B27 supplement

Manufactured by Thermo Fisher Scientific

Sourced in United States, United Kingdom, Germany, Japan, Canada, China, Italy, France, Switzerland, Spain, Israel, Australia, Austria, Poland, Denmark, Palestine, State of

B27 supplement is a serum-free and animal component-free cell culture supplement developed by Thermo Fisher Scientific. It is designed to promote the growth and survival of diverse cell types, including neurons, embryonic stem cells, and other sensitive cell lines. The core function of B27 supplement is to provide a defined, optimized combination of vitamins, antioxidants, and other essential components to support cell culture applications.

Automatically generated - may contain errors

Lab products found in correlation

3 208 protocols using b27 supplement

Neuronal Differentiation Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Expansion medium (day 0-4): DMEM/F-12 GlutaMAX, (GIBCO), 2% B27 supplement (GIBCO), 1% N2 supplement (GIBCO), 1% Pen/Strep (GIBCO), 20 ng/ml EGF, 20 ng/ml bFGF.
Induction medium I (days 5-6): DMEM/F-12 GlutaMAX, (GIBCO), 2% B27 supplement (GIBCO), 1% N2 supplement (GIBCO), 1% Pen/Strep (GIBCO), 10 ng/ml EGF, 10 ng/ml bFGF.
Induction medium II (days 7-14): DMEM/F-12 GlutaMAX, (GIBCO), 2% B27 supplement (GIBCO), 1% N2 supplement (GIBCO), 1% Pen/Strep (GIBCO), 5 ng/ml bFGF.
Differentiation medium (days 15-42): Neurobasal medium (GIBCO), 2% B27 supplement (GIBCO), 1% Pen/Strep (GIBCO), 0,25% L-glutamine, 50 ng/ml BDNF.

Ciarpella F., Zamfir R.G., Campanelli A., Ren E., Pedrotti G., Bottani E., Borioli A., Caron D., Di Chio M., Dolci S., Ahtiainen A., Malpeli G., Malerba G., Bardoni R., Fumagalli G., Hyttinen J., Bifari F., Palazzolo G., Panuccio G., Curia G, & Decimo I. (2021). Murine cerebral organoids develop network of functional neurons and hippocampal brain region identity. iScience, 24(12), 103438.

+ Open protocol

+ Expand

Efficient Motor Neuron Differentiation

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Motor neurons were generated using a pre-established protocol [37 (link)]. First, neuroepithelial cells were generated by culturing iPSCs in N2B27 media (50% Neurobasal™, 50% DMEM:F12, 0.5× N2 supplement, 0.5× B27™ supplement, and 1× GlutaMAX™ (all Thermo Fisher)) supplemented with 3 μM CHIR99021 (Tocris, Bristol, UK), 2 μM dorsomorphin (Tocris), and 2 μM SB431542 (Tocris) for four days. Neuroepithelial cells were expanded and differentiated into motor neuron progenitors with N2B27 media supplemented with 0.1 μM retinoic acid (RA) (Sigma-Aldrich) and 0.5 μM Purmorphamine (Tocris) for another two days. On day six, CHIR99021, dorsomorphin and SB431542 were withdrawn, and cells were cultured with RA and dorsomorphin for an additional six days. Media was transitioned to maturation media (BrainPhys™ Neuronal Culture Media (Stem Cells technologies, Vancouver, BC, Canada), 0.5× N2 supplement (Thermo Fisher), 0.5× B27™ supplement (Thermo Fisher), 10 ng/mL BDNF (PerpoTech), and 10 ng/mL GDNF (Peprotech)) including 0.1 μM Compound E (Calbiochem, San Diego, CA, USA) for the first three days to induce terminal differentiation via Notch inhibition.

Hedges E.C., Cocks G., Shaw C.E, & Nishimura A.L. (2023). Generation of an Open-Access Patient-Derived iPSC Biobank for Amyotrophic Lateral Sclerosis Disease Modelling. Genes, 14(5), 1108.

