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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Also, it is important to centrifuge antibody solutions to remove any precipitates
that may have formed during storage (we centrifuge at 10,000rpm for 10min in a
microfuge). Fine precipitates, either in the primary antibody solution or in labeled
15-ml clear conical centrifuge tubes and rack. 100-mm plastic Petri dishes. 10.
Coarse forceps. 11. Two pairs of watchmaker's forceps (Dumont no. 5). 12. One
small (4-inch) pointed dissecting scissors. 13. Medium-fine iridectomy scissors ...
DISSOCIATION AND PLATING When the dissection is completed transfer the
ganglia to a centrifuge tube containing 0.25% trypsin in HBSS with glucose.
Incubate the ganglia for 30min in a water bath at 37°C, then transfer them to a
Mince the tissue into small pieces with scissors, then transfer it to a 50-ml
centrifuge tube in a final volume of 12 ml of HBSS. Add 1.5 ml each of 2.5%
trypsin (Gibco BRL no. 25095-019) and 1% DNAse (Sigma no. DN-25) and
incubate at 37°C ...
Tap side of centrifuge tube to loosen cells and add 1 to 2 ml of the complement-
antibody mixture to cells. Resuspend cells in the complement-antibody mixture
and leave at 37°C for 30 min. To wash cells, top off tube with D-MEM-FCS and ...
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