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B-27 Neuronal Culture Supplements

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Choosing the right B-27 supplement for neuronal culture

 

More than 30 years ago, B-27 Neuronal Culture Supplement was designed and optimized by Dr. Greg Brewer to improve survival of primary neurons in culture and became the first commercially available serum-free neuronal cell culture supplement. Since then, the B-27 portfolio has expanded to support a broad range of neuronal and advanced cell culture applications.

 

Choose the right formulation for neuronal cell culture

  • B-27 Supplement is a trusted, widely adopted serum-free supplement supporting primary neurons, neural stem cells, PSC-derived neurons, 3D cell culture, disease modeling, and more.
  • B-27 Plus Supplement is an optimized, next-generation formulation that builds on the trusted performance of B-27 Supplement to help researchers advance neuronal cell culture models.
  • B-27 Plus Neuronal Culture System combines B-27 Plus Supplement with Neurobasal PhysioPlus Medium to support improved neuronal survival, maturation, and long-term culture, helping researchers develop more robust and physiologically relevant neuronal models.
  • Trusted performance, continued innovation, from the original B-27 Supplement to B-27 Plus Supplement, the portfolio combines decades of trusted performance with ongoing innovation to support evolving cell culture needs.

Why is B-27 among the most trusted supplements for healthy cell culture of many different cell types?

Helping researchers be productive, publish, and make scientific breakthroughs

  • Designed for dependable consistency—every lot is performance-tested for reliable results
  • High quality—cGMP media manufacturing allows for the highest standards of quality
  • Versatile usage—ideal for use with serum-free media such as Neurobasal Medium or Neurobasal-A Medium, without the need for an astrocyte feeder layer. Supports the growth and viability of neurons, as well as many other cell types differentiated from PSCs.
  • Trusted performance—a proprietary formulation; the first and most cited neuronal cell culture supplement, trusted by researchers worldwide

B-27 Supplement vs. other suppliers’ supplements

Figure 1. B-27 Supplement system supported more live cells per mm2 in primary mouse and rat cortical neurons compared to competitor products. Traditional culture of primary rodent neurons in Neurobasal Medium (unless noted otherwise). The B-27 Supplement better supports healthy neurons than supplements from other suppliers.

Download our free comprehensive infographic to discover how B-27 Supplement can enhance your research.

Learn how advanced cryopreservation techniques and Gibco solutions can help you confidently preserve and recover functional neurons for your next breakthrough in neuroscience.

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 produces 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.

Trusted history, innovative future: discover the full range of B-27 supplement solutions

For more than three decades, B-27 Supplement has been the most widely used serum-free growth supplement, and we continue to innovate and optimize it to enable future advancements in cell culture and allow customers to create advanced cell models. We offer a variety of B-27 Supplement formulations, with B-27 Plus Supplement designed to support enhanced cell survival, maturation, and maintenance of neurons.

Recommendations for neuronal applications for B-27 supplement family

Application description

B-27 version(s) used

Cat. No.

Cell therapy manufacturing applications

CTS B-27 Supplement, Xeno-free

A5047501, A5047601, A5047701

Differentiation of human PSCs/NSCs to neurons

B-27 Supplement

17504044

Expansion of neural stem cells (NSCs) or neural progenitor cells (NPCs)

B-27 Supplement without vitamin A

12587010

Functional maturation and electrophysiology studies such as neural network activity assays, MEA, or patch-clamp recordings

B-27 Plus Supplement or B-27 Plus Neuronal Culture System

A3582801 or A40008171

Maturation and maintenance of human PSC-derived neurons

B-27 Plus Supplement or B-27 Plus Neuronal Culture System

A3582801 or A40008171

Maturation and maintenance of primary neurons (rodent and human)

B-27 Plus Supplement or B-27 Plus Neuronal Culture System

A3582801 or A40008171

Studies of insulin secretion, insulin receptors, or insulin/IGF signaling

B-27 Supplement without insulin

A1895601

Studies of oxidative stress/damage, apoptosis, or where free radical damage to neurons occurs

B-27 Supplement without AO (antioxidants)

10889038

Interested in regenerative medicine and cell therapy applications? 

Take your neuronal cell culture further with B-27 Plus Neuronal Culture System

 

The next-generation Gibco B-27 Plus Neuronal Culture System, composed of B-27 Plus Supplement and Neurobasal PhysioPlus Medium, supports earlier, sustained neuronal activity, and excellent cell viability to help accelerate scientific discoveries. Together, these small changes yield big results in the maintenance and maturation of primary neurons to offer the following improved benefits:

  • Enable a higher proportion of electrically active neurons to be maintained across the cell population of the culture
  • Promote early and prolonged maintenance of spontaneous neuronal activity that doesn’t drop off during multi-week experiments
  • Support sustained and stable network activity during long-term culture, increasing the useful experimental window of neuronal cultures

Advancing 3D neural cell culture with B-27 supplement

Complex 3D cell culture models are increasingly used to study development and disease, offering the potential to more closely recapitulate in vivo tissue architecture and physiology than traditional 2D cultures. B-27 Supplement has become widely used across 3D neural culture workflows, including both unguided cerebral organoids and guided neural organoid models.

