Journal
JOURNAL OF NEUROSCIENCE METHODS
Volume 170, Issue 2, Pages 181-187Publisher
ELSEVIER
DOI: 10.1016/j.jneumeth.2008.01.009
Keywords
neuron culture; serum-free medium; synaptogenesis; electrode array; network
Categories
Funding
- NIA NIH HHS [R01 AG013435, R01 AG013435-09, R56 AG013435, AG13435] Funding Source: Medline
- NINDS NIH HHS [R01 NS052233, R01 NS052233-02, NS52233] Funding Source: Medline
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The most interesting property of neurons is their long-distance propagation of signals as spiking action potentials. Since 1993, Neurobasal/B27 (TM) has been used as a serum-free medium optimized for hippocampal neuron survival. Neurons on microelectrode arrays (MEA) were used as an assay system to increase spontaneous spike rates in media of different compositions. We find spike rates of 0.5 s(-1) (Hz) for rat embryonic hippocampal neurons cultured in Neurobasal/B27, lower than cultures in serum-based media and offering an opportunity for improvement. NbActiv4 (TM) was formulated by addition of creatine, cholesterol and estrogen to Neurobasal/B27 that synergistically produced an eightfold increase in spontaneous spike activity. The increased activity with NbActiv4 correlated with a twofold increase in immunoreactive synaptophysin bright puncta and GluR1 total puncta. Characteristic of synaptic scaling, immunoreactive GABA(A beta) puncta also increased 1.5-fold and NMDA-R1 puncta increased 1.8-fold. Neuron survival in NbActiv4 equaled that in Neurobasal/B27, but with slightly higher astroglia. Resting respiratory demand was decreased and demand capacity was increased in NbActiv4, indicating less stress and higher efficiency. These results show that NbActiv4 is an improvement to Neurobasal/B27 for cultured networks with an increased density of synapses and transmitter receptors which produces higher spontaneous spike rates in neuron networks. (C) 2008 Elsevier B.V. All rights reserved.
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