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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The most commonly used viral vectors are derived from herpes simplex virus 1 (
HSV) or adenovirus, although adenoassociated virus, vaccinia virus, and semliki
forest virus (SFV) also Physical Methods have been adapted as neuron ...
An advantage of viral vectors is that they can be used to transfect neurons in vivo
with fairly high efficiency, and in fact most have been developed by labs
interested in potential applications in gene therapy. All these viral vectors can
Table 4.1 Wiral Wectors for Neuron Transfection HELPER MAXIMUM TITERS*
EXPRESSION CONSTRUCTION VIRAL VECTOR DEFECTIVE REQUIRED
GENOME INSERT (pfu/ml) TIME TIME R600mbinant Yes No 150 kb DNA >30 kb
Whereas HSV vectors yield high transfection efficiencies at a multiplicity of
infection (MOI) of 1, adenovirus vectors can require an MOI of 10 to 100 for
efficient transfection of neurons, perhaps because neurons are not a natural host
Figure 4.8 Considering all factors, defective adenovirus vectors still are one of the
most promising methods for neuron transfection. More recent improvements in
adenovirus vector construction include the generation of adenoviruses defective
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