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Gibco™ B-27™ Plus Neuronal Culture System

Catalog No. A3653401
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Description

The B-27 Plus Neuronal Culture System is a next-generation media system that provides the highest rate of in vitro survival of primary rodent and human stem cell-derived neurons.

B-27™ Plus Neuronal Culture System is comprised of B-27 Plus Supplement (50X) and Neurobasal Plus Medium, this system is an evolution of the most cited neuronal cell culture products, B-27 Supplement and Neurobasal Medium. It features an optimized formulation, upgraded manufacturing process, and more stringent quality control of raw materials and final product. These improvements enable the highest neuronal cell survival, improved electrophysiological activity, and enhanced functional maturity versus other neuronal cell culture media systems.

The B-27 Plus Neuronal Culture System offers:

  • At least 50% higher survival rate of primary rat neurons compared to classic B-27 Supplement and Neurobasal Medium
  • Highest survival rate of primary rat, mouse, and human stem cell-derived neurons compared to other commercially available serum-free neuronal media systems
  • Improved electrophysiological activity and maturation—more consistent, stable, and synchronized spontaneous activity in primary rodent neurons after 10 days compared to BrainPhys™ Neuronal Medium
  • Accelerates neurite outgrowth and increases the number of synaptic-positive puncta

Available as a 50X, 10 mL supplement with 500 mL basal medium, the B-27 Plus Neuronal Culture System can seamlessly replace other neuronal cell culture systems, such as classic B-27 Supplement and Neurobasal Medium. The individual components of the system are also available for purchase separately.

Specifications

Cell Type Neural Cells
Classification Serum-free
Form Liquid
Product Type Neuronal Culture System
Serum Level Serum-free
Product Line B-27, Neurobasal
Quantity 1 System
Shipping Condition Ambient
Content And Storage • 10 mL B-27 Plus Supplement (50X), store at –5 to –20°C, protect from light
• 500 mL Neurobasal Plus Medium, store at 2–8°C, protect from light
Culture Type Mammalian Cell Culture

Frequently Asked Questions (FAQs)

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.
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.
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.
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
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.
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.
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.
What is HuC/D?

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

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.
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.
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.
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.
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.
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.
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.
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.

For Research Use Only. Not for use in diagnostic procedures.