Overview: Cell Culture Media

Cell culture is an important technique within life science research, in which cells isolated from host organisms are upkept, studied, and manipulated in an artificial environment. These cells are often grown in a gel or liquid-based growth medium comprised of a varying components that support cell proliferation and survival. Cell culture media provide the necessary energy sources, growth factors, hormones, amino acids, vitamins, inorganic salts, and trace elements to maintain the pH and osmotic pressure to keep the target cell lines alive. By artificially maintaining the homeostasis of isolated cells, they can be utilized for a variety of biological experiments, including viability and proliferation assays, cell-based drug screening assays, or cell signaling and gene expression assays. In supporting a variety of scientific techniques, cell culture media hold a large impact in cell biology and development of drugs or gene therapies.

What makes good cell culture media?

Cell culture media are the primary source of nutrients to isolated cells grown in an environment. Without media, cell culture cannot be performed. For optimal results, the media used should contain essential ingredients in appropriate quantities. These essential ingredients often include but are not limited to:

  1. A carbon and energy source: Carbohydrates are the major source of energy in the cell culture media. Even a molecule as simple as glucose can provide the energy to fuel the cellular metabolism of many cell types. Most media contain glucose as a major energy source, but it may be replaced with other sources such as galactose, maltose or sucrose depending upon the cell type and the experimental conditions or requirements.
  2. Buffering system: Regulating and maintaining pH is important for the efficient growth of cells in culture media, which is generally accomplished by growing the cells in a gaseous environment consisting of 5% carbon dioxide. Zwitterionic buffers such as HEPES with highly effective buffering capacity can also be used in cell culture media to maintain the ideal pH for cell growth., which is independent of the cell growth in a gaseous environment. This will prevent the uncontrolled increase or decrease in hydrogen ion concentration throughout experimental use, prolonging the life of viable cells.
  3. pH indicator: Many cell culture medium types contain pH indicators such as Phenol Red to indicate changes in pH throughout their experiments, helping in assessing the culture growth conditions and to determine if resulting cells are viable.
  4. Serum: Commonly fetal bovine serum extracted from bovines, serum is an essential component of many medium types, as it provides a source for proteins, amino acids, carbohydrates, lipids, growth factors, hormones, and several other ingredients including minerals and trace elements.
  5. Amino acids, inorganic salts, minerals: A common similarity in cell culture media is the presence of amino acids, inorganic salts, and minerals, these provide the cells in question with the necessary tools for homeostasis. These ingredients are provided by the serum in the media or can be supplemented depending upon the requirements. Inorganic salts can often be added via balanced salt solutions.
  6. Antibiotics: Antibiotics in culture media provide increased security in ensuring cell culture media are not contaminated with bacterial or fungal strains. By incorporating antibiotics in the media, you can target invasive microbes while preserving the cultured cells.

The type of Cell Culture Media needed varies based on its intended use. Although all types require nutrients to support target cell lines, there are three basic classes of cell culture media that are most prevalent in the life science industry. These classes of media often fall under the following categories[1]

  • Basal media: Basal media often contain amino acids, vitamins, inorganic salts, and a carbon source. This media type can support a wide range of cells and is often supported by further growth factors and serum. Some examples of Basal Media include Terrific Broth, or Lennox Broth.
  • Reduced-serum media: Reduced-serum medium types often contain similar formulations to basal media, with a cap on the levels of serum used. Serum levels often fall in the 2%-10% range or less. Instead of serum, these formulations are enriched with other animal derived factors such as albumin, transferrin, or insulin. Reduced-serum media can often be custom-made depending on the application of interest, and relative serum amount desired.
  • Serum-free media: In serum-free media, the serum typically found in Basal Media and Reduced-Serum Media is replaced with hormonal formulations such as growth factors or cytokines. Although formulations are more complex, serum-free media types offer a defined and consistent environment for cell growth. Serum-free media can often be custom-made depending on the application of interest and can be constructed using pre-calculated components.

These cell culture medium classes differ in their serum content, a significant part of their formulations. Serum is the fluid that is obtained after the clotting of blood. It can also be obtained after clotting factors are removed from a host organism’s blood plasma and is a common supplement in cell culture media as it provides proteins, hormones, lipids, minerals, and growth factors. Although serum is beneficial for a vast majority of application types, the user of the media in unable to define a numerical concentration of components within the serum used, making specific reproducibility more difficult for basal and Reduced-serum media, despite effective results. Each category of cell culture medium types can also sustain a variety of organisms, including bacteria, yeast and fungi, and mammalian cell types. Such cell types have varying growth requirements in their media formulations.

Based on target cell types, the following cell culture mediums can be used:

 

Bacterial Cell Culture Media
Luria-Bertani Broth
(LB Broth) 		General-purpose for E. coli. Used for routine cultivation and protein expression.
M9 Minimal Medium Minimal medium for E. coli. Used for selective growth and metabolic studies and requires specific nutrient supplementation.
Terrific Broth
(TB Broth) 		Rich medium for high-density bacterial cultures. Used for protein expression.
Super Optimal Broth
(SOB Broth) 		Rich medium used to provide an optimal environment for cellular growth and metabolism. Contains higher levels of amino acids than typical basal media.
Super Optimal Broth with Catabolite repression
(SOC Medium) 		Recovery medium after heat shock transformation. Used to enhance plasmid recovery and cell growth.
Yeast/Fungal Cell Culture Media
Yeast Extract-Peptone-Dextrose
(YPD Broth) 		Rich medium for S. cerevisiae and other yeast. Supports growth and maintenance.
Synthetic Defined Medium
(SD Medium) 		Defined medium for yeast that is customizable for specific nutrient requirements. Suitable for selective growth.
Yeast Nitrogen Base
(YNB Medium) 		Minimal medium for yeast that requires specific nutrient supplementation.
Sabouraud Dextrose Broth Fungal culture medium for isolation and cultivation of dermatophytes and yeasts, containing dextrose and peptone.
Mammalian Cell Culture Media
Dulbecco's Modified Eagle's Medium
(DMEM Medium) 		Widely used for suspension of mammalian cells. DMEM contains essential nutrients, vitamins, and glucose and is suitable for various cell lines.
Ham’s F-12 A nutrient mixture made with glucose, amino acids, and inorganic salts, with higher levels of amino acids and sodium pyruvate. Used to cultivate a variety of mammalian and hybridoma cells when combined with serum.
RPMI 1640 Medium Developed for human cells and is suitable for a variety of mammalian cells. Contains amino acids, vitamins, and inorganic salts.
F-12 Media A mixture of DMEM and Ham’s F-12, containing glucose, amino acids, vitamins, and inorganic salt.
IMDM Media Modified version of DMEM with additional selenium, amino acids, vitamins, and glutamine. It also lacks iron.
Medium-199 Composed of purines, pyrimidines, and fat-soluble compounds. Typically used for cultivation of non-transformed cells.
Fetal Bovine Serum
(FBS) 		Used in mammalian cell culture, providing growth factors, hormones, and nutrients. Commonly added to basal media.
Minimum Essential Medium
(MEM) 		General-purpose medium for various mammalian cell lines that contains essential nutrients and vitamins.
Serum-Free Medium
(SFM) 		Designed for cell culture without serum. SFM eliminates serum-derived variability and is suitable for specific applications.

Cell Culture Media at Boston BioProducts

Every cell culture medium is unique to the cell type used and the experimental application. Select the appropriate Cell Culture Medium from the catalog or design your optimal formulation with custom manufacturing options at Boston BioProducts.

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