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 known to 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 cord, mesencephalon and cortex. Two methods of viral inoculation were compared: the first inoculation was performed on the cultured cell at 2, 3 or 6 days in vitro (DIV) whereas the second inoculation was performed on the cell suspensions before seeding. All the infected CNS cells has the same aspect as the control cultures. In the spinal cord and the mesencephalic cultures, the glial cells were preferentially infected, especially when the cells were inoculated at 6 DIV. In the cortical 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 cell suspension before seeding.
INSERM U. 336 Ecole Nationale Supéieure de Chimie, Montpellier, France.
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D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002452:Cell Count; D002478:Cells, Cultured; D002490:Central Nervous System / Q000166:cytology* / Q000196:embryology* / Q000201:enzymology / Q000821:virology; D002540:Cerebral Cortex / Q000166:cytology / Q000196:embryology / Q000201:enzymology / Q000821:virology; D005822:Genetic Vectors; D007150:Immunohistochemistry; D007763:Lac Operon; D008636:Mesencephalon / Q000166:cytology / Q000196:embryology / Q000201:enzymology / Q000821:virology; D009474:Neurons / Q000166:cytology* / Q000201:enzymology / Q000821:virology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000166:cytology / Q000196:embryology / Q000201:enzymology / Q000821:virology; D014162:Transfection; D001616:beta-Galactosidase / Q000378:metabolism
D001616:beta-Galactosidase