Bioactive Conducting Polymers for Optimising the Neural Interface - Biointerfaces: Where Material Meets Biology - page 218

A.2.5 PC12 Neurite Outgrowth Assay
PC12 is a cell line derived from a pheochromocytoma of a rat adrenal me-
dulla. It is a non-adherent cell line which when exposed to NGF will differ-
entiate and grow neuronal processes. The cells are cultured in RPMI media
containing 10% horse serum and 5% foetal bovine serum. Although being
non-adherent, the cells will clump together and therefore require aspiration
before counting and plating. The cells can be aspirated through repeated
(

d n 3 r 4 n g | 2

70 times) drawing and expulsion through a 1000 mL pipette. Samples will
require coating in laminin in order to encourage cellular attachment; how-
ever, this may be omitted if the sample contains its own biological molecules
for cellular attachment. For the outgrowth assay cells are plated at 20 000
cells cm 2 in RPMI containing 1% horse serum and 50 ng mL 1 NGF. A two-
thirds media refresh is carried out after 48 h and the samples are analysed at
96 h using a live/dead stain. Neurite density can be determined by tracing
neurites in ImageJ using the NeuronJ plug-in.

B

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