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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A novel transfection method using a complex of plasmid DNA, transferrin,
polylysine, and inactivated adenovirus (Cotten et al., 1992; Wagner et al., 1992)
leads to very high efficiency transfection of cell lines, presumably by a
combination of ...
(A) The plasmid amplicon contains the HSV Origin of replication and packaging
site and the expression cassette, in this case lacz driven by the HSV E3 promoter
and followed by the SV40 polyadenylation signal. (B) The packaging method ...
One method developed in Frank Graham's laboratory (McGrory et al., 1988; Bett
et al., 1994; see also Becker et al., 1994) uses a shuttle plasmid together with
another large plasmid (originally p]M17) containing the entire adenovirus
The method of construction involves cotransfection of a plasmid containing cis-
acting replication sequences and the expression cassette with a nonpackaging
complementing AAV plasmid and then superinfection with a helper adenovirus.
The choice of plasmids and transfection procedures appropriate for PC12 cells
are discussed later. PC12 cells that take up the plasmid can express transiently
the gene product of interest. Over time, most such cells lose the plasmid but, ...
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