ADENOVIRAL-MEDIATED TRANSFECTION OF THE LAC-Z GENE INTO RAT DISSOCIATED EMBRYONIC CENTRAL-NERVOUS-SYSTEM CELLS BEFORE AND AFTER SEEDING

The adenovirus carrying a reporter gene-the Lac-Z gene-is well known t o infect central nervous system (CNS) cells in primary cell cultures. The percentage of infected neurons with respect to the total number of neurons was studied in primary dissociated cultures as a function of the day of inoculation and the age of three rat CNS cultures: spinal c ord, mesencephalon and cortex. Two methods of viral inoculation were c ompared: the first inoculation was performed on the cultured cells at 2, 3 or 6 days in vitro (DIV) whereas the second inoculation was perfo rmed on the cell suspensions before seeding. All the infected CNS cell s had the same aspect as the control cultures. In the spinal cord and the mesencephalic cultures, the glial cells were preferentially infect ed, especially when the cells were inoculated at 6 DIV. In the cortica l cultures, there were more infected neurons than infected glial cells . The number of CNS cells was lower when inoculation was performed at 6 DIV as compared with 3 DIV. Very few infected GABA cells were found in the cultures. A high percentage of infected neuronal cells relative to the total number of neuronal cells was found when infection of the three types of cultures was performed on the dissociated embryonic ce ll suspension before seeding. (C) 1998 ISDN. Published by Elsevier Sci ence Ltd.