3 weeks, but not 3 months after the transplantation of G418-selected cells into the cerebella of 7-week-old mice.