ANIMAL CELL CULTURE
33. ANIMAL CELL CULTURE
Animal cell culture can be described as invitro maintenance and propagation of animal cells using suitable nutrient media. Purpose of animal cell culture can be growth, differentiation or even production of desired products like pharmaceutical compounds.
33.1. Difference between normal and transformed cells:-
33.2. Primary and Secondary cultures:-
33.3. Cell lines
The various types of cell lines are categorized into two types as Finite cell lines and Continuous cell lines.
Finite cell lines are those cell lines which have a limited life span and grow through a limited number of cell generations. The cells normally divide 20 to 100 times before extinction.
Cell lines transformed under in-vitro conditions give rise to continuous cell lines. The continuous cell lines are transformed, immortal and tumorigenic.
33.4. Requirements for Animal Cell Culture
Among the essential requirements for animal cell, culture is special incubators to maintain the level of oxygen, CO2, temperature, humidity as present in the animal's body. The synthetic media with vitamins, amino acids and fetal calf serum. Following parameters are essential for successful intimate cell culture:-
33.4.1. Temperature:- In most of the mammalian cell cultures, the temperature is maintained at 37°C in the incubators as the body temperature of Homosapiens is 37°C.
33.4.2. Culture media:- The culture media is prepared in such a way that it provides.
1. The optimum conditions of factors like pH, osmotic pressure etc.
2. It should contain chemical constituents which the cells or tissues are incapable of synthesizing generally the media is the mixture of inorganic salts and other nutrients capable of sustaining cells in cultures such as amino acids, fatty acids, sugars, ions, trace elements, vitamins, cofactor and ions. Glucose is added as an energy source. Phenol red is added as pH indicator in the medium.
There are two types of media used for culture of animal cells and tissues
33.4.3. Advantages of serum in culture medium:-
1. Binds and neutralizes toxins.
2. Contains a complete set of essential growth factors, hormones, attachment and spreading factors, binding and transport proteins.
3. Contains the protease inhibitors.
4. Increases the buffering capacity.
5. Provides trace elements.
Disadvantages of serum in culture medium:-
1. It is not chemically defined and therefore it's composition varies a lot.
2. It is sometimes a source of contamination by viruses, mycoplasma, prions etc.
3. It increases the difficulties and cost of downstream processing.
4. It is the most expensive component of the culture medium.
33.4.4. Applications of Animal Cell Culture:-
(a) Production of antiviral vaccines, which requires the standardization of cell lines for the multiplication and assay of viruses.
(b) Cancer research, which requires the study of uncontrolled cell divisions in cultures.
(c) Cell fusion techniques.
(d) Genetic manipulation is easy to carry out in cells or organ cultures.
(e) Production of monoclonal antibodies requires cell lines in culture.
(f) Production of pharmaceutical dregs using cell lines.
(g) Chromosomal analysis of cells derived from the womb.
(h) Study of the effects of toxins and pollutants using cell lines.
- TOOL AND TECHNOLOGY
- HYBRID PLASMID / PHAGE VECTORS
- ARTIFICIAL CHROMOSOMES
- SHUTTLE VECTORS
- ENZYMES USED FOR RECOMBINANT DNA TECHNOLOGY
- DNA LIBRARY
- FLUROSCENT ACTIVATED CELL SORTER
- DNA MICROARRAY OR GENE CHIP OR BIO CHIP
- ANTIBODY GENERATION
- RADIOIMMUNOASSAY (RIA)
- ELISA OR ENZYME LINKED IMMUNOSORBANT ASSAY
- POLYMERASE CHAIN REACTION
- TYPE OF HYDROLYSIS PROBE
- X-RAY DIFFRACTION
- NMR (NUCLEAR MAGNETIC RESONANCE)
- CIRCULAR DICHROISM
- DNA SEQUENCING
- TRANSGENIC ANIMALS
- CRE–LOX P RECOMBINANT SYSTEM
- GENE THERAPY
- TRANSGENIC PLANTS
- PLANT TISSUE CULTURE (PTC)
- MICRO PROPAGATION
- ARTIFICIAL SEEDS
- PRACTICAL APPLICATIONS OF PLANT TISSUE CULTURE
- ANIMAL CELL CULTURE