The 9 best biological buffers for cell culture
One of the most important characteristics of Good’s Buffers is that they are not toxic towards cells. Therefore, these chemicals are widely used in cell culture to maintain the pH of experiments under control.
Due to their special features, many Good’s buffers / biological buffers are considered ideal for the isolation of cells, cell cultivation, enzyme assays, and numerous other biochemical applications.
We collected academic references in which biological buffers were used for cell culture in order to help you understanding which buffers are more suitable for your applications. Here is our list of 9 options you might consider for your research:
Further reading:What are the precautions in Cell Culture research?
1) HEPES
Useful pH range: 6.8 - 8.2pKa (25°C): 7.45 - 7.65
Ion binding: Negligible metal ion binding
How has it been used in cell culture?
- Used to buffer mammalian cell cultures in open systems of high density cultures and in closed systems of high and low density cultures1
- Used as a buffer for in vitro fertilization and embryo culture2,3
Further reading:Why use HEPES?
2) MOPS
Useful pH range: 6.5 - 7.9pKa (25°C): 7.0 - 7.4
Ion binding: Strong interaction only with Fe
How has it been used in cell culture?
- Many studies report its use in cell culture media for bacteria (enterobacteria4, Lactococcus lactis5 and Legionella pneumophila6, for example)
- Used in cell culture media for yeast and mammalian cells. Notice that only concentrations lower than 20mM are suitable for mammalian cell work7
- Used in cell culture media to study the meiotic regulation in mouse oocytes8
Further reading:8 uses of MOPS buffer you didn't know
3) MES
pKa (25°C): 5.9 - 6.3
Ion binding: Strong interaction only with Fe
How has it been used in cell culture?
- Some studies report its use in cell culture media for the growth of bacteria (halophilic bacteria, for example)9
- Used in cell culture media for yeast and mammalian cells. Notice that only concentrations lower than 20mM are suitable for mammalian cell work7
- Used in plant culture media (but at concentrations around 10mM – it is toxic to most plants at higher concentrations)10
- Used in culture media to initiate the growth of pine and fir trees11
Further reading:15 uses of MES Buffer you didn't know
4) BES
pKa (25°C): 6.9 - 7.3
Ion binding: Strong interaction only with Cu
How has it been used in cell culture?
- Used in culture media for bacteriophage adsorption12
5) MOPSO
Useful pH range: 6.2 - 7.6pKa (25°C): 6.7 - 7.1
Ion binding: Strong interaction only with Fe
How has it been used in cell culture?
- Used as a buffer component of charcoal yeast extract medium for the growth of the bacteria Legionella pneumophila6
6) ACES
Useful pH range: 6.1 - 7.5pKa (25°C): 6.6 - 7.0
Ion binding: Strong interaction with Cu and Mg
How has it been used in cell culture?
- Used as a buffer component of charcoal yeast extract medium for the growth of the bacteria Legionella pneumophila6
- Used in cell culture media for hairy roots of Catharanthus roseus, a plant commonly known as rose periwinkle13
7) TAPS
Useful pH range: 7.7 - 9.1pKa (25°C): 8.25 - 8.65
Ion binding: Strong interaction with Cu, Cr and Fe
How has it been used in cell culture?
- Used in cell culture media for experiments with dinoflagellates (e.g.: marine plankton)14
8) Bicine
pKa (25°C): 8.1 - 8.5
Ion binding: Strong interaction with Cu, Fe, Co, Mg, Ca, Ni, Zn and Cd
How has it been used in cell culture?
- Used for the culture of ammonia fungi15
9) Tricine
Useful pH range: 7.4 - 8.8pKa (25°C): 8.0 - 8.3
Ion binding: Strong interaction with Mg, Ca, Co, Cu, Ni and Zn
How has it been used in cell culture?