+ Open protocol

+ Expand

Primary Cortical Neuron Culture Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary cortical neuron cultures were prepared from cerebral cortices of E15 mouse pups. Timed pregnant females were anesthetized with isoflurane and sacrificed by decapitation. Pups were removed and decapitated to allow for removal of the brains. Following removal of the meninges, cortices were incubated on ice in Accutase® Cell Detachment Solution (Innovative Cell Technologies, San Diego, CA) and dissociated step-wise using 1,000 -µL and 200 -µL sized pipette tips. Cells were counted and diluted in plating medium consisting of Neurobasal® medium (Thermo Fisher Scientific, Waltham, MA; Formulation detailed in Supplementary Table 1) supplemented with glutamate (25 µM, Sigma-Aldrich, St. Louis, MO), glutamine (0.5 mM, Thermo Fisher Scientific, Waltham, MA), Antibiotic-Antimycotic (Thermo Fisher Scientific, Waltham, MA), and B-27® supplement (Thermo Fisher Scientific, Waltham, MA). Cells were then plated at a density of 3000 cells/cm² on poly-d-lysine (1 mg/mL; Sigma-Aldrich, St. Louis, MO)-coated glass coverslips (12 mm) in 24-well plates. Following cell attachment, wells were filled with the medium described above, in which the B-27® supplement was replaced with B-27® supplement minus antioxidants (Thermo Fisher Scientific, Waltham, MA). All experiments were performed starting at 12 days in vitro (DIV).

Clark K.C., Sword B.A, & Dupree J.L. (2017). Oxidative Stress Induces Disruption of the Axon Initial Segment. ASN NEURO, 9(6), 1759091417745426.

+ Open protocol

+ Expand

Isolating Cerebellar Granule Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols

P7 cerebella were dissected and dispersed using the Neural Tissue Dissociation Kit P (Miltenyi) as recommended. The tissue was titurated into a single-cell suspension by using fire-polished glass Pasteur pipettes. CGNs were isolated by centrifugation over a discontinuous Percoll gradient, and the higher-density fraction, which contained 95% CGNs and 5% glia, was collected. CGNs were nucleofected with plasmid DNA by using the optimized Amaxa Mouse Neuron Nucleofector Kit (Lonza) with the O-005 program. The cells were recovered for 5 min then plated over poly-L-ornithine-, Matrigel-, laminin-, or vitronectin-coated 16-well slides or glass-bottom dishes (EMS) in Neurobasal medium supplemented with 0.5% glucose, 0.4 mg/mL of tissue culture-grade bovine serum albumin (Sigma), 2 mM l-glutamine, 50 U/mL penicillin–streptomycin, and 1× B27 supplement (ThermoFisher). When specified, insulin-free B27 supplement (ThermoFisher) was used in place of the standard B27 supplement.

Ong T., Trivedi N., Wakefield R., Frase S, & Solecki D.J. (2020). Siah2 integrates mitogenic and extracellular matrix signals linking neuronal progenitor ciliogenesis with germinal zone occupancy. Nature Communications, 11, 5312.

+ Open protocol

+ Expand

Neuronal Differentiation Media Formulations

Check if the same lab product or an alternative is used in the 5 most similar protocols

N2B27 medium is composed of DMEM/F12 supplemented with modified N2 supplement and Neurobasal medium supplemented with B27 supplement minus vitamin A, in a 1:1 ratio and 50 μM β-mercaptoethanol (Thermo Scientific). FEB medium is N2B27 medium supplemented with 10 ng/mL FGF-2 (Peprotech), 10 ng/mL EGF (R&D Technologies), and 20 ng/mL BDNF (Peprotech). SFA medium is N2B27 medium supplemented with 100 ng/mL FGF-8 (Peprotech), 200 ng/mL sonic hedgehog (Peprotech), and 100 μM ascorbic acid 2-phosphate (Sigma). BGAA medium is N2B27 medium supplemented with 20 ng/mL BDNF, 10 ng/mL GDNF (Peprotech), 500 μM dibutyryl cyclic AMP (Sigma), 100 μM ascorbic acid 2-phosphate, 100 units/mL of penicillin, 100 μg/mL of streptomycin (Pen-Strep, Thermo Scientific), and 2 μg/mL Laminin (Roche).
Cyno NPC differentiation medium is composed of Neurobasal medium supplemented with B27 supplement (Thermo Scientific) and GlutaMAX supplement (Thermo Scientific), 20 ng/mL BDNF, 10 ng/mL GDNF, 100 μM ascorbic acid 2-phosphate, 100 units/mL of penicillin, 100 μg/mL of streptomycin, and 2 μg/mL Laminin.

