Classic Media and Salts

Basal Salt Mixtures

Dulbecco's Phosphate Buffered Saline
Earle's Balanced Salts
Hanks' Balanced Salts
Tyrode's Salts
Other Salt Mixtures

The use of balanced salt solution (BSS) in cell and tissue culture is generally attributed to early workers in the field. In 1885, Sydney Ringer developed a solution of inorganic salts designed to maintain contractility of mammalian heart tissue. A less specific salt solution was designed by Tyrode for use in work with primary mammalian cells. Tyrode's salt solution became the accepted fluid for diluting protein components of media of natural origin. Since that time, many other balanced salt solutions have been developed for use in cell and tissue culture. The current role of balanced salt solution in cell culture is multi-faceted and can be divided into four principle functions.


 Dulbecco's Phosphate Buffered Saline (D-PBS)

 

Product No. Product Name Description Application
D8662 Dulbecco's Phosphate Buffered Saline
  • Liquid
  • With calcium chloride and magnesium chloride
  • Sterile-filtered
  • Endotoxin tested
  • Cell culture tested
Known as D-PBS, this is a general use, isotonic saline solution for washing cells and tissues.
D1283 Dulbecco's Phosphate Buffered Saline (10x)
  • Liquid
  • With calcium chloride and magnesium chloride
  • pH 4.2-4.6 at 10x
  • Sterile-filtered
  • Endotoxin tested
  • Cell culture tested
Concentrated form of D8662.
D8537 Dulbecco's Phosphate Buffered Saline Modified
  • Liquid
  • Without calcium chloride and magnesium chloride
  • Sterile-filtered
  • Endotoxin tested
  • Cell culture tested
Calcium and magnesium promote cell adhesion. Use this formulation of D-PBS when tissue or cell dissociation or release is the objective.
D1408 Dulbecco's Phosphate Buffered Saline (10x)
  • Liquid
  • Sterile-filtered
  • pH 6.6-7.0 at 10x
  • Sterile-filtered
  • Endotoxin tested
  • Cell culture tested
Concentrated form of D8537.
D5652 Dulbecco's Phosphate Buffered Saline Modified
  • Powder
  • Without calcium chloride and magnesium chloride
  • Formulated at 9.6 grams of powder per liter of medium
  • Related products: Dulbecco's Phosphate Buffered Saline Hybri-Max (D 7030)
  • Cell culture tested
Powder form of D8537.

Calcium and magnesium promote cell adhesion.

Use this formulation of D-PBS when tissue or cell dissociation or release is the objective.
D7030 Dulbecco's Phosphate Buffered Saline Modified Hybri-Max.
  • Powder
  • Without calcium chloride and magnesium chloride
  • Formulated at 9.6 grams of powder per liter of medium
  • Endotoxin tested
  • Hybridoma tested
Same as D5652 except it is tested for use with hybridoma cells.
D5773 Dulbecco's Phosphate Buffered Saline
  • Powder
  • Without calcium chloride
  • Formulated at 9.7 grams of powder per liter of medium
  • Cell culture tested
Calcium promotes cell adhesion.

Use this formulation of D-PBS when tissue or cell dissociation or release is the objective.

Also, suitable for studies that involve calcium metabolism.
D4031 Dulbecco's Phosphate Buffered Saline Modified
  • Liquid
  • With 36 mg/L sodium pyruvate, 50 mg/L streptomycin sulfate, 100 mg/L kanamycin monosulfate, 1000 mg/L glucose, calcium chloride, and magnesium chloride.
  • Sterile-filtered
  • Endotoxin tested
  • Cell culture tested
Fully supplemented isotonic solution.
Contains energy sources and antibiotics.

Frequently used with primary cells where risk of contamination may be high.
D6650 Dulbecco's Phosphate Buffered Saline Modified
  • Powder
  • With 36 mg/L sodium pyruvate, 50 mg/L streptomycin sulfate and 1.000 mg/L glucose.
  • w/o calcium chloride and penicillin G
  • Penicillin G supplied separately with each package for addition at a final concentration of 1 million units/L. Calcium chloride di-hydrate is supplied separately with each package for final concentration of 133 mg/L.
  • Cell culture tested
Isotonic saline solution that contains energy sources and antibiotics.

Provides the option to formulate with or without penicillin G or calcium.

