A novel way of removing quiescent astrocytes in a culture of subcortical neurons grown in a chemically defined medium.
A new method has been described for removing a very small number of contaminating astrocytes in neuronal cultures (derived from the septal-diagonal band region of 17-day-old embryonic rat brain) grown in a chemically defined medium. The proportion of these glial fibrillary acidic protein (GFAP)-positive cells was usually less than 1.5% up to 10 days, but thereafter their number increased rapidly reaching 10-15% by 22 days in vitro. A prolonged exposure to normally used concentration of cytosine arabinoside (Ara-C; 10 microM) was toxic to both astroglial and neuronal cells, while a brief treatment (48 h) with a low level (4 microM) of Ara-C failed to eliminate these astrocytes, as judged by glutamine synthetase activity and GFAP-positive cell count. However, these quiescent astroglial cells could be easily eliminated if they were induced to proliferate by epidermal growth factor before exposure to Ara-C. The combined treatment with these agents had no effect on the number of acetylcholinesterase-positive cells, and on the development of cholinergic and GABA-ergic neurons, as measured in terms of choline acetyltransferase and glutamate decarboxylase activity, respectively.
Patel
A J
AJ
MRC Developmental Neurobiology Unit, London, U.K.
Seaton
P
P
Hunt
A
A
D000818:Animals; D001253:Astrocytes / Q000166:cytology* / Q000187:drug effects; D001921:Brain / Q000166:cytology; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media; D003561:Cytarabine / Q000494:pharmacology; D004622:Embryo, Mammalian; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D005974:Glutamate-Ammonia Ligase / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D051381:Rats
D003470:Culture Media; D005904:Glial Fibrillary Acidic Protein; D003561:Cytarabine; D004815:Epidermal Growth Factor; D007770:L-Lactate Dehydrogenase; D005974:Glutamate-Ammonia Ligase