 

With its trusted ability to support neuronal growth and differentiation, B-27 Supplement is incorporated into methods and protocols spanning neural induction, expansion, organoid formation, and maturation. Its utility also extends beyond neural models, with demonstrated use in 3D culture workflows for cardiac, liver, intestinal, pancreatic, and other tissue models.

B-27 Supplement supports extensive range of applications

Application

B-27 version(s) used

Cat. No.

Derivation of cardiac cells/cardiomyocytes from iPSCs1,2,3

B-27 Supplement without insulin (early stages) and B-27 Supplement (later stages)

17504044 or A1895601

Derivation of gut organoids and PSC-derived intestinal/colon cells4,5,6

B-27 Supplement or B-27 Supplement without Vitamin A

17504044 or 12587010

Derivation of lung organoids from PSCs or adult tissue7,8,9

B-27 Supplement or B-27 Supplement without Vitamin A

17504044 or 12587010

Derivation of unguided cerebral organoids and guided region-specific neural organoids10,11

B-27 Supplement or B-27 Supplement without Vitamin A

17504044 or 12587010

Differentiation of PSCs into pancreatic cells or beta islets and expansion of adult pancreas progenitors12,13,14

B-27 Supplement or B-27 Supplement without Vitamin A

17504044 or 12587010

Differentiation of PSCs to kidney cells and nephron-like kidney organoids15,16

B-27 Supplement

17504044

Establishment of adult liver organoids and PSC-derived hepatic cells17

B-27 Supplement without Vitamin A or B-27 Supplement

17504044 or 12587010

Maintenance of naïve human PSC cultures in N2B27-based, 2iL+Gö, 5iLAF, t2iLGöY or PXGL media18,19

B-27 Supplement

17504044

Retinal organoids/retinal differentiation, maintenance of retinal cells, including retina pigment epithelium cells (RPE) and retinal ganglion cells (RGCs)20, 21, 22, 23

B-27 Supplement, B-27 Supplement without Vitamin A, or B-27 Supplement for RGCs

17504044 or 12587010 or 0080085SA

Serum-free mouse embryonic stem cell (mESC) self-renewal in N2B27/2i/LIF ground-state culture24

