Animal cell cultivation

Animal cell cultivation

1. Animal cell cultivation Done by, Pavithra .P(117011101298) Pavithra .T(117011101299) Pooja.K(117011101300) 11-05-2020 B.Tech( Biotechnology) 1

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. 11-05-2020 B.Tech( Biotechnology) 2

3. Animal cell culture: 11-05-2020 B.Tech( Biotechnology) 3 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 . 11-05-2020 B.Tech( Biotechnology) 4

5. Primary cell culture: T he 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 . 11-05-2020 B.Tech( Biotechnology) 5

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. 11-05-2020 B.Tech( Biotechnology) 6

7. Culture media: Two types of culture media used in animal cell culture. - Natural media - Artificial media N atural media : These media consist of naturally occurring biological fluids . There are three types: - Clots - Biological fluids - Tissue extracts 11-05-2020 B.Tech( Biotechnology) 7

8. Clots: T he 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 . 11-05-2020 B.Tech( Biotechnology) 8

9. Artificial media: Artificial media may be grouped into following types: - Serum containing media - Serum free media - Chemically defined media - Protein free media 11-05-2020 B.Tech( Biotechnology) 9

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. 11-05-2020 B.Tech( Biotechnology) 10

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 . 11-05-2020 B.Tech( Biotechnology) 11

12. Methods of cell cultivation: Aseptic techniques: Bacterial infections like Mycoplasma and fungal infections 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. 11-05-2020 B.Tech( Biotechnology) 12

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) and stored at temperature below 130ºC until they are needed. This stores cell stocks and prevents original cell from being lost due to unexpected equipment failure or biological contaminations. It also prevents finite cells from reaching senescense and minimizes risks of changes in long term cultures. When thawing the cells, the frozen tube of cells is warmed quickly in warm water , rinsed with medium and serum and then added into culture containers once suspended in the appropriate media. 11-05-2020 B.Tech( Biotechnology) 13

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15. Applications: Vaccines production Cellular and molecular biology Virus cultivation