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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Type I collagen was introduced as a substrate for neuronal cell culture by
Bornstein in 1958. As collagen is not found in brain, it was known at the time that
collagen was not a physiological substrate for CNS neurons, but it offered many ...
Box 6.1 Preparation and Application of Collagen A. Preparation of Rat Tail
Collagen Stock Collagen stock is prepared from adult rat tails by modifications of
the procedure of Bornstein (1958). The tails (freshly collected or stored until use
Neurons plated onto Matrigel or collagen sometimes migrate together to form
small clusters of cells; this rarely occurs on polyornithine or polylysine. According
to the manufacturer, Matrigel contains a number of different growth factors and, ...
Applying Substrate to Culture Dishes The core method that will be described is
spraying collagen onto the agarose to form tiny islands of adhesive substrate.
Spraying should be done in a sterile environment, such as a culturing hood.
Substratum Preparation COLLAGEN SUBSTRATUM The culture substratum for
long-term cultures must be durable, suitable for neuronal outgrowth, and
compatible with various types of nonneuronal cell attachment. When used by
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