B-27 Supplement or B-27 Supplement minus vitamin A

17504044 or 12587010
  1. Kempf H, Kropp C, Olmer R, Martin U, Zweigerdt R. Cardiac differentiation of human pluripotent stem cells in scalable suspension culture. Nat Protoc. 2015 Sep;10(9):1345–1361. doi: 10.1038/nprot.2015.089. Epub 2015 Aug 13.
  2. Batalov I, and Feinberg AW. Differentiation of Cardiomyocytes from Human Pluripotent Stem Cells Using Monolayer Culture. Biomark Insights. 2015 May 27;10(Suppl 1):71–76. doi: 10.4137/BMI.S20050.
  3. Lian Z, Zhang J, Azarin SM, et al. Directed cardiomyocyte differentiation from human pluripotent stem cells by modulating Wnt/β-catenin signaling under fully defined conditions. Nat Protoc. 2013 Jan;8(1):162–175. doi: 10.1038/nprot.2012.150. Epub 2012 Dec 20.
  4. Spence JR, Mayhew CN, Rankin SA, et al. Directed differentiation of human pluripotent stem cells into intestinal tissue in vitro. Nature. 2011;470(7332):105–109. doi: 10.1038/nature09691.
  5. McCracken KW, Catá EM, Crawford CM, et al. Modelling human development and disease in pluripotent stem-cell-derived gastric organoids. Nature. 2014;516(7531):400–404. doi: 10.1038/nature13863.
  6. Sato T, Stange DE, Ferrante M, et al. Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett's epithelium. Gastroenterology. 2011;141(5):1762–1772. doi: 10.1053/j.gastro.2011.07.050.
  7. Thermo Fisher Scientific Protocol: A simplified protocol for the development of 3D lung organoids.
  8. Dye B, Hill D, Ferguson MAH, et al. In vitro generation of human pluripotent stem cell derived lung organoids. Elife. 2015 Mar 24;4:e05098. doi: 10.7554/eLife.05098.
  9. Salahudeen AA, Choi SS, Rustagi A, et al. Progenitor identification and SARS-CoV-2 infection in human distal lung organoids. Nature. 2020 Dec;588(7839):670–675. doi: 10.1038/s41586-020-3014-1. Epub 2020 Nov 25.
  10. Giandomenico SL, Sutcliffe M, Lancaster MA. Generation and long-term culture of advanced cerebral organoids for studying later stages of neural development. Nat Protoc. 2021 Feb;16(2):579–602. doi: 10.1038/s41596-020-00433-w. Epub 2020 Dec 16.
  11. Miura Y, Li M-Y, Revah O, et al. Engineering brain assembloids to interrogate human neural circuits. Nat Protoc. 2022 Jan;17(1):15–35. doi: 10.1038/s41596-021-00632-z. Epub 2022 Jan 6.
  12. D'Amour KA, Bang AG, Eliazer S, et al. Production of pancreatic hormone-expressing endocrine cells from human embryonic stem cells. Nat Biotechnol. 2006;24(11):1392–1401. doi: 10.1038/nbt1259.
  13. Huch M, Bonfanti P, Boj SF, et al. Unlimited in vitro expansion of adult bi-potent pancreas progenitors through the Lgr5/R-spondin axis. EMBO J. 2013;32(20):2708–2721. doi: 10.1038/emboj.2013.204.
  14. Li X, Ma Z, Lui KO. A differentiation protocol for the generation of pancreatic beta-like cells from human embryonic stem cells. STAR Protoc. 2023 Sep 15;4(3):102407. doi: 10.1016/j.xpro.2023.102407. Epub 2023 Jun 30.
  15. Musah S, Dimitrakakis N, Camacho DM, et al. Directed differentiation of human induced pluripotent stem cells into mature kidney podocytes and establishment of a Glomerulus Chip. Nat Protoc. 2018 Jul;13(7):1662–1685. doi: 10.1038/s41596-018-0007-8.
  16. Freedman BS, Brooks CR, Lam AQ, et al. Modelling kidney disease with CRISPR-mutant kidney organoids derived from human pluripotent epiblast spheroids. Nat Commun. 2015 Oct 23;6:8715. doi: 10.1038/ncomms9715.
  17. Broutier L, Andersson-Rolf A, Hindley CJ, et al. Culture and establishment of self-renewing human and mouse adult liver and pancreas 3D organoids and their genetic manipulation. Nat Protoc. 2016 Sep;11(9):1724–1743. doi: 10.1038/nprot.2016.097. Epub 2016 Aug 25.
  18. Moya-Jodar M, Ullate-Agote A, Barlabe P, et al. Revealing cell populations catching the early stages of human embryo development in naive pluripotent stem cell cultures. Stem Cell Reports. 2023 Jan 10;18(1):64–80. doi: 10.1016/j.stemcr.2022.11.015. Epub 2022 Dec 22.
  19. Bredenkamp N, Yang J, Clarke J, et al. Wnt inhibition facilitates RNA-mediated reprogramming of human somatic cells to naïve pluripotency. Stem Cell Reports. 2019;13(6):1083–1098. doi: 10.1016/j.stemcr.2019.10.012.
  20. Salero E, Blenkinsop TA, Corneo B, et al. Adult Human RPE Can Be Activated into a Multipotent Stem Cell that Produces Mesenchymal Derivatives. Cell Stem Cell. 2012 Jan 6;10(1):88–95. doi: 10.1016/j.stem.2011.11.018.
  21. Gill KP, Hung SSC, Sharov A, et al. Enriched retinal ganglion cells derived from human embryonic stem cells. Sci Rep. 2016 Aug 10;6:30552. doi: 10.1038/srep30552.
  22. Rowland TJ, Blaschke AJ, Buchholz DE. Differentiation of human pluripotent stem cells to retinal pigmented epithelium in defined conditions using purified extracellular matrix proteins. J Tissue Eng Regen Med. 2013 Aug;7(8):642–653. doi: 10.1002/term.1458. Epub 2012 Apr 18.
  23. Zhong X, Gutierrez C, Xue T, et al. Generation of three-dimensional retinal tissue with functional photoreceptors from human iPSCs. Nat Commun. 2014 Jun 10;5:4047. doi: 10.1038/ncomms5047.
  24. Ying Q-L, Wray J, Nichols J, et al. The ground state of embryonic stem cell self-renewal. Nature. 2008 May 22;453(7194):519–523. doi: 10.1038/nature06968.

 

B-27™ is a registered trademark of the Board of Trustees of Southern Illinois University used under exclusive license by Thermo Fisher Scientific.

For Research Use Only. Not for use in diagnostic procedures.