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Functional Evaluation of Biological Neurotoxins in Networked Cultures of Stem Cell-derived Central Nervous System Neurons
Published on: February 5, 2015
Excitotoxicity triggered by Neurobasal culture medium
Joshua Hogins1, Devon C Crawford, Charles F Zorumski
1Departments of Psychiatry, Washington University School of Medicine, St. Louis, Missouri, United States of America.
Plos One
|October 8, 2011
Summary
Neurobasal medium can unexpectedly kill hippocampal neurons due to high L-cysteine levels activating NMDA receptors. This excitotoxicity, preventable with D-APV, highlights a critical factor in neuronal cell culture.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- Neurobasal medium is widely used for primary neuronal cell culture.
- Unexpected toxicity was observed in postnatal hippocampal neurons upon routine medium exchange with Neurobasal.
Purpose of the Study:
- To investigate the cause of Neurobasal-induced toxicity in hippocampal neurons.
- To identify the specific component and mechanism responsible for the observed neuronal death.
Main Methods:
- Neuronal cultures were exposed to Neurobasal and Minimal Essential Medium (MEM).
- Neurobasal toxicity was assessed using D-2-Amino-5-phosphonovalerate (D-APV) and whole-cell recordings.
- L-cysteine concentration in Neurobasal was analyzed and its effects were tested.
Main Results:
- Neurobasal, but not MEM, caused significant hippocampal neuron death within minutes, preventable by D-APV.
- Neurobasal activated D-APV-sensitive currents, indicating N-methyl-D-aspartate (NMDA) receptor activation.
- Commercial Neurobasal contains high L-cysteine (260 µM), which mimics Neurobasal's toxicity and activates NMDA receptors.
Conclusions:
- High L-cysteine concentrations in commercial Neurobasal medium mediate NMDA receptor-dependent excitotoxicity in hippocampal neurons.
- The findings necessitate consideration of NMDA receptor agonist effects when using Neurobasal for neuronal development and survival studies.
- Optimizing Neurobasal formulations or adjusting usage protocols may be crucial for preventing unintended neuronal loss in research settings.
