Preparation and partial characterization of highly purified primary cultures of neurons and non-neuronal (glial) cells from embryonic chick cerebral hemispheres and several other regions of the nervous system.
Purified cultures of neurons and non-neuronal (glial) cells were prepared from the cerebral hemispheres of 10-day chick embryos by a method previously used for embryonic chick sympathetic ganglia 16. This technique separates these cell types on the basis of both: (1) differences in the adhesiveness of neurons and non-neuronal cells to a collagen substrate; and (2) the capacity of neurons to form homotypic aggregates. Purity of the cerebral non-neuronal cultures was determined to be greater than or equal to 99.5% by microscopic examination, while that of the cerebral neuronal cultures was only 92%. Modification of the technique by periodic redissociation of the neuronal aggregates during cell separation increased the purity of the neuronal cultures to greater than or equal to 97% as determined both by microscopic examination and by measurement of levels of butyrylcholinesterase, an enzyme present in the non-neuronal cells. Highly purified cultures of neurons were also prepared from the optic lobes of 10-day chick embryos (greater than or equal to 98%), but attempts to obtain non-neuronal cultures of reasonable density from this tissue were unsuccessful. In addition, highly purified non-neuronal cultures (greater than or equal to 99.5%) were prepared from the dorsal root ganglia of 12-day chick embryos, but cultures enriched with dorsal root neurons could only be partially purified (82%). Specific activity of butyrylcholinesterase in cerebral non-neuronal cells was found to vary inversely with the density of non-neuronal cells.
D000818:Animals; D001921:Brain / Q000166:cytology* / Q000201:enzymology; D002091:Butyrylcholinesterase / Q000378:metabolism; D002448:Cell Adhesion; D002449:Cell Aggregation; D002478:Cells, Cultured; D002642:Chick Embryo; D003470:Culture Media; D007700:Kinetics; D008855:Microscopy, Electron, Scanning; D009457:Neuroglia / Q000166:cytology* / Q000648:ultrastructure; D009474:Neurons / Q000166:cytology* / Q000648:ultrastructure
D003470:Culture Media; D002091:Butyrylcholinesterase