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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These technical problems were overcome by using agarose as a substrate.
Agarose is virtually nonadhesive for glial cells (Costăchel et al., 1969; Grinnell et
al., 1972; Westermark et al., 1978; Pontén and Stolt, 1980). Why cells avoid
Preparing Dishes The goal in preparing dishes is to have a surface with islands
of substrate on a background of agarose. In the method described here, glass
coverslips are attached to plastic dishes that have a central well cut out of the
It is important to avoid plastic fragments in the dishes, because the fragments can
damage the agarose surface. Attaching Coverslips with Sylgard Coverslips with
the agarose coating are attached to the plastic Petri dishes by Sylgard, ...
In addition to the preceding general comments, several problems are specific to
the microculture technique. Agarose detaching: Try being more careful to avoid
dust and gelled agarose in placing the agarose layer on coverslips. Be careful
Development of Neurons in Microisland Cultures Figure 12.2B shows a
microisland made with collagen on agarose. The grainy, cell-free background is
the surface of the agarose. The round cell-containing area was defined by a drop
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