Montage image of fluorescently labeled neurons in B27 Supplement

The standard for neuronal cell culture

More than 25 years ago, the first commercially available, serum-free supplement for neural cell culture, Gibco B-27 Supplement (50X) was developed and with Gibco Neurobasal Medium has set the standard in neuronal cell culture reagents. Originally designed for culturing primary neurons, the classic B-27 Supplement and nutritive variants have been widely adapted 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.

The most-referenced neuronal cell culture supplement just got better

To further meet the growing needs of researchers, an optimized formulation of the classic B-27 Supplement was developed in 2018 for improved long-term neuronal survival, functionality and maturation of primary rodent neurons and stem cell-derived neurons. See below section: Superior survival, maturation, and neural network activity and learn more about B-27 Plus Neuronal Culture System.

The history behind the B-27 media system

For over 25 years, researchers have been publishing their experiments using Gibco neural cell culture reagents. Since its introduction, the publication history of B-27 supplements demonstrates that researchers are expanding the use of these supplements far beyond their original applications.

Interview with biomedical engineering researcher, Dr. Greg Brewer

Superior survival, maturation, and neural network activity

Maintaining healthy long-term cultures (3 weeks and beyond) of primary rodent neurons can be challenging, as these cells are quite sensitive and tend to undergo progressive cell death during culture.

Phase contrast microscopic image of primary cortical neurons
Figure 1. Primary rat cortical neurons cultured in the B-27 Plus Supplement for 3 weeks demonstrates extensive neurite networks (left image) and high expression of synapsin (right image).

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

Advancing 3D neural cell culture

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 supplements.

Scientists are discovering that B-27 supplements benefit more than just neural cell types in 2D adherent cultures. See Table 1 Citations for the use of B-27 Supplement in 3D culture.

Figure 2. Neural spheroid maturation. (A) Phase contrast images of neural spheroids at 21 days cultured in “classic” B-27 or B-27 Plus (not shown) supplement to yield spheroids of similar size and shape. (B) Gene expression analysis using TaqMan PCR assay shows increased expression level of genes related to functional synapses and neurotransmitter trafficking with the addition of CultureOne Supplement (Cat. No A3320201).

Graphed diameters of neural spheroids developed in B27 Supplement

Figure 3. Neural stem cells (NSCs) produce spheroids in B-27 Supplement. NSCs generated from Gibco Human Episomal iPSC Line (Cat No.  A18945), or H9 hESC, were cultured in maturation medium consisting of either B-27 Supplement with Neurobasal Medium (Cat No. 21103049) or in B-27 Plus Neuronal Culture System. Cells seeded in Gibco Nunclon Sphera 96-Well Microplates (Cat No. 174925) across a range of densities formed single neural spheroids within 24 hours.

Table 1. Citations for the use of B-27 Supplement in 3D culture.

Cell type Title Citation
Brain, PSCs Modeling Parkinson’s disease in midbrain-like organoids Smits et al. NPJ Parkinson’s Dis. 2019;5;5 (1)1-8.
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. Nature 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.
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 Reports. 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. 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.

Which B-27 supplement is right for you?

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. Need a complementary media or cells? Browse our neural cell culture product guide.

Standard applications / Primary neurons

Application Product name Cat. No.
Maintenance/maturation of prenatal/fetal primary neurons
B-27 Plus Supplement A3582801

Neural stem / progenitor cells

Application Product name Cat. No.
Proliferation of neural stem cells B-27 Supplement, minus 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

Specialty applications

Application Product name Cat. No.
Studies of oxidative stress/damage, apoptosis, or where free radical damage to neurons occurs B-27 Supplement minus AO (antioxidants) 10889038
Studies of insulin secretion or insulin receptors B-27 Supplement minus Insulin A1895601
Electrophysiology studies B-27 Plus Neuronal Culture System A3653401

Table 2. Cell types and disease models cited in publications using Gibco neuronal cell culture reagents.

Primary cells Brainstem, cerebellar, cortical, dopaminergic, DRG, enteric, GABAergic, glutamatergic, hippocampal, motor, neural progenitor, pancreatic, parasympathetic, retinal, septal, striatal, substantia nigral, sympathetic ganglia
Stem cells Induced pluripotent stem cell, neural precursor, neural stem cell, neuroepithelial, cardiomyocytes
Glial cells Astrocyte, microglia, oligodendrocytes, radial glia, satellite, Schwann cells
Disease models Alzheimer’s disease, amyotrophic lateral sclerosis (ALS), brain tumor, cerebral palsy, epilepsy, glioma, Huntington’s disease, multiple sclerosis, Parkinson’s disease, schizophrenia, spinal cord injury, stroke