PMID- 25756909
OWN - NLM
STAT- MEDLINE
DCOM- 20151214
LR  - 20150326
IS  - 1473-558X (Electronic)
IS  - 0959-4965 (Linking)
VI  - 26
IP  - 6
DP  - 2015 Apr 15
TI  - Neurobasal medium toxicity on mature cortical neurons.
PG  - 320-4
LID - 10.1097/WNR.0000000000000343 [doi]
AB  - Neurobasal medium (NBM) is a widely used medium for neuronal cultures, originally 
      formulated to support survival of rat hippocampal neurons, but then optimized for 
      several other neuronal subtypes. In the present study, the toxic effect of NBM on 
      long-term cortical neuron cultures has been reported and investigated. A 
      significant neuronal cell loss was observed 24 h after the total medium change 
      performed at days in vitro 10. The neurotoxic effect was specifically because of 
      NBM-A, a commercially derived modification of classic NBM, as neurons exposed to 
      minimum essential medium for 24 h did not show the same mortality rate. We showed 
      that the toxic effect was mediated by the N-methyl-D-aspartate receptor (NMDAr) 
      as its inactivation partly prevented NBM-induced neuronal loss, and the addition 
      of NMDAr activators, such as L-cysteine or glycine to minimum essential medium, 
      reproduced the same toxicity rate observed in NBM. Besides the toxicity 
      associated with NMDAr activation, the decreased antioxidative defenses also 
      worsen (because of glutathione depletion) neuronal death, thus amplifying the 
      effect of excitotoxic amino acids. Indeed, glutathione supplementation by the 
      addition of its precursor N-acetyl-cysteine resulted in an increase in neuronal 
      survival that partially prevented NBM-A toxicity. These results evidenced, on the 
      one hand, the unsuitability of NBM-A for long-term neuronal culture, and on the 
      other, they highlight the importance of selection of more suitable culture 
      conditions.
FAU - Maggioni, Daniele
AU  - Maggioni D
AD  - Department of Surgery and Translational Medicine, University of Milan-Bicocca, 
      Monza, Italy.
FAU - Monfrini, Marianna
AU  - Monfrini M
FAU - Ravasi, Maddalena
AU  - Ravasi M
FAU - Tredici, Giovanni
AU  - Tredici G
FAU - Scuteri, Arianna
AU  - Scuteri A
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Neuroreport
JT  - Neuroreport
JID - 9100935
RN  - 0 (Culture Media, Conditioned)
RN  - 0 (Receptors, N-Methyl-D-Aspartate)
RN  - K848JZ4886 (Cysteine)
RN  - TE7660XO1C (Glycine)
SB  - IM
MH  - Animals
MH  - Cell Survival/drug effects
MH  - Cerebral Cortex/*drug effects
MH  - Culture Media, Conditioned/*toxicity
MH  - Cysteine/analogs & derivatives/toxicity
MH  - Glycine/analogs & derivatives/toxicity
MH  - Neurons/*drug effects
MH  - Primary Cell Culture/*methods
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Receptors, N-Methyl-D-Aspartate/metabolism
EDAT- 2015/03/11 06:00
MHDA- 2015/12/15 06:00
CRDT- 2015/03/11 06:00
PHST- 2015/03/11 06:00 [entrez]
PHST- 2015/03/11 06:00 [pubmed]
PHST- 2015/12/15 06:00 [medline]
AID - 10.1097/WNR.0000000000000343 [doi]
PST - ppublish
SO  - Neuroreport. 2015 Apr 15;26(6):320-4. doi: 10.1097/WNR.0000000000000343.