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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Three of the most prominent and important ECM constituents—laminin,
fibronectin, and collagen—are available commercially (Collaborative Research,
Indianapolis, IN; Boehringer Mannheim; Gibco/BRL, Gaithersburg, MD; and
and an increase in the total length of the processes extended. The effects of
laminin on neuritic growth appear to be specific for axons, whereas dendritic
elongation is not enhanced on this substrate (Lein and Higgins, 1989; Lein et al.,
Coating with Laminin Stock solutions of laminin (1 mg/ml, Gibco no. 23017-015)
are delivered in glass vials on dry ice. The solution should be thawed slowly at 4°
C because rapid thawing causes the laminin to precipitate. Thawed laminin can ...
Laminin (Collaborative Research, Bedford MA, no. 40232) comes as a frozen
solution containing 1 mg in variable amounts of 50mM TRIS-HCl pH 7.4, 150 mM
NaCl (always less than 1 ml). Bring the volume to 1 ml with ddH2O, to make a 1 ...
Cultures of pontine explants require coating glass surfaces with laminin and with
PLY.S. As for the PLYS, each lot of laminin must be tested with the particular cell
types or explants to be used in the actual experiment. The appropriate ...
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