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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This solution then is used in place of the primary antibody. Finally, if specific cell
types in the culture lack the antigen, they can serve as internal controls for
nonspecific binding. To reduce nonspecific binding, antibody solutions always
Box 6.2 Assessment of Cell Number A. Procedure for Counting Nuclei To count
attached cells, remove medium and replace with working counting solution. The
amount of solution used will depend on cell number, Generally, it is useful to aim
Box 12.4 Dissection and Dissociation Solutions Dissection solution (100ml stock
): Component Volume (ml) Final Concentration (mM) Double-distilled water
91.175 1 MMgCl2 0.58 5.8 mM 1 M CaCl2 0.025 0.25 mM 0.5 M HEPES 0.32 1.6
Recipes Before starting the experiments, prepare the following solutions and
media. Filter all the solutions and media with 0.2-pum-pore-size filter units and
store them at 4°C unless otherwise specified. BME stock solution: Per 100ml:86
Dissolve factor in an appropriate volume of medium, normally at 10 ug/ml. Aliquot
the solution in small containers in volumes of 10, 20, 50, and 100 ul per container
, and so on. The volumes used should be chosen to yield enough growth factor ...
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