Immortalizing central nervous system cells.
This unit presents methods for isolating clonal, neural-derived cell lines. One approach for isolating such neural cell lines involves a replication-deficient retrovirus encoding a specific oncogene and a selectable marker which are used to infect dissociated CNS cells dissected at a developmental stage at which the cell population of interest has not undergone its terminal mitotic division. Also presented is a method for cloning by limiting dilution, which may be necessary to obtain a pure population of cells. Following growth under appropriate selection conditions, clones are isolated and tested for their ability to differentiate with the desired phenotypic properties. A method is also provided for coating tissue culture dishes, which is necessary for successful culture of CNS neurons.
Whittemore
S R
SR
University of Miami School of Medicine, Miami, Florida, USA.
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002455:Cell Division / Q000235:genetics; D002461:Cell Line, Transformed / Q000166:cytology* / Q000502:physiology; D002469:Cell Separation; D002478:Cells, Cultured; D002490:Central Nervous System / Q000166:cytology* / Q000254:growth & development / Q000502:physiology*; D002999:Clone Cells; D005260:Female; D005822:Genetic Vectors; D051379:Mice; D009474:Neurons / Q000166:cytology* / Q000502:physiology*; D009857:Oncogenes / Q000502:physiology; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D051381:Rats