PMID: 11340637May 8, 2001Paper

Temperature and time interval for culture of postmortem neurons from adult rat cortex

Journal of Neuroscience Research
J VielG J Brewer

Abstract

For a model of neurological disease and ischemia, we extended recent work to culture adult postmortem rat brain neurons. Frontal cortex sections were removed from adult rats immediately following sacrifice and at different postmortem intervals and with the brain at either 22 degrees C or 4 degrees C. Brain could be stored four times longer at 4 degrees C between sacrifice and neuronal disaggregation to achieve the same 20% recovery of live cells from those plated compared to 22 degrees C. Each milligram of rat frontal cortex was estimated by the optical disector method to contain 160,000 neurons. When cells were isolated as rapidly as possible, 9% of the neurons originally present in the brain were viable. Various postmortem intervals from 2 to 24 hr resulted in a reduction from 6% to 3% of the cells originally present. After 5 days in culture, viable neurons were 23-42% of those isolated. Neuron-like cells that survived represented 40-75% of the viable cells, or 0.5-2.75% of those originally estimated to be present in the brain. Electrophysiology experiments show that cells isolated 0 and 24 hr postmortem had neuronal electrical properties, including an average resting membrane potential of -48 mV, voltage-sensitive currents, ...Continue Reading

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Citations

Aug 9, 2012·In Vitro Cellular & Developmental Biology. Animal·Mahipal SinghAnil Sharma
Jun 8, 2001·Journal of Neuroscience Methods·G J BrewerD C McManus
Nov 21, 2007·Journal of Neurochemistry·José R LopezAlexander Shtifman
Dec 30, 2014·In Vitro Cellular & Developmental Biology. Animal·Charles Okonkwo, Mahipal Singh
Jan 5, 2002·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Ronald W H VerwerDick F Swaab
Dec 16, 2016·Molecular Neurobiology·José R LopezJosé A Adams

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