Can Neurobasal Medium alone or Neurobasal Medium supplemented with B-27 supplement be frozen to extend product shelf life?
No, it is not recommended to freeze media due to the potential of precipitates forming upon thaw. Inorganic salts and amino acids in the formulation may come out of solution when exposed to temperature fluctuations. These precipitates will not go back into solution easily once formed.
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What supplements should I use with Neurobasal or Neurobasal-A medium?
Neurobasal and Neurobasal-A media (for postnatal and adult neurons) allow for long-term maintenance of neuronal cells without the need for an astrocyte feeder layer. These media should be supplemented with either serum or a serum-free supplement, plus 0.5mM L-glutamine. B-27 supplement is a serum-free supplement that comes as a 50X concentrate in a 10ml volume. This is enough supplement for 500ml of media. Fetal, postnatal, and adult neural cultures can be grown in the appropriate Neurobasal medium supplemented with B-27 supplement .
We also have two other supplements. One is called G-5 and is for growth and expression of glial cells (normal and tumor) of astrocytic phenotype. This comes in a 1ml size, at a 100X concentration. The other supplement is called N-2 and is for growth and expression of post-mitotic neurons and tumor cells of neuronal phenotype. This comes in a 5ml size, at a 100X concentration.
For more information on Neurobasal media, search "Neurobasal" from our website home page.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
Can you provide references for growing adult neurons?
Instructions for growing adult neurons can be found in the Neurobasal-A Medium product insert. You can also refer to the following publication for growing adult neurons:
• Isolation and Culture of Adult Rat Hippocampal Neurons by Brewer, G.J., J. Neuroscience Methods 71 (1997), 145-158.
Should I use Neurobasal-A Medium with B27 and b-FGF for postnatal neurons?
We do not know at what age postnatal neurons require FGF and a higher osmolality medium, or how this could vary based on the species from which the neural tissue is isolated. Without this knowledge, we generally recommend switching to the Neurobasal-A Medium for isolates from animals above 1 week of age. You will need to determine if the Neurobasal-A Medium is optimal for neurons you have from one day old rats.
Further, the addition of bFGF can increase the viability of neurons of any age. Therefore, caution needs to be made in impure neuronal cell culture as bFGF can increase the proliferation of non-neuronal cells. We recommend a pilot experiment to test the impure culture in the presence and absence of bFGF.
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What should I add to Neurobasal-A Medium to grow adult neurons?
We recommend adding B27 and b-FGF to Neurobasal-A Medium to grow adult neurons.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