Frequently used with primary cells where risk of contamination may be high and where tissue dissociation is an objective.


 Earle's Balanced Salts (EBSS)

 

Product No. Product Description Application
E2888 Earle's Balanced Salt Solution
  • Liquid
  • With sodium bicarbonate
  • Sterile-filtered
  • Endotoxin tested
  • Cell culture tested
Complete and ready to use formulation of EBSS.
E7510 Earle's Balanced Salt Solution (10x)
  • Liquid
  • Without sodium bicarbonate; supplement with 2.2 gm/L sodium bicarbonate at 1x
  • pH 4.0-4.4 at 10x
  • Sterile-filtered
  • Endotoxin tested
  • Cell culture tested
Concentrated version of E2888.  Must be diluted to 1x, supplemented with sodium bicarbonate and pH adjusted before use.
E6132 Earle's Balanced Salts
  • Powder
  • Without sodium bicarbonate; supplement with 2.2 gm/L sodium bicarbonate
  • Formulated at 8.7 grams of powder per liter of medium
  • Cell culture tested
Powder version of the complete EBSS formulation (E2888).

Sodium bicarbonate is added after the EBSS is dissolved in tissue culture grade water.  The pH is adjusted after addition of sodium bicarbonate.
E3024 Earle's Balanced Salt Solution
  • Liquid
  • With sodium bicarbonate
  • Without phenol red
  • Sterile-filtered
  • Endotoxin tested
  • Cell culture tested
Same as E2888, except without phenol red.

Phenol red has been shown to interfere with the growth of some cells at cloning densities.
Use this medium when working with stem cells or when growing cells at low densities.
E3261 Earle's Balanced Salt Solution Modified
  • Powder
  • Without phenol red and sodium bicarbonate; supplement with 2.2 gm/L sodium bicarbonate
  • Formulated at 8.7 grams of powder per liter of medium
  • Cell culture tested
Powder version of E3024.

Sodium bicarbonate is added after the EBSS is dissolved in tissue culture grade water.  The pH is adjusted after addition of sodium bicarbonate.

Phenol red has been shown to interfere with the growth of some cells at cloning densities.

Use this medium when working with stem cells or when growing cells at low densities.
E6267 Earle's Balanced Salt Solution
  • Liquid
  • With sodium bicarbonate
  • Without calcium chloride, magnesium sulfate
  • Sterile-filtered
  • Endotoxin tested
  • Cell culture tested
Same as E2888, except that calcium and magnesium have been removed.

Calcium and magnesium support cell adhesion.

Recommended for suspension cultures or whenever cell clumping is a problem.
E2899 Earle's Balanced Salt Solution
  • Liquid
  • With sodium bicarbonate
  • Without calcium chloride, magnesium sulfate, and phenol red
  • Sterile-filtered
  • Endotoxin tested
  • Cell culture tested
Same as E6267, except that phenol red has also been removed.

Phenol red has been shown to interfere with the growth of some cells at cloning densities.

Use this medium when working with stem cells or when growing cells at low densities.


 Hanks' Balanced Salts (HBSS)

 

Product No. Product Description Application
H9269 Hanks' Balanced Salt Solution
  • Liquid
  • With sodium bicarbonate
  • Sterile-filtered
  • Endotoxin tested
  • Cell culture tested
Complete and ready to use formulation of HBSS.
H1641 Hanks' Balanced Salt Solution (10x)
  • Liquid
  • Without sodium bicarbonate; supplement with 0.35 gm/L sodium bicarbonate at 1x
  • pH 5.3-6.3 at 10x
  • Sterile-filtered
  • Endotoxin tested
  • Cell culture tested
Concentrated version of H9269. Must be diluted to 1x, supplemented with sodium bicarbonate and pH adjusted before use.
H6136 Hanks' Balanced Salts
  • Powder
  • Without sodium bicarbonate; supplement with 0.35 gm/L sodium bicarbonate 
  • Formulated at 9.8 grams of powder per liter of medium
  • Cell culture tested
Powder version of the complete HBSS formulation (H9269).

Sodium bicarbonate is added after the HBSS is dissolved in tissue culture grade water. The pH is adjusted after addition of sodium bicarbonate.

