Gibco™ B-27™ Plus Neuronal Culture System with Neurobasal™ PhysioPlus Medium

Catalog No.	Glucose Concentration	Without Additives
A3653401	High	None

Catalog No. A3653401 Supplier Gibco™ Supplier No. A3653401

Description

Improve the physiological relevance of neuronal cultures without sacrificing long-term health and stability. The B-27 Plus Neuronal Culture System with Neurobasal PhysioPlus Medium supports earlier, sustained neuronal activity, and exceptional cell viability to help accelerate scientific discoveries.

The B-27 Plus Neuronal Culture System combines B-27 Plus Supplement with the advanced Neurobasal PhysioPlus Medium to support healthy, functional neuronal cultures. Designed for primary and hPSC-derived neurons, the system allows a more physiologically relevant culture environment that promotes accelerated neuronal maturation, dependable neuronal activity, and sustained viability over extended culture periods. The system is suitable across neurobiology applications from functional assays including MEA and live-cell imaging to high-content imaging and high-throughput plate-reader assays.

Key Features & Benefits

Morphological and synaptic maturation

• Improves neurite outgrowth, synapse formation, neuronal morphology compared with conventional neuronal media approaches
• Accelerates neurite outgrowth and increases the number of synaptic-positive puncta

Supports earlier and sustained neuronal activity

Obtaining neurons that survive, mature, and exhibit dependable functional activity in vitro remains a major challenge in neuroscience research. The B-27 Plus Neuronal Culture System was developed to support neuronal activity that arises earlier in culture and remains stable across extended timepoints, helping researchers obtain results faster and perform longitudinal studies with greater confidence. Product performance has been verified by whole-cell patch clamp recordings and MEA experiments.

The system is optimized to support:

• Spontaneous neuronal activity
• Formation of functional neuronal networks
• Sustained electrophysiological activity over multiple weeks suitable for MEA workflows

More physiologically relevant neuronal culture conditions

Neurobasal PhysioPlus Medium is formulated to better reflect key aspects of the central nervous system (CNS) extracellular environment while maintaining robust neuronal health and performance.

• Osmolarity consistent with CNS extracellular conditions
• Reduced glucose concentrations, closer to physiological range
• Neuroactive components at relevant concentrations
• Verified electrophysiological properties of neurons in medium

Maintain healthy neurons longer in culture

The B-27 Plus Neuronal Culture System supports long-term neuron survival and health, enabling the highest survival rate of human stem cell-derived neurons and primary rodent neurons compared to other commercially available serum-free neuronal media systems. For example, 87% greater viability of rat cortical neurons was achieved with the B-27 Plus Neuronal Culture System compared to a leading competitor system, allowing your precious cell source to be seeded at lower density, seed more wells, and generate more data.

Researchers can culture neurons for extended periods to enable:

• Neuronal maturation studies
• Chronic treatment paradigms
• Disease modeling applications
• Longitudinal imaging experiments
• Repeated functional measurements

Multiple basal medium options available to meet your needs

Neurobasal PhysioPlus Medium - the recommended basal medium, formulated to better replicate the CNS extracellular environment, supporting sustained neuronal activity, improved viability, and enhanced maturity over extended culture periods.

Neurobasal PhysioPlus Medium without phenol red – for use during analysis of neurons in imaging-sensitive applications or where the presence of phenol red is undesired.

Neurobasal Plus Medium – an alternative basal medium with a low osmolality, similar to the classic Neurobasal Medium (Cat. No. 21103049)

Built on a trusted neuronal culture foundation

The original B-27 Plus Neuronal Culture System expanded upon the industry-standard Neurobasal and B-27 platform to provide enhanced neuronal robustness and culture performance. The addition of Neurobasal PhysioPlus Medium represents the next evolution of this system, helping researchers generate healthier, more functional, and more physiologically relevant neuronal cultures for advanced neuroscience research.

Specifications

• Cell Type: Neural Cells
• Classification: Serum-free
• Form: Liquid
• Product Type: Neuronal Culture System
• Serum Level: Serum-free
• Without Additives: None
• Glucose Concentration: High
• 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.