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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D-7037), 0.25 mg/ml E. coli tRNA (Sigma, cat no. R-1753), 0.125 mg/ml salmon
sperm DNA (Sigma, cat. no. D-7656), 0.25 mg/ml heparin (Sigma, cat, no. H-3393
) in DEPCtreated water). 4. For each coverslip, place 180 ul of hybridization ...
The nucleoside stock solution is made up by dissolving the following in distilled
water at 37°C; adenosine, 0.8 mg/ml, guanosine, 0.85 mg/ml, cytidine, 0.73 mg/
ml, uridine, 0.73 mg/ml; and thymidine, 0.24 mg/ml, stock is filter-sterilized.
Box 11.3 Additives for Serum-Free Medium Final Concentration Stock Solution
Concentration Transferrin, 100 ug/ml 10 mg/ml (in PBS) “holo” form Insulin, 5ug/
ml 2.5 mg/ml (in 5 mM HCl, keep at 4°C) Zn salt Putrescine, 16 ug/ml 1.6mg/ml (in
Recombinant NGF (available from a number of suppliers) is best stored at -80°C
in aliquots containing 1 mg/ml bovine serum albumin; consult the supplier's
specification sheet for further details. Should any residual nonneuronal cells
Dissociation Medium (10x DM stock solutions) Volume (ml) for Final
concentration (1x) Stock solutions 250 ml 10x DM 90 ... Ky/Mg Stock (10 mM
kynurenate and 100mMMg”) To 250-ml flask add 190 ml H20, 1 ml phenol red
stock, 1.75 ml 1 ...
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