Same formulation as Hanks' Balanced Salts, Hybri-Max. (H6393).
H6393 Hanks' Balanced Salts, Hybri-Max.
  • Powder
  • Without sodium bicarbonate; supplement with 0.35 gm/L sodium bicarbonate 
  • Formulated at 9.8 grams of powder per liter of medium
  • Endotoxin tested
  • Hybridoma  tested
Related to H6136. Specifically, tested for use with hybridoma cells.
Labeled for in vitro diagnostic use.
H8264 Hanks' Balanced Salt Solution Modified
  • Liquid
  • With sodium bicarbonate
  • Without phenol red
  • Sterile-filtered
  • Endotoxin tested
  • Cell culture tested
Same as H9269, except without phenol red.

Phenol red has been shown to interfere with the growth of some cells at cloning densities.

Use this medium when working with stem cells or when growing cells at low densities.
H1387 Hanks' Balanced Salts Modified
  • Powder
  • Without phenol red and sodium bicarbonate; supplement with 0.35 gm/L sodium bicarbonate 
  • Formulated at 9.8 grams of powder per liter of medium
  • Cell culture tested
Powder version of H8264.

Sodium bicarbonate is added after the HBSS is dissolved in tissue culture grade water. The pH is adjusted after addition of sodium bicarbonate.

Phenol red has been shown to interfere with the growth of some cells at cloning densities.

Use this medium when working with stem cells or when growing cells at low densities.
H9394 Hanks' Balanced Salt Solution Modified
  • Liquid
  • With sodium bicarbonate
  • Without calcium chloride, magnesium sulfate,
  • Sterile-filtered
  • Endotoxin tested
  • Cell culture tested
Same as H6649, except that calcium and magnesium have been removed.

Calcium and magnesium support cell adhesion.

Recommended for suspension cultures or whenever cell clumping is a problem.
H4641 Hanks' Balanced Salt Solution (10x)
  • Liquid
  • Without calcium chloride, magnesium sulfate, and sodium bicarbonate; supplement with 0.35 gm/L sodium bicarbonate
  • pH 5.4-6.4 at 10x
  • Sterile-filtered
  • Endotoxin tested
  • Cell culture tested
Concentrated version of H9394. Must be diluted to 1x, supplemented with sodium bicarbonate and pH adjusted before use.

Calcium and magnesium support cell adhesion.

Recommended for suspension cultures or whenever cell clumping is a problem.
H2387 Hanks' Balanced Salts Modified
  • Powder
  • Without calcium chloride, magnesium sulfate, and sodium bicarbonate; supplement with 0.35 gm/L sodium bicarbonate 
  • Formulated at 9.5 grams of powder per liter of medium
  • Cell culture tested
Powder version of H9394.

Sodium bicarbonate is added after the HBSS is dissolved in tissue culture grade water. The pH is adjusted after addition of sodium bicarbonate.

Calcium and magnesium support cell adhesion.

Recommended for suspension cultures or whenever cell clumping is a problem.
H6648 Hanks' Balanced Salt Solution Modified
  • Liquid
  • With sodium bicarbonate
  • Without calcium chloride, magnesium sulfate, and phenol red
  • Sterile-filtered
  • Endotoxin tested
  • Cell culture tested
Same as H9394, except that phenol red has been removed.

Calcium and magnesium support cell adhesion. Phenol red has been shown to interfere with the growth of some cells at cloning densities.

Use this medium when working with stem cells or when growing cells at low densities.  Use to minimize cell attachment or aggregation.
H4891 Hanks' Balanced Salts Modified
  • Powder
  • Without calcium chloride, magnesium sulfate, phenol red and sodium bicarbonate; supplement with 0.35 gm/L sodium bicarbonate 
  • Formulated at 9.5 grams of powder per liter of medium
  • Cell culture tested
Powder version of H6648.

Sodium bicarbonate is added after the HBSS is dissolved in tissue culture grade water. The pH is adjusted after addition of sodium bicarbonate.

Calcium and magnesium support cell adhesion.

Phenol red has been shown to interfere with the growth of some cells at cloning densities.

Use this medium when working with stem cells or when growing cells at low densities. Use to minimize cell attachment or aggregation.


 Tyrode's Salts

 

Product No. Product Description Application
T2397 Tyrode's Salt Solution
  • Liquid
  • With sodium bicarbonate
  • Sterile-filtered
  • Endotoxin tested
  • Cell culture tested
Complete and ready to use formulation of Tyrode's salt solution.

