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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On the culture preparation day, the merosin solution is aspirated, and plates are
incubated in medium composed of Dulbecco's modified Eagle's medium [D-MEM,
high glucose, with sodium pyruvate, without glutamine (Irvine Scientific, Irvine, ...
At present, we routinely plate the cells in Dulbecco's modified Eagle's medium (D
-MEM) basal medium containing 4% fetal calf serum and B-27 supplements (
Gibco, Gaithersburg, MD). Our first experiments showed that MHb neurons did
19. Add 300 ul of soybean trypsin inhibitor and DNase solution for 5 min.
Centrifuge as described. Remove supernatant and add 500 pil of D-MEM-BS.
Triturate into a cell suspension, beginning with gentle passage through a 5-ml
pipette and ...
In all cases, barring the generation of type-2 astrocytes, cells are grown in D-
MEM-BS on PLL-coated glass coverslips, slide flasks, tissue-culture flasks, and
the like. To generate type-2 astrocytes, in contrast, cells simply are grown in D-
Table 19.1 BASE MEDIUM NERVE GROWTH SERUM FACTOR SUPPLEMENTS
Mainlémantë fe?ds CH5, CBS E-MEM D10 D-MEM DM Ham's F-12 Fibroblast
elimination CF E-MEM E2F E-MEM Myelinating feeds MF (Schwann cells) E-
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