More than 25 years ago, Dr. Greg Brewer designed and optimized B-27 neuronal culture supplement to improve survival of primary neurons in culture. Soon after, Gibco B-27 Supplement became the first commercially available serum-free neuronal cell culture supplement. The classic B-27 Supplement and nutritive variants have been widely adopted and successfully used in many other applications and cell types, including growth and maintenance of neural stem cells and PSC-derived neurons, 3D cell culture, and disease modeling.
What does B-27 neuronal culture supplement contain?
B-27 supplement is a defined yet complex mixture of antioxidant enzymes, proteins, vitamins, and fatty acids that are combined in optimized ratios to support neuronal survival in culture. The original serum-free neuronal culture supplement formula developed by Dr. Gregory Brewer and colleagues is described in Brewer et al., J Neuroscience Res 35: 567-576, 1993 and Brewer and Cotman, Brain Res 494: 65-74, 1989 [1,2].
Thermo Fisher Scientific provides convenient, lot-tested B-27 supplements based on Dr. Brewer’s formulation to support neuroscience researchers around the globe. Variations on B-27 neuronal culture supplement include formulations without insulin and B-27 without Vitamin A.
Video: Meet a pioneer of primary neuronal cell culture
Interview with biomedical engineering researcher, Dr. Greg Brewer
B-27 supplement selection guide
Choose the relevant application below to locate and order the recommended B-27 supplement for your research.
Table 1. Selection guide for B-27 supplements based on cell type and application.
|Maintenance/maturation of pre-natal/fetal primary neurons||B-27 Plus supplement||A3582801|
|Maintenance/maturation of post-natal and adult brain neurons||B-27 Plus supplement||A3582801|
|Proliferation of neural stem cells||B-27 supplement without vitamin A||12587010|
|Differentiation and maintenance/maturation of stem cell-derived neurons||B-27 supplement||17504044|
|Maintenance/maturation of stem cell-derived neurons||B-27 Plus supplement||A3582801|
|Studies of oxidative stress/damage, apoptosis, or where free radical damage to neurons occurs||B-27 supplement without AO (antioxidants)||10889038|
|Studies of insulin secretion or insulin receptors||B-27 supplement without insulin||A1895601|
|Electrophysiology studies||B-27 Plus Neuronal Culture System||A3653401|
Promote neuron survival, maturation, and neural network activity
Maintaining healthy long-term cultures (three weeks and beyond) of primary neurons can be challenging, as these cells are quite sensitive and tend to undergo progressive cell death during culture. B-27 Plus supplement promotes survival and maturation of primary neurons in culture.
Figure 1. Primary rat cortical neurons cultured in the B-27 Plus supplement for three weeks demonstrates extensive neurite networks (left) and high expression of synapsin (right).
Make the switch to the next-generation B-27 supplement
The desire for more reliable and biologically relevant models has increased, so too has the necessity for a next-generation media system that can maintain and mature optimal densities of functional neurons over longer periods of time in vitro.
The next-generation Gibco B-27 Plus Neuronal Culture System, composed of B-27 Plus supplement and Neurobasal Plus Medium, improves upon the classic culture environment through raw material and manufacturing upgrades and minor formulation modifications. Together, these small changes yield big results in the maintenance and maturation of primary neurons to offer the following improved benefits:
- Increased neuronal survival by more than 50% vs competitors
- Improved neurite outgrowth
- Improved electrophysiological activity and maturation
Advancing 3D neural cell culture with B-27 supplement
Complex 3D models are becoming a favored system to study embryonic development and disease to more faithfully recapitulate in vivo neural architectures and physiology than traditional 2D cultures. A plethora of applications, spanning a wide range of cell types have benefited from B-27 neuronal culture supplements.
Figure 2. Neural stem cells (NSCs) produce spheroids in B-27 supplement. NSCs generated from Gibco Human Episomal iPSC Line, or H9 hESC, were cultured in maturation medium consisting of either B-27 supplement with Neurobasal Medium or in B-27 Plus Neuronal Culture System. Cells seeded in Gibco Nunclon Sphera 96-Well Microplates across a range of densities formed single neural spheroids within 24 hours.
Scientists are discovering that B-27 supplements can support three-dimensional cultures of neural and other cell types, enabling a wide variety of experiments using 3D cell cultures.
Table 2. Citations for the use of B-27 supplement in 3D organoid or spheroid culture.
- Brewer, G.J.; Torricelli, J.R.; Evege, E.K.; Price, P.J. Optimized survival of hippocampal neurons in B27-supplemented Neurobasal, a new serum-free medium combination.Neurosci Res. 1993 Aug 1;35(5):567-76. doi: 10.1002/jnr.490350513.
- Brewer, G.J.; Cotman, C.W. Survival and growth of hippocampal neurons in defined medium at low density: advantages of a sandwich culture technique or low oxygen. Brain Res. 1989 Aug 7;494(1):65-74. doi: 10.1016/0006-8993(89)90144-3.
Resources and support for B-27 Supplement
- Axonal transport enables neuron-to-neuron propagation of human coronavirus OC43
- Directed differentiation of human pluripotent stem cells into intestinal tissue in vitro
- Infection of bat and human intestinal organoids by SARS-CoV-2
- Recapitulation of SARS-CoV-2 infection and cholangiocyte damage with human liver organoids
- In vitro generation of human pluripotent stem cell derived lung organoids
- A human pluripotent stem cell-based platform to study SARS-CoV-2 tropism and model virus infection in human cells and organoids
For Research Use Only. Not for use in diagnostic procedures.