B-27™ Plus Neuronal Culture System - FAQs

View additional product information for B-27™ Plus Neuronal Culture System - FAQs (A3653401)

21 product FAQs found

Which media system should I use for primary culture of P0- or P1-derived mixed hippocampal cells?

We recommend using Neurobasal Plus Medium with B-27 Plus Supplement for long-term culture of mixed hippocampal cells. For short-term culture of a pure population of hippocampal neurons, we would recommend using Neurobasal-A Medium with B-27 Supplement.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Do you offer B-27 supplements with and without insulin?

Yes, we do offer B-27 Supplement, minus insulin (Cat. Nos. A1895601 and A1895602). All our other B-27 supplements do contain insulin.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

I'm currently culturing neurons. Which media system do you recommend using in conjunction with the Poly-D-Lysine matrix?

We recommend using the B-27 Plus Neuronal Culture System when culturing neurons on Poly-D-Lysine. The B-27 Plus Neuronal Culture System is a complete media system that includes the B-27 Plus Supplement and the Neurobasal Plus Medium.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Can I use the B-27 Plus Neuronal Culture System for electrophysiological studies?

Yes, we have shown improved electrophysiological activity with the B-27 Plus Neuronal Culture System

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How do I feed my cells with the B-27 Plus Neuronal Culture System?

We recommend performing half medium exchanges every 2-3 days post-plating. Make sure that neurons are not completely exposed to air by not removing all of the medium during exchanges.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Can I thaw B-27 Supplements in a 37 degrees C water bath or incubator?

We do not recommend thawing at 37 degrees C. Best results are observed upon thawing at room temperature for approximately 2 hrs or overnight at 2-8 degrees C.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

How do I thaw B-27 Supplements?

We recommend thawing it at 4 degrees C overnight or at room temperature for 2 hours. We don't recommend thawing in a water bath or incubator.

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What is HuC/D?

HuC/D is an antibody specific to neuronal cell bodies. It can be used to quantitate neurons in culture.

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I am using the B-27 Plus Neuronal Culture System. What do I do if my neuronal cells are not attaching or are clumped?

In primary neuronal cultures, this is typically a function of uneven coating or if neurons are plated at too high of a density. In NSC differentiation experiments, this could be due to overproliferation of neural progenitor cells. Differentiating with the B-27 Plus Neuronal Culture System and CultureOne Supplement can eliminate more than 75% of neural progenitor cells and the clumps they form. Please see the CultureOne Supplement User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0016204_CultureOneSupplement_UG.pdf) for more information.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

What is the shelf-life of B-27 Plus Supplement and Neurobasal Plus Medium?

B-27 Plus Supplement can be stored for up to one year at -5 to -30 degrees C.
Neurobasal Plus Medium can be stored for up to one year at 2-8 degrees C.
Both products should be protected from light.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Can I store thawed B-27 Plus Supplement at 4 degrees C?

Yes, B-27 Plus Supplement can be stored at 4 degrees C for up to 2 weeks post-thaw.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

How long can I use Neurobasal Plus Medium supplemented with B-27 Plus Supplement?

Complete Neurobasal Plus Medium should be used within 4 weeks after supplementation with B-27 Plus Supplement.

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In which protocols would you recommend substituting B-27 Plus Supplement and Neurobasal Plus Medium for classic B-27 and neurobasal products?

We recommend using B-27 Plus Supplement and Neurobasal Plus Medium instead of the regular versions of B-27 Supplement and Neurobasal Medium products (not specialty versions like minus Insulin, minus Vitamin-A or Neurobasal-A) in protocols used for:

- Maintenance of primary rat, mouse and human PSC-derived and fetal-derived neurons
- Differentiation of human PSC-derived and fetal-derived neural stem cells (NSCs) to neurons

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Can I use B-27 Plus Neuronal Culture System to culture non-neuronal cells?

The B-27 Plus Neuronal Culture System was designed for culturing neuronal cells. We have not tested it in other cells.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

I am planning to use the B-27 Plus Neuronal Culture System. How can I control the number of glial cells in my neuronal culture?

Add CultureOne Supplement at day 0 with the B-27 Plus Neuronal Culture System to fully suppress both astrocytes and oligodendrocytes with no detrimental effect on neurons. Delaying the addition of CultureOne Supplement to later time points results in increasing levels of astrocytes. Refer to the CultureOne Supplement User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0016204_CultureOneSupplement_UG.pdf) for more information.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Do I have to use B-27 Plus Supplement with Neurobasal Plus Medium?

B-27 Plus Supplement and Neurobasal Plus Medium have been designed to work synergistically together for maximal performance. They can be used with other basal media or supplements if necessary.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

How long can I maintain my neurons in culture with the B-27 Plus Neuronal Culture System?

We have maintained primary rat cortical neurons up to 8 weeks and rat hippocampal neurons up to 4 weeks. Mouse cortical neurons were maintained for 4 weeks. Human iPSC-derived neurons that have been differentiated from neural stem cells (NSCs) for 9 days have been maintained for 5 weeks. All cells tested with the B-27 Plus System were significantly healthier than cells maintained in culture using the classic products across all time points.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Can I use the B-27 Plus Neuronal Culture System for expansion of proliferating cells, such as neural stem cells (NSCs)?

No, we do not recommend the B-27 Plus System for expanding proliferating cells. We recommend using StemPro NSC SFM, Neural Induction Medium Supplement or B-27 Supplement minus Vitamin A for growing proliferative cells, such as NSCs.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Can I re-freeze B-27 Supplements into smaller aliquots for future use?

Yes, we recommend that you aliquot when you first thaw it and thereafter subject the aliquot to no more than 2 additional freeze/thaw cycles.

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How did you upgrade the manufacturing/QC process of the B-27 Plus Supplement and Neurobasal Plus Medium?

We streamlined the manufacturing process, implemented more stringent acceptance criteria and enhanced QC assays for qualification of raw materials and release of finished product to enable better lot-to-lot consistency.

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What is the difference between Neurobasal Plus Medium and Neurobasal Medium?

Key amino acids and buffering components have been optimized in Neurobasal Plus Medium. No small molecules or growth factors have been added.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.