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.

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 typeTitleCitation
Brain, PSCsModeling 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 organoidsKim et al. Stem Cell Reports. 2019;12(3):518–531.
BreastA CD146 FACS protocol enriches for luminal keratin 14/19 double positive human breast progenitorsÍsberg et al. Sci Rep. 2019; 9:14843.
ColonModeling the effect of prolonged ethanol exposure on global gene expression and chromatin accessibility in normal 3D colon organoidsDevall et al. PLoS ONE. 2020;15(1):e0227116.
EsophagusEsophageal 3D organoids of MPV17-/-mouse model of mitochondrial DNA depletion show epithelial cell plasticity and telomere attritionGuha et al. Oncotarget. 2019;10(58):6245–6259.
Fallopian tubeChronic Chlamydia infection in human organoids increases stemness and promotes age-dependent CpG methylationKessler et al. Nature Commun. 2019; 0(1):1-4.
HeartGeneration of Heart Organoids Modeling Early Human Cardiac Development Under Defined ConditionsIsraeli et al. (2020), BioRxiv 10.1101/2020.06.25.171611
IntestineExtracellular vesicle release from intestinal organoids is modulated by Apc mutation and other colorectal cancer progression factorsSzvicsek et al. Cell Mol Life Sci. 2019; 76:2463–2476.
KidneyStem cell-derived kidney organoids: engineering the vasculatureKoning 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 diseasesGrassi et al. Cell Death Dis. 2019; 10(3):1-5.
LungModeling of fibrotic lung disease using 3D organoids derived from human pluripotent stem cellsStrikoudis et al. Cell Reports. 2019;27(12):3709–3723.
ProstateProstate organoid cultures as tools to translate genotypes and mutational profiles to pharmacological responsesPappas et al. J Vis. 2019;152:e60346
RetinaTranscriptome-based molecular staging of human stem cell-derived retinal organoids uncovers accelerated photoreceptor differentiation by 9-cis retinalKaya et al. Mol Vis. 2019; 25:663–678.
Sweat glandsSweat gland organoids contribute to cutaneous wound healing and sweat gland regenerationDiao 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

ApplicationProduct nameCat. No.
Maintenance/maturation of prenatal/fetal primary neurons
B-27 Plus SupplementA3582801

Neural stem / progenitor cells

ApplicationProduct nameCat. No.
Proliferation of neural stem cellsB-27 Supplement, minus vitamin A12587010
Differentiation and maintenance/maturation of stem cell–derived neuronsB-27 Supplement
Maintenance/maturation of stem cell–derived neuronsB-27 Plus SupplementA3582801

Specialty applications

ApplicationProduct nameCat. No.
Studies of oxidative stress/damage, apoptosis, or where free radical damage to neurons occursB-27 Supplement minus AO (antioxidants)10889038
Studies of insulin secretion or insulin receptorsB-27 Supplement minus InsulinA1895601
Electrophysiology studiesB-27 Plus Neuronal Culture SystemA3653401

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

Primary cellsBrainstem, cerebellar, cortical, dopaminergic, DRG, enteric, GABAergic, glutamatergic, hippocampal, motor, neural progenitor, pancreatic, parasympathetic, retinal, septal, striatal, substantia nigral, sympathetic ganglia
Stem cellsInduced pluripotent stem cell, neural precursor, neural stem cell, neuroepithelial, cardiomyocytes
Glial cellsAstrocyte, microglia, oligodendrocytes, radial glia, satellite, Schwann cells
Disease modelsAlzheimer’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