Animal cell cultivation involves growing cells outside their natural environment using defined conditions. Primary cell cultures are obtained directly from tissue and consist of heterogeneous cells that divide a limited number of times. Secondary cultures, or cell lines, are produced by sub-culturing primary cultures and can divide indefinitely. Cells are grown using natural media like serum or artificial chemically-defined media and maintained through aseptic techniques, cryopreservation, and various culture methods. Cell culture has applications in vaccine production, research, and biotechnology.

1. Animal cell cultivation
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2. Cell culture:
• Cell culture is the process by which cells are grown
under controlled conditions, generally outside of their
natural environment .
• Cell culture was first successfully undertaken by Ross
Harrison in 1907.
• Roux in 1885 for the first time maintained embryonic
chick cells in a cell culture.
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3. Animal cell culture:
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• Invitro cultivation of animal
organs ,cells and tissues at
defined temperature using an
incubator and supplemented with
a medium containing cell
nutrients and growth factors is
collectively known as animal cell
culture .

4. Types of animal cell culture:
• Based on the number of cell division, cell culture can be
classified as
- Primary cell culture and
- Secondary cell culture/Cell lines.
• Cell lines can undergo finite or infinite cell divisions.
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5. Primary cell culture:
• The cell culture obtained straight from the cells of a host
tissue.
• The cells dissociated from the parental tissue are grown
on a suitable container and the culture thus obtained is
called primary cell culture.
• Such culture comprises mostly heterogeneous cells and
most of the cells divide only for a limited time.
• However, these cells are much similar to their parents.
• Depending on their origin, primary cells grow either as
an adherent monolayer or in a suspension.
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6. Secondary cell culture :
• When a primary culture is sub-cultured, it is known as
secondary culture or cell line or sub-clone.
• Primary cell culture cannot viable for a long time because
the cell utilize all nutrients of the medium .Therefore the
subculture is needed.
• During repeated sub-culturing and selection ,the cell line
gets evolved and properly established consisting of
proliferating cells.
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7. Culture media:
• Two types of culture media used in animal cell culture.
- Natural media
- Artificial media
Natural media :
• These media consist of naturally occurring biological fluids .
• There are three types:
- Clots
- Biological fluids
- Tissue extracts
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8. Clots:
• The most commonly used clots are plasma clots,which have been in
use for a long time.
• Plasma is now commercially available in liquid state.
Biological fluids :
• Various biological fluids can be used as culture media .
• Example : Serum ,insect hemolymph,amniotic fluid .
• Serum is the most widely used .
Tissue extracts:
• Chick embryo extract is the most commonly used tissue extract.
• Bovine embryo extract is also used.
• Tissue extract can often be substituted by a mixture of amino acid
an certain other organic compounds .
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9. Artificial media:
• Artificial media may be grouped into following types:
- Serum containing media
- Serum free media
- Chemically defined media
- Protein free media
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10. Serum containing media:
• Fetal bovine serum is the most common supplement in
animal cell culture media .
• Serum provides carriers water-insoluble nutrients,
hormones and growth factors ,protease inhibitors.
Serum free media:
• It is referred to as defined culture media .
• These media are generally specifically formulated to
support the culture of single cell type and incorporate
defined quantities of purified growth factors
,lipoproteins and other proteins.
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11. Chemically defined media :
• These media contains pure protein additives .
• It also contains contamination free ultra pure inorganic
and organic ingredients.
• Their constituents are produced in bacteria or yeast by
genetic engineering .
Protein free media:
• It do not contain any protein and only contain non protein
constituents .
• It promotes superior cell growth and protein expression .
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12. Methods of cell cultivation:
Aseptic techniques:
• Bacterial infections like Mycoplasma and fungal infections commonly
commonly occur in cell culture creating a problem to identify and
eliminate.
• All cell culture work is done in a sterile environment with proper
aseptic techniques.
• Work should be done in laminar flow with constant unidirectional
flow of HEPA filtered air over the work area.
• All the material, solutions and the whole atmosphere should be of
contamination-free.
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13. Cryopreservation:
• If a surplus of cells is available from sub-culturing they should be
treated with the appropriate protective agent( Eg: DMSO or glycerol)
glycerol) and stored at temperature below 130ºC until they are
needed.
• This stores cell stocks and prevents original cell from being lost due to
due to unexpected equipment failure or biological contaminations.
contaminations.
• It also prevents finite cells from reaching senescense and minimizes
minimizes risks of changes in long term cultures.
• When thawing the cells, the frozen tube of cells is warmed quickly in
quickly in warm water, rinsed with medium and serum and then
added into culture containers once suspended in the appropriate
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15. Applications:
• Vaccines production
• Cellular and molecular biology
• Virus cultivation
• In cancer research
• Gene therapy
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16. References :
• Bioprocess engg – Michael.L.Shuler and Fikret kargi
• Pictures – google images
• Methods – https://microbeonline.com
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