Life Science > Cell Culture > Classical Media & Salts > Basal Salt Mixtures

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 #

Product Name

Description

Application

D 8662

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.

D 1283

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 D 8662.

D 8537

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.

D 1408

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 D 8537.

D 5652

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 D 8537.

Calcium and magnesium promote cell adhesion.

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

D 7030

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 D 5652 except it is tested for use with hybridoma cells.

D 5773

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.

D 4031

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.

D 6650

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 #

Product

Description

Application

E 2888

Earle's Balanced Salt Solution

Liquid
With sodium bicarbonate
Sterile-filtered
Endotoxin tested
Cell culture tested

Complete and ready to use formulation of EBSS.

E 7510

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 E 2888. Must be diluted to 1x, supplemented with sodium bicarbonate and pH adjusted before use.

E 6132

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 (E 2888).

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

E 3024

Earle's Balanced Salt Solution

Liquid
With sodium bicarbonate
Without phenol red
Sterile-filtered
Endotoxin tested
Cell culture tested

Same as E 2888, 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.

E 3261

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 E 3024.

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.

E 6267

Earle's Balanced Salt Solution

Liquid
With sodium bicarbonate
Without calcium chloride, magnesium sulfate
Sterile-filtered
Endotoxin tested
Cell culture tested

Same as E 2888, 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.

E 2899

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 E 6267, 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 #

Product

Description

Application

H 9269

Hanks' Balanced Salt Solution

Liquid
With sodium bicarbonate
Sterile-filtered
Endotoxin tested
Cell culture tested

Complete and ready to use formulation of HBSS.

H 1641

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 H 9269. Must be diluted to 1x, supplemented with sodium bicarbonate and pH adjusted before use.

H 6136

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 (H 9269).

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. (H 6393).

H 6393

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 H 6136. Specifically, tested for use with hybridoma cells.

Labeled for in vitro diagnostic use.

H 8264

Hanks' Balanced Salt Solution Modified

Liquid
With sodium bicarbonate
Without phenol red
Sterile-filtered
Endotoxin tested
Cell culture tested

Same as H 9269, 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.

H 1387

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 H 8264.

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.

H 9394

Hanks' Balanced Salt Solution Modified

Liquid
With sodium bicarbonate
Without calcium chloride, magnesium sulfate,
Sterile-filtered
Endotoxin tested
Cell culture tested

Same as H 6649, 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.

H 4641

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 H 9394. 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.

H 2387

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 H 9394.

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.

H 6648

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 H 9394, 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.

H 4891

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 H 6648.

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 #

Product

Description

Application

T 2397

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.

T 2145

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 (T 2397).

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.

T 1788

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 #

Product

Description

Application

A 3551

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.

G 9779

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.

K 3753

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 K 4002. However, these ion components are at different concentrations and in different salt forms. (see the formulation for details).

K 4002

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.

P 2917

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; and
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.

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