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Gibco™ B-27™ Plus Supplement (50X)

Catalog No. A3582801
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Catalog No. A3582801 Supplier Gibco™ Supplier No. A3582801
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Description

B-27 Plus Supplement is the next-generation of the most cited neuronal cell culture product, B-27 Supplement

B-27 Plus Supplement is a serum-free supplement that provides the highest rate of in vitro survival of primary rodent and human stem cell-derived neurons when used with Neurobasal Plus Medium. B-27 Plus Supplement features an optimized formulation, upgraded manufacturing process, and more stringent quality control of raw materials and final product. These improvements enable the highest rate of neuronal cell survival, improved electrophysiological activity, and enhanced functional maturity versus other neuronal cell culture media systems.


B-27 Plus Supplement, when combined with Neurobasal Plus Medium, 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, B-27 Plus Supplement can seamlessly replace other neuronal cell culture supplements, such as classic B-27 Supplement. The B-27 Plus Neuronal Culture System is also available for purchase, which is comprised of B-27 Plus Supplement and Neurobasal Plus Medium, offering a convenient and valuable all-in-one ordering solution.

Specifications

Content And Storage Store at -5 to -20°C. Protect from light.
Cell Type Neural Cells
Classification Serum-free
Concentration 50 X
Culture Type Mammalian Cell Culture
Form Liquid
Product Type Cell Culture Supplement
Serum Level Serum-Free
Sterility Sterile
Product Line B-27
Quantity 10 mL
Shipping Condition Dry Ice
Tested For Endotoxin, Performance, Sterility
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Frequently Asked Questions (FAQs)

Can B-27 supplement be filtered?

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.

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.
Is precipitate in Human Keratinocyte Growth Supplement (Cat. No. S0015, S001K) normal?

After long-term storage at -20 degrees C, Human Keratinocyte Growth Supplement (HKGS) (Cat. Nos. S0015, S001K) may contain a small amount of precipitate, which is normal. This precipitate is formed from cold insoluble material in the BPE component of the HKGS and will not affect the performance of the product. A similar precipitate may be seen in the BPE component of the HKGS Kit.

Can I thaw B-27 Plus Supplement (Cat. No. A3582801) in a 37 degrees C water bath or incubator?

We do not recommend thawing B-27 Supplement (Cat. No. A3582801) in a water bath or incubator. We recommend thawing the B-27 Supplement at 4 degrees C overnight.

Note: Remaining B-27 Plus Supplement (50X) may be aliquoted into working volumes and stored at -20 to -5 degrees C. Do not freeze-thaw B-27 Plus Supplement (50X) more than twice. Once thawed do not leave thawed supplement at 4 degrees C for more than two weeks.

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
What is the difference between B-27 Plus Supplement and classic B-27 Supplement?

B-27 Plus Supplement is an optimized formulation of classic B-27 Supplement with a streamlined manufacturing process. No small molecules or growth factors have been added.
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.
What B-27 supplements do you offer?

We offer the following B-27 supplement products, which are developed for specific applications:

- Gibco B-27 Serum-free Supplement (Cat. No. 17504044): Complete formulation for the growth of long-term viability of neurons.
- Gibco B-27 Supplement minus AO (antioxidants) (Cat. No. 10889038): Five antioxidants are removed. The supplement is ideal in studies of oxidative damage, apoptosis, or applications in which free radical damage to neurons occurs.
- Gibco B-27 Supplement minus Insulin (Cat. No. A1895601): Insulin is removed. The supplement is ideal for studies of insulin secretion and insulin receptors.
- Gibco B-27 Supplement minus Vitamin A (Cat. No. 12587010): Vitamin A which induces neural differentiation is removed. The supplement is ideal for proliferation of stem cells.
Which supplement can I add to Neurobasal Medium to culture my neural cells?

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.

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