For cell washing.
T2145 Tyrode's Salts
  • Powder
  • Without sodium bicarbonate; supplement with 1.0 gm/L of sodium bicarbonate
  • Formulated at 9.7 grams of powder per liter of medium
  • Endotoxin tested
  • Sterile-filtered
  • Cell culture tested
Powder version of the complete Tyrode's salts formulation (T2397).

Sodium bicarbonate is added after the salts are dissolved in tissue culture grade water.  The pH is adjusted after addition of sodium bicarbonate.

For cell washing.
T1788 Tyrode's Solution, Acidic
  • Liquid
  • Contains 4.0 gm/L polyvinylpyrrolidone (average molecular  weight 40,000)
  • pH: 2.5 +/- 0.3
  • Shipped in dry ice
  • Sterile-filtered
  • Endotoxin tested
  • Embryo tested
Special formulation of Tyrode's salts (see formulation).

Recommended for removal of the zona pellucida from mouse embryos. 


 Other Salt Mixtures

The following salts solutions are available for use in specific cell culture or related applications. Generally, the use of one of these formulations for an application is specified in publications or technical protocols.
 

Product No. Product Description Application
A3551 Alsever's Solution
  • Liquid
  • Sterile-filtered
  • Endotoxin Tested
  • Cell culture tested
Essentially a solution containing citrate, sodium chloride and glucose.  Provided without bicarbonate, which may be added.

Hypotonic solution for lysing cells.  Especially, red blood cells.
G9779 Gey's Balanced Salt Solution
  • Liquid
  • Sterile-filtered
  • Endotoxin Tested
  • Cell culture tested
A complete balanced salt solution with a unique combination of "salt forms". Contains the same five basic ions as EBSS, HBSS and D-PBS.
K3753 Kreb's-Henseleit Buffer Modified
  • Powder
  • With 2000 mg/L glucose
  • Without calcium chloride and sodium bicarbonate; supplement with 2.1 gm/L of sodium bicarbonate;
  • Formulated at 9.6 grams of powder per liter of medium
  • Cell culture tested
Contains the same components as K4002. However, these ion components are at different concentrations and in different salt forms (see the formulation for details).
K4002 Kreb's-Ringer Bicarbonate Buffer
  • Powder
  • With 1800 mg/L glucose
  • Without calcium chloride and sodium bicarbonate; supplement with 1.26 gm/L sodium bicarbonate
  • Formulated at 9.5 grams of powder per liter of medium
  • Cell culture tested
Bicarbonate buffered isotonic saline containing magnesium, potassium, sodium and phosphate ions.
P2917 Puck's Saline
  • Liquid
  • With sodium bicarbonate
  • Without calcium chloride, magnesium sulfate, potassium phosphate and sodium phosphate
  • Sterile-filtered
  • Endotoxin tested
  • Cell culture tested
A very simple isotonic salt solution. Contains sodium and potassium chloride, glucose and low concentration of bicarbonate buffer (0.35 gm/L).

For in vitro diagnostic use.
  • As an irrigating, transporting and diluting fluid while maintaining intra- and extra-cellular osmotic balance
  • Provides cells with water and certain bulk inorganic ions essential for normal cell metabolism
  • Combined with a carbohydrate, such as glucose, provides the principle energy source for cell metabolism
  • Provides buffering system to maintain the medium within the physiological pH range (7.2-7.6)

Families of basal salt solutions, generally named after the inventor,  have been developed around the early discovery that cells have a basic requirement for five ions: calcium, magnesium, potassium, sodium, and phosphate.  Each balanced salts family of products differs in the concentration and "salt form" of these ions.

Please note that products listed as "Other Salt Solutions" may have uses other than the culturing of cells and that they may not contain all ions listed above.

Probably the three most important basal salt solutions for cell culture are Dulbecco's phosphate buffered saline (D-PBS); Earle's balanced salts (EBSS) and Hanks' balanced salts (HBSS).  Dulbecco's formulation contains no bicarbonate and hence is less susceptible to pH shift during filtration.  Earle's formulation contains 2.2 gm/L of sodium bicarbonate, and Hanks' formulation contains 0.35 gm/L of sodium bicarbonate. Both of these formulations may require pH adjustment after filtration.

D-PBS is offered with and without glucose as an energy source.  The complete formulae for EBSS and HBSS both contain 1.0 mg/L of glucose.