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View additional product information for Neurobasal™ Medium - FAQs (21103049)
9 product FAQs found
Yes, the protein content in this product is high enough so filtering through a low protein binding filter as a 50X or 1X in solution should not be a problem.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
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.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
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.
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The original published formulation of Neurobasal culture medium contained 10 µM L-cysteine. However, our Neurobasal media formulation contains 260 µM L-cysteine because it was shown to improve cell survival.
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Neurospheres can be plated on laminin coated culture plates for neuron differentiation. The issue is that it is difficult to control the plating density of neurospheres. Alternatively, neurospheres can be dissociated into single cells and plate single cell suspension at a certain density such as 1-5 x 10^4 cells/cm2 onto laminin coated plates for neuron differentiation. For general neuron differentiation, Neurobasal+B27+N2 can be used. Growth factors such as BDNF and/or GDNF can be added into medium for improving survival of differentiating NSCs.
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We recommend using neural expansion medium with the following composition to expand NSCs generated using the PSC Neural Induction Medium:
- 50:50 Neurobasal medium: Advanced DMEM/F12 plus PSC Neural Induction Supplement.
PSC lines can behave differently, and we recommend performing your own validation experiments if you want to use PSC Neural Induction Medium, or you do not observe good expansion using the Neurobasal and Advanced DMEM/F12 combination.
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L-glutamate is excitotoxic as neuron cells mature. For primary hippocampal neurons and other embryonic neurons, we recommend that you add 25 µM L-glutamate to the initial plating medium. However, after day 4, L-glutamate should not be added as it is toxic to neuron cells beyond day 4.
For neuroblastomas, L-glutamate should be included in the medium for both plating and subsequent medium changes.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
As a general guideline, we recommend using Gibco B-27 Plus Supplement to culture neural stem cells, hippocampal, and other CNS neurons.
- For neuroblastomas, or post-mitotic neurons from both PNS and CNS, Gibco N-2 Supplement can be used.
- For primary glial cells (astrocytes) or tumor cell lines of astrocytic phenotype (astrocytes and gliomas), or oligodendrocytes, Gibco G-5 Supplement can be used.
Neurobasal Medium is optimized for prenatal and fetal neurons. Neurobasal-A Medium is optimized for growing postnatal and adult brain neurons. These two media differ only in osmolality.