Culturing Nerve Cells
A do-it-yourself manual for culturing nerve cells, complete with recipes and protocols.
Because neurons and glia in culture are remarkably similar to those in situ, culture systems make it possible to identify significant cell interactions and to elucidate their mechanisms. This book is in many ways a do-it-yourself manual for culturing nerve cells, complete with recipes and protocols. But it also provides an understanding of the principles behind the protocols. In effect the contributors invite you into their labs and provide much of the information you would obtain from such a visit.The authors of the introductory chapters present the nuts-and-bolts principles of growing nerve cells. The authors of the following chapters discuss the culturing of specific cell types. They explain how their experimental goals have shaped their particular cell culture approach and the advantages and disadvantages of the cell culture systems they have developed. They provide detailed protocols and describe their cultures in practical terms, from when the cells are first plated through the various phases of their development.
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Basal Media Most media are based on balanced salt solutions to which are
added amino acids, vitamins, and other nutrients at concentrations roughly
similar to those found in serum. The composition of the media commonly used in
which contains a higher concentration of NaHCO3, was intended to be
equilibrated at 10% CO2, although lower levels sometimes are used. The
formulation of these basal media was based on studies of the growth of various
cell lines, but ...
at concentrations of 5 to 50 plM. In any use of antimitotics, the possibility of toxic
effects on neurons must be considered; use of the lowest possible effective
concentration is recommended. AraC, even at low concentrations, is cytotoxic to
There is no apparent toxicity of Lipofectin at the concentrations employed here.
Lipofectamine can be slightly toxic to PC12 cells and, accordingly, incubations
greater than 14 to 15 h are not recommended. A variety of additional Lipofection
The appropriate concentration of laminin can vary from lot to lot, but 20 pg/ml is a
good starting point. Our usual combination involves coating the culture surface
first with 500 ug/ ml PLYS as described, followed by 20 ug/ml laminin for 45min at
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