Easton A., Jensen M.L., Wang C., Hagedorn P.H., Li Y., Weed M., Meredith J.E., Guss V., Jones K., Gill M., Krause C., Brown J.M., Hunihan L., Natale J., Fernandes A., Lu Y., Polino J., Bookbinder M., Cadelina G., Benitex Y., Sane R., Morrison J., Drexler D., Mercer S.E., Bon C., Pandya N.J., Jagasia R., Ou Yang T.H., Distler T., Grüninger F., Meldgaard M., Terrigno M., Macor J.E., Albright C.F., Loy J., Hoeg A.M., Olson R.E, & Cacace A.M. (2022). Identification and characterization of a MAPT-targeting locked nucleic acid antisense oligonucleotide therapeutic for tauopathies. Molecular Therapy. Nucleic Acids, 29, 625-642.

+ Open protocol

+ Expand

Neuronal Differentiation of iPSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

iPSCs were cultured in Matrigel‐coated tissue culture dish with TesR‐E8 (STEMCELL Technologies). When 15%‐25% confluence was reached, iPSC culture media was replaced with neural induction (NIM1) medium 1 (DMEM/F12 supplemented with L‐glutamine [2 mM], N2, B27 supplement, bovine serum albumin [1 mg/mL, ThermoFisher], NEAA [10 mM; ThermoFisher], SB431542 [10 μM; STEMCELL Technologies], noggin [200 ng/mL, GenScript], and laminin [1 μg/mL, Sigma‐Aldrich]). Afterward, medium was replaced with fresh prewarmed NIM1 after every 48 hours. After 1 week, when cells were reaching maximum confluence, NIM1 was switched to NIM2 (DMEM/F12 supplemented with L‐glutamine [2 mM], N2, B27 supplement, bovine serum albumin [1 mg/mL, ThermoFisher], NEAA [10 mM; ThermoFisher], and laminin [1 μg/mL, Sigma‐Aldrich]). Cells were cultured in NIM2 for 5 days, with daily media change. After 5 days, cells were dissociated using Gentle Cell Dissociation Reagent (STEMCELL Technologies) and cultured in low attachment dishes for 2 days with STEMDiff neural progenitor medium (STEMCELL Technologies). Formed aggregates were then plated onto polyornithine‐/laminin‐coated tissue culture dish for mNPCs expansion and cultured with the same media.

Jefri M., Bell S., Peng H., Hettige N., Maussion G., Soubannier V., Wu H., Silveira H., Theroux J., Moquin L., Zhang X., Aouabed Z., Krishnan J., O'Leary L.A., Antonyan L., Zhang Y., McCarty V., Mechawar N., Gratton A., Schuppert A., Durcan T.M., Fon E.A, & Ernst C. (2020). Stimulation of L‐type calcium channels increases tyrosine hydroxylase and dopamine in ventral midbrain cells induced from somatic cells. Stem Cells Translational Medicine, 9(6), 697-712.

+ Open protocol

+ Expand

Cardiac Differentiation of hESCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

At day 0 of cardiac differentiation, hESCs were incubated with basal medium comprising RPMI 1640 (Invitrogen) plus B27 supplement minus insulin (A1895601; Thermo Fisher, Waltham, MA, USA). On days 0–1, 6 μM CHIR-99021 (HY-10182; MCE, Monmouth Junction, NJ, USA) was added to the medium. On days 3–5, 2 μM wnt-C59 (HY-15659; MCE) was added to the medium. On day 7 of differentiation, the medium was changed to RPMI 1640 plus B27 supplement (17504044; Thermo Fisher). The medium was changed every 48 h. Beating cardiomyocytes appeared starting on day 7 of differentiation.

Han Z., Xu Z., Yu Y., Cao Y., Bao Z., Gao X., Ye D., Yan G., Gong R., Xu J., Zhang L., Ma W., Wang X., Yang F., Lei H., Tian Y., Hu S., Bamba D., Li Y., Li D., Li C., Wang N., Zhang Y., Pan Z., Yang B, & Cai B. (2021). ALKBH5-mediated m6A mRNA methylation governs human embryonic stem cell cardiac commitment. Molecular Therapy. Nucleic Acids, 26, 22-33.

