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Types and Selection of Cell Culture Media

Source: PricellaPublished: 2023-08-21

Ⅰ. Types of Cell Culture Media

Cell culture medium is not only the basic material for supplying cell nutrition and promoting cell proliferation, but also providing the living environment for cell growth and proliferation.

1. Basal Media


The main components of basal cell culture medium are amino acids, vitamins, carbohydrates, inorganic salts and some other auxiliary substances. When using, a certain amount of serum needs to be added to keep the cells in a good growth state.

There are many types of media, among which DMEM, RPMI 1640, MEM, and DMEM/F12 are widely used.  Others such as M199, IMDM, L15 medium, etc. are also used for the cultivation of some certain cells. The specific features and applications are as follows:


Medium

Features

Applications

BME

BME, designed by Eagle in 1955, added 8 kinds of vitamins and 13 kinds of amino acids on the basis of BSS, with simple ingredients and easy to add.

It is suitable for various cell lines and special research. On this basis, the improved cell culture medium varieties include MEM, DMEM, IMDM, etc.

MEM

MEM is developed by Harry Eagle on the basis of Eagle Medium (BEM). The composition of MEM medium is simple, only containing 12 kinds of essential amino acids, glutamine and 8 kinds of vitamins, mainly used for the culture of adherent cells. When the formula was revised in 1959, lysine and biotin were deleted and the amino acid concentration was increased. The modified recipe can also be used for other types of cell culture.

It is suitable for the growth of a variety of cell monolayers, and is the most basic medium with the widest range of applications. However, due to its limited nutrient content, it is not always the most effective or economical medium to use for specific cell cultures.In addition, MEM cell culture media are available in types containing Earle's balanced salt or Hanks' balanced salt; autoclavable types or filtered sterilized types; and types containing non-essential amino acids. Note that the appropriate MEM cell culture medium should be selected according to actual conditions.

M199

M199 was designed by Morgan in 1950 and contains 53 ingredients except BSS.

With the addition of appropriate amounts of serum, it can be widely used in various cell cultures, as well as in virology, vaccine production, and the culture of rat pancreatic epithelial cells and mouse lens tissue.

DMEM

DMEM is Eagle's medium modified by Dulbecco. DMEM has twice the concentration of amino acids and four times the concentration of vitamins as MEM, with the addition of non-essential amino acids, trace iron ions, and sodium pyruvate.It is divided into low glucose type (1000mg/L) and high glucose type (4500mg/L).

It is commonly used in transformed cell culture for hybridoma bone marrow cells and DNA transfection. For example, CHO cells express and produce hepatitis B vaccine, CHO cells express EPO.

IMDM

IMDM is Lscove's modified Eagle's medium, which is very nutritious with the addition of selenium, HEPES, sodium pyruvate, and additional amino acids and vitamins on the basis of the DMEM medium.

It can be used for rapid culture of hybridoma cells and high-density cells.

RPMI-1640

RPMI-1640 was developed by Moore et al. at Roswell Park Memorial Institute in 1967. It is designed for lymphocyte culture and contains BSS, 21 kinds of amino acids and 11 kinds of vitamins.

It is widely used in the culture of various normal cells and cancer cells, especially the culture of suspension cells.

Ham's F-10, F-12

Ham's F-10 and F-12 were designed by Ham in 1963 and 1969, both containing trace elements. Ham's F-12 is designed on the basis of Ham's F-10 nutrient mixture, which can be used when serum levels are low.

F10 is suitable for hamster and human diploid cells, while F12 is suitable for CHO cells. Ham's F-12 with low serum content is especially suitable for cell culture and cloning culture. It is also widely used in the culture of cancer cells and primary cells after adding serum, such as rat hepatocytes, rat prostate epithelial cells, chondrocytes, rat myoblasts, and chicken embryonic cells.

DMEM/F12

DMEM/F12 is based on DMEM medium with richer nutrients, containing various trace elements.

It is widely used in the culture of various mammalian cells. At the same time, it is often used as a basis for the development of serum-free media, and is also suitable for the culture of mammalian cells at low serum content and clonal density culture.

McCoy's 5A

McCoy's 5A medium was designed by Thomas McCoy et al. in 1959 and originally specialized for the culture of Novikoff Hepatoma cells. McCoy's 5A medium is a modified Basal Medium 5A. Unlike other media, McCoy's 5A contains reduced glutathione, bacto-peptone and high glucose.

It is widely used in many types of primary cell culture, such as bone marrow, skin, gingiva, kidney, spleen, lung, rat embryo, omentum, etc In addition, McCoy's 5A medium is also used for tissue biopsy culture, cell line establishment, and the culture of some lymphocytes and difficult-to-culture cells.

L15

The buffer system of L-15 medium consists of phosphate and free base amino acids, replacing the traditional sodium bicarbonate buffer system. Meanwhile, the replacement of glucose with galactose and sodium pyruvate prevents the formation of acidic metabolic by-products and helps to maintain the stability of culture solution pH, making it suitable for cell culture in non-CO2 equilibrium environments.

It is suitable for the culture of monkey kidney cells and HEP-2, the isolation of primary (such as embryonic and adult tissue) cells, the culture of various viruses, and the culture of neurons, etc.

 

2.  Complete Media


Complete media are the addition of serum, antibiotics and other substances to the basic medium, also called serum media. Most cell cultures are commonly used in bovine serum because it contains various growth factors that promote cell proliferation. In addition, to prevent contamination, a certain amount of antibiotics need to be added to the culture media.

Complete media  can be divided into cell growth media and cell maintenance media for different cells and different research according to the amount of added serum.

3.  Serum-free Media

Serum-free medium refers to the addition of fully defined or partially defined serum replacement components on the basis of synthetic medium, mainly composed of basal medium and additives, namely supplementary factors that replace serum. Its main components are as follows:

Adhesion factors

About 80% cells belong to the adherent growth. General cell culture medium uses serum as its source, and in serum-free medium, cell adhesion factors are very important substances.

Growth Factors

Growth factors are indispensable elements for maintaining cell activity and promoting cell proliferation, migration, and differentiation.

Enzyme Inhibitor

Adherent cells are usually subcultured with trypsin. In general cell culture, serum contains components that inhibit trypsin, so trypsin inhibitors must be added to serum-free medium.

Before the cells transfer to a serum-free medium, the seed cells should be reserved and cultured in the medium with serum as usual to ensure that the characteristics of the cells do not change.

Advantages

1 Few unknown components;
2 It can avoid the toxic effect of serum on cells and serum-derived contamination;
3 Its low content of impurity proteins can avoid serum components from affecting the research results or product manufacturing process, thereby improving the expression level of the product and making it easy to purify.

 

Disadvantages

1 High costs;
2 Not all cells can be grown in serum-free media;
3 Cells are easily affected by some mechanical and chemical factors in serum-free medium, and there are great limitations in medium storage and application.


Ⅱ. Selection of Cell Culture Media

There are many types of cell culture medium, and choosing the right medium is the key to cell culture. Here are some suggestions for reference:

1. If it is a newly purchased cell, you can consult the manufacturer of the cell line, or ask the introduction source to find the corresponding medium on the cell bank;

2. Select the medium according to the characteristics of the cells and the needs of the experiment;

3. Experiment by yourself, culture cells with various media, make a growth curve, record the growth rate of the cells, observe the morphological changes of the cells, and choose the best media according to the experimental results.

There is not a mandatory correspondence between media type and cell line. What medium to use for culturing cells is mainly based on databases, literature and experience.

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