- Used in culture media of bacteria to prevent precipitation of iron salts4
- Used in animal tissue culture16
Further reading:Why use TRICINE Buffer
References:
1 Blanchard, J.S. (1984) Methods Enzymol. 104, 404-414. Available at https://www.ncbi.nlm.nih.gov/pubmed/6717292
2 Alonso, A. D. C., Zaidi, T., Novak, M., Grundke-Iqbal, I., and Iqbal, K. (2001) Hyperphosphorylation induces self-assembly of τ into tangles of paired helical filaments/straight filaments. Proceedings of the National Academy of Sciences, 98(12), 6923-6928. Available at https://www.ncbi.nlm.nih.gov/pmc/articles/PMC34454/
3 Kashino, Y., Koike, H., and Satoh, K. (2001) An improved sodium dodecyl sulfate-polyacrylamide gel electrophoresis system for the analysis of membrane protein complexes. Electrophoresis, 22(6), 1004-1007. Available at https://onlinelibrary.wiley.com/doi/abs/10.1002/1522-2683%28%2922%3A6%3C1004%3A%3AAID-ELPS1004%3E3.0.CO%3B2-Y
4 Neill, S. J., Desikan, R., Clarke, A., and Hancock, J. T. (2002) Nitric oxide is a novel component of abscisic acid signaling in stomatal guard cells. Plant Physiology, 128(1), 13-16. Available at https://www.plantphysiol.org/content/128/1/13
5 Grady, J.K. et al. (1988) Anal. Biochem. 173, 111-115. Available at https://www.ncbi.nlm.nih.gov/pubmed/2847586
6 Parfitt, D. E., Almehdi, A. A. and Bloksberg, L. N. (1988) Sci. Hortic., 36, 157–163. Available at https://www.sciencedirect.com/science/article/pii/0304423888900490
7 Ferreira, C.M., Pinto, I.S., Soares, E.V., Soares, H.M. (2015) (Un)suitability of the use of pH buffers in biological, biochemical and environmental studies and their interaction with metal ions – a review, Royal Society of Chemistry, 30989- 31003. Available at https://repositorium.sdum.uminho.pt/bitstream/1822/38712/1/document_19948_1.pdf
8 Nagira, K., Hayashida, M., Shiga, M., Sasamoto, K., Kina, K., Osada, K., Sugahara, T. and Murakami, H. (1995) Cytotechnology, 17, 117–125. Available at https://link.springer.com/article/10.1007/BF00749399
9 Soares, E. V., Duarte, A. P. R. S. and Soares, H. M. V. M. (2000) Chem. Speciation Bioavailability, 12, 59-65
10 Xu, X; Khan, M. K.; Burgess, D. J. (2012) A Two-Stage Reverse Dialysis In Vitro Dissolution Testing Method for Passive Targeted Liposomes, Int. J. Pharm., 426, 211–218. Available at https://www.sciencedirect.com/science/article/pii/S0378517312000646?via%3Dihub
11 Taha, M., Gupta, B. S., Khoiroh, I., Lee, M-J. (2011) Interactions of Biological Buffers: The Ubiquitous “Smart” Polymer PNIPAM and the Biological Buffers, MES, MOPS and MOPSO. Macromolecules. 44, 8575-8589. Available at https://pubs.acs.org/doi/abs/10.1021/ma201790c
12 Zhao, G., and Chasteen, N. D. (2006) Anal. Biochem., 349, 262–267. Available at https://www.ncbi.nlm.nih.gov/pubmed/16289439
13 Koerner, M. M., Palacio, L. A., Wright, J. W., Schweitzer, K. S., Ray, B. D. and Petrache, H. I (2011) Biophys. J., 101, 362–369. Available at https://www.ncbi.nlm.nih.gov/pubmed/21767488
14 Bevans, C. G. and Harris, A. L. (1999) J. Biol. Chem., 274, 3711–3719. Available at https://www.jbc.org/content/274/6/3711
15 Baker, C. J., Mock, N. M., Roberts, D. P., Deahl, K. L., Hapeman, C. J., Schmidt, W. F. and Kochansky, J. (2007) Free Radical Biol. Med., 43, 1322–1327. Available at https://europepmc.org/abstract/med/17893045