+ Open protocol

+ Expand

Differentiation of human iPSCs into Cardiomyocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

To differentiate human iPSCs into cardiomyocytes, cells were cultured in mTeSR^™1 media until they reached 80 to 90% confluency, and cells were cultured in CDM3-C media, consisting of RPMI 1640 (Life Technologies, 11875), 500 μg/mL Oryza sativa-derived recombinant human albumin (A0237, Sigma-Aldrich), and 213 μg/mL L-ascorbic acid 2-phosphate supplemented with 10 μM CHIR-99021 (Selleckchem, S2924) for two days. Cells were then cultured in CDM3-C media, supplemented with 2 μM WNT-C59 (Selleckchem, S7037) for 2 days. Cells were cultured in BASAL media (RPMI-1640 with B27 Supplement (Thermo Fisher Scientific, 17504044)) for 6 days, and media was changed every 2 days. Cardiomyocytes were selected by culturing in SELECTIVE media (RPMI-1640, no glucose (Gibco, 11879–020) with B27 Supplement (Thermo Fisher Scientific, 17504044)) for 6 days. Then, purified cardiomyocytes were dissociated using TrypLE Express Enzyme (Gibco, 12604021) and replated at 1×10⁵ cells per well in a 6-well dish.

Gan P., Wang Z., Morales M.G., Zhang Y., Bassel-Duby R., Liu N, & Olson E.N. (2022). RBPMS is an RNA-binding protein that mediates cardiomyocyte binucleation and cardiovascular development. Developmental cell, 57(8), 959-973.e7.

+ Open protocol

+ Expand

Neurosphere Dissociation and Culture

Check if the same lab product or an alternative is used in the 5 most similar protocols

Neurospheres were dissociated into single cells via incubation with TrypLE Select (Thermo Fisher Scientific) at 37 °C for 5 min and seeded on a Matrigel (Corning, Corning, NY, USA)-coated plate at a density of 7.5 × 10⁴ cells/cm² in DMEM/F12 supplemented with 2% B-27 Supplement (Thermo Fisher Scientific) plus 1% fetal bovine serum (FBS; GE healthcare, Chicago, IL, USA) (the 1% serum medium), or Neurobasal Medium (Thermo Fisher Scientific) supplemented with 2% B-27 Supplement plus 1% GlutaMAX (Thermo Fisher Scientific) (the serum-free medium).

Kanemura Y., Yamamoto A., Katsuma A., Fukusumi H., Shofuda T., Kanematsu D., Handa Y., Sumida M., Yoshioka E., Mine Y., Yamaguchi R., Okada M., Igarashi M., Sekino Y., Shirao T., Nakamura M, & Okano H. (2024). Human-Induced Pluripotent Stem Cell-Derived Neural Progenitor Cells Showed Neuronal Differentiation, Neurite Extension, and Formation of Synaptic Structures in Rodent Ischemic Stroke Brains. Cells, 13(8), 671.

+ Open protocol

+ Expand

Astrocyte Culture and Viral Transduction

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary astrocyte cell cultures were prepared according to a previously described protocol 31 with slight modifications: Hippocampi were isolated from brains of neonatal mice between P1-3 and cells were seeded after dissociation at a density of 5x10 4 cells per 12 mm glass coverslip for microscopy in 500 µl plating medium (49 ml MEM, 1 ml B-27 supplement, 500 µl sodium pyruvate, 500 µl L-Glutamine, 50 µl Penicillin-Streptomycin; all Thermo Fisher Scientific Inc., Waltham, USA). On DIV3 the entire plating medium was replaced with 1 ml maintenance medium (49 ml Neurobasal-A, 1 ml B-27 supplement, 500 µl L-Glutamine, 50 µl Penicillin-Streptomycin; all Thermo Fisher Scientific Inc., Waltham, USA). On DIV11, ½ of the medium was exchanged with prewarmed maintenance medium prior to infection of the cells with of 0.1 µl AAV-mGFAP-GCaMP6s (3.7 x 10 9 vg/µl) and AAV-mGFAP-tdTomato (1 x 10 7 vg/µl). Astrocytes were maintained at 37 °C in a humidified incubator in a 5% CO2 atmosphere used for experiments between DIV14-17. Cells were transferred to a prewarmed recording chamber for microscopy and kept in a balanced salt solution (BSS), which was adjusted to pH 7.4 and 290 mOsm with glucose, containing 115 mM NaCl, 5.4 mM KCl, 1 mM MgCl2, 2 mM CaCl2 and 20 mM HEPES.

Müller F.E., Cherkas V., Stopper G., Caudal L.C., Stopper L., Kirchhoff F., Henneberger C., Ponimaskin E., & Zeug A. (2020). Deciphering spatio-temporal fluorescence changes using multi-threshold event detection (MTED).

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!