The 9 best biological buffers for cell culture

forward from: https://www.hopaxfc.com/en

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:

1) HEPES

Useful pH range: 6.8–8.2
pKa (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 cultures(1)
• Used as a buffer for in vitro fertilization and embryo culture(2,3)

Read more about Hopax HEPES

2) MOPS

Useful pH range: 6.5–7.9
pKa (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 (enterobacteria(4), Lactococcus lactis(5) and Legionella pneumophila(6), for example)
• Used in cell culture media for yeast and mammalian cells. Notice that only concentrations lower than 20mM are suitable for mammalian cell work(7)
• Used in cell culture media to study the meiotic regulation in mouse oocytes(8)

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3) MES

Useful pH range: 5.5–6.7
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 work(7)
• 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 trees(11)

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4) BES

Useful pH range: 6.4–7.8
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 adsorption(12)

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5) MOPSO

Useful pH range: 6.2–7.6
pKa (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

Read more about Hopax MOPSO

6) ACES

Useful pH range: 6.1–7.5
pKa (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 periwinkle(13)

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7) TAPS

Useful pH range: 7.7–9.1
pKa (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)

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8) Bicine

Useful pH range: 7.6–9.0
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 fungi(15)

Read more about Hopax Bicine

9) Tricine

Useful pH range: 7.4–8.8
pKa (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 culture(16)

Read more about Hopax Tricine

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 http://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 http://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 http://europepmc.org/abstract/med/17893045