B-27 Supplement: The Standard for Neuronal Cell Culture

Microscopic image of neurons cultivated B-27 Plus Supplement and neurobasal media

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.

All Gibco neuronal cell culture reagents are optimized to work together as a complete media system. Learn more about Gibco neural cell culture media or primary neurons.

Table 1. Selection guide for B-27 supplements based on cell type and application.

Application	Product	Cat. No.
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.

Phase contrast microscopic image of primary cortical neurons

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

Learn more about the B-27 Plus Neuronal Culture System

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.

Graphed diameters of neural spheroids developed in B27 Supplement

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.

CTS B-27 Supplement, Xeno-Free

Commercial Cell Therapy Manufacturing Gibco B-27 Supplement (serum free) has been the trusted standard for a variety of neuronal culture applications with citations in more than 11,000 publications. As research moves toward clinical applications, Gibco CTS B-27 Supplement XenoFree enables the translation from the bench to commercial manufacturing. This Gibco Cell Therapy Systems (CTS) product offers proven serum-free supplementation for the growth, expansion, differentiation, and maintenance of pluripotent stem cell (PSC)-derived cells.

Applicable to numerous cell types such as neural stem cells, neurons, and cardiomyocytes
Three varieties available (full version, minus vitamin A, and minus insulin)
Equivalent performance to the research-use-only (RUO) Gibco B-27 Supplement XenoFree

Learn more

Citations for the use of B-27 supplement in 3D culture

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.

Cell type	Title	Citation
Brain, PSCs	Modeling Parkinson’s disease in midbrain-like organoids	Smits et al. NPJ Parkinsons Dis. 2019;5;5 (1)1–8.
Brain, PSCs	Modeling G2019S-LRRK2 sporadic Parkinson's disease in 3D midbrain organoids	Kim et al. Stem Cell Reports. 2019;12(3):518–531.
Breast	A CD146 FACS protocol enriches for luminal keratin 14/19 double positive human breast progenitors	Ísberg et al. Sci Rep. 2019; 9:14843.
Colon	Modeling the effect of prolonged ethanol exposure on global gene expression and chromatin accessibility in normal 3D colon organoids	Devall et al. PLoS ONE. 2020;15(1):e0227116.
Esophagus	Esophageal 3D organoids of MPV17-/-mouse model of mitochondrial DNA depletion show epithelial cell plasticity and telomere attrition	Guha et al. Oncotarget. 2019;10(58):6245–6259.
Fallopian tube	Chronic Chlamydia infection in human organoids increases stemness and promotes age-dependent CpG methylation	Kessler et al. Nat Commun. 2019; 0(1):1–4.
Heart	Generation of Heart Organoids Modeling Early Human Cardiac Development Under Defined Conditions	Israeli et al. (2020), BioRxiv 10.1101/2020.06.25.171611
Intestine	Extracellular vesicle release from intestinal organoids is modulated by Apc mutation and other colorectal cancer progression factors	Szvicsek et al. Cell Mol Life Sci. 2019; 76:2463–2476.
Kidney	Stem cell-derived kidney organoids: engineering the vasculature	Koning et al. Cell Mol Life Sci. 2020;77(12):2257–73.
Kidney	Organoids as a new model for improving regenerative medicine and cancer personalized therapy in renal diseases	Grassi et al. Cell Death Dis. 2019; 10(3):1–5.
Lung	Modeling of fibrotic lung disease using 3D organoids derived from human pluripotent stem cells	Strikoudis et al. Cell Rep. 2019;27(12):3709–3723.
Prostate	Prostate organoid cultures as tools to translate genotypes and mutational profiles to pharmacological responses	Pappas et al. J Vis Exp. 2019;152:e60346
Retina	Transcriptome-based molecular staging of human stem cell-derived retinal organoids uncovers accelerated photoreceptor differentiation by 9-cis retinal	Kaya et al. Mol Vis. 2019; 25:663–678.
Sweat glands	Sweat gland organoids contribute to cutaneous wound healing and sweat gland regeneration	Diao et al. Cell Death Dis. 2019;10:238.

References

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.