Adenovirus mediated gene transfer in organotypic brain slices

Abstract

A replication-defective adenovirus vector carrying the reporter gene encoding β-galactosidase was used to transfect organotypic slices maintained in culture for up to 1 month. Three different delivery systems were used to inoculate the viral solution, either into the culture medium, or directly onto the surface of the slices or by microinjection into the tissue. Using the two first paradigms β-galactosidase expressing cells were mostly of glial phenotype and distributed throughout the slices without any specific regional pattern. In contrast, microinjection of the adenovirus resulted in a large number of both infected neurones and glia, concentrated at the site of injection. This method thus appears to be able to circumvent some of the constraints and limitations associated within vivogene transfer.

Keywords

• adenovirus
• gene transfer
• β-galactosidase
• organotypic slice culture
• hippocampus
• substantia nigra