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Adenoviral‐mediated transfection of the lac‐z geneinto rat dissociated embryonic central nervous systemcells before and after seeding
Author(s) -
Cot Christine,
Privat Alain,
Levallois Christine
Publication year - 1998
Publication title -
international journal of developmental neuroscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.761
H-Index - 88
eISSN - 1873-474X
pISSN - 0736-5748
DOI - 10.1016/s0736-5748(97)00048-8
Subject(s) - biology , embryonic stem cell , central nervous system , spinal cord , cell culture , transfection , cell , in vitro , cell type , microbiology and biotechnology , virology , gene , neuroscience , genetics
Abstract The adenovirus carrying a reporter gene—the Lac‐Z gene—is well known toinfect central nervous system (CNS) cells in primary cell cultures. The percentage of infectedneurons with respect to the total number of neurons was studied in primary dissociated culturesas 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 firstinoculation was performed on the cultured cells at 2, 3 or 6 days in vitro (DIV) whereas the second inoculation was performed on the cell suspensions before seeding. All theinfected CNS cells had the same aspect as the control cultures. In the spinal cord and themesencephalic cultures, the glial cells were preferentially infected, especially when the cells wereinoculated at 6 DIV. In the cortical cultures, there were more infected neurons than infected glialcells. The number of CNS cells was lower when inoculation was performed at 6 DIV ascompared with 3 DIV. Very few infected GABA cells were found in the cultures. A highpercentage of infected neuronal cells relative to the total number of neuronal cells was foundwhen infection of the three types of cultures was performed on the dissociated embryonic cellsuspension before seeding.