Animal cell culture can be described as in vitro maintenance and propagation of animal cells using a suitable nutrient media. Culturing is a process of growing animal cells artificially. The most important and essential step in animal cell culture is selecting appropriate growth medium for invitro cultivation. The selection of the medium depends on the type of cells to be cultured and also the purpose of the culture. Purpose of animal cell culture can be growth, differentiation, or even production of desired products like pharmaceutical compounds.
Animal cells are cultured using a completely natural media, or an artificial media along with some of the natural products.
In the early years of this in vitro cultivation of animal cell culture technique natural media are obtained from biological sources were used. For eample
1. Body fluid such as plasma, serum, lymph, amniotic fluid and much more are used. These fluids used as animal cell culture media after testing for toxicity and sterility.
2. Tissue extract such as extract of liver, spleen, bone marrow and leucocyes also used as animal cell culture media. But most commonly used tissue extract is from chick embryo.
3. Plasma clots are also used as media for animal cell culture and now they are commercially produced as culture media.
4. Bovine embryo extract are also prepared using bovine embryos of up to 10days age, and are used as animal cell culture media.
1. The artificial media contains partly or fully defined components.
2. The basic criteria for choosing a artificial media for animal cell culture are
The culture media should provide all the required nutrients to the cell.
3. Media should maintain the physiological pH at around 7 with the help of buffering system.
4. The animal cell culture media should be sterile, and isotonic to the culturing cells. The basis for the animal cell culture media is the balanced salt solution, which are used to create a physiological pH and osmolarity required to maintain the animal cells in vitro or in laboratory conditions.
5. For promoting cell growth and proliferation, many types of animal cell culture media are designed by adding or varying different constituents. For example serum containing media and serum-free media.
1. Serum media is an example for natural media. Natural media are very useful and convinient for a wide range of animal cell culture. But they also have got some disadvantages such as poor reproductability due to lack of knowledge of exact composition of these natural media.
2. Major reasons for using synthetic media are for immediate survival of cells, for prolonged survival, for indefinite growth and also for specialized functions.
a. Balanced diet solution with specific osmotic pressure and pH are used for the immediate survival
b. Serum or balanced salt solution along with amino acids, oxygen, vitamins and serum proteins are used for long survival.
c. Minimum Essential Medium also known as Eagles media are used for mammalian cell culture
d. Role of serum in animal cell culture is very complex and also it contains mixture of many biomolecules such as growth factors and also growth inhibitory factors.
Function of serum:
1. Serum is bound to proteins and basic nutrient in solution.
2. Serum also contains hormones and growth factors, which play a major role in stimulating cell growth and function
3. Serum also helps in attachment of the cells
4. Serum also acts as spreading factor.
5. Serum also function as binding protein such as albumin carrying factor
6. Serum also helps in carrying hormones, vitamins, minerals, lipids and much more biological substances.
7. Serum also minimizes mechanical damage and also damage caused by viscosity
8. Serum also acts as natural buffering agent and helps in maintaining the pH of the culture media.
Disadvantage of Serum in Media:
1. Serum may contain inadequate amount of cell specific growth factors and may need to suppliment the media or it may also contain in abundance of cytotoxic compounds.
2. Serum media has got high risk of contamination with virus, fumgi and mycoplasma.
3. There is no uniformity in the composition of serum, as still we do not know the exact composition of the serum
4. Special tests are done to maintain the quality of the each batch of serum before it is used in cell culture media.
5. Serum may also contain some of the growth inhibiting factors, these in turn will inhibit the cultured cell growth and proliferation.
6. Serum availability is restrited as they are extracted from cattle
7. Presence of serum in the culture media may interfere with the purification and isolation of cell culture products, such as pharmaceutical compounds. Due to this additional steps are added for the isolation of cell culture products.
Serum Free Media:
As using serum in animal cell culture media has got some disadvantages, to overcome this serum free media are designed and developed.
Advantages of Serum Free Media:
1. Main and important advantage of serum free media is scientists or researchers can control the growth of cultured cells as required by changing the composition of the media.
2. Serum free media can be designed using specific factors, which will help in differentiation of cultured cells with specific desired functions.
Disadvantages of Serum Free Media:
1. cell proliferation is very slow in serum free media.
2. Cultured cells may need more than one type of media to obtain desired cell culture products.
3. Purity of reagents used in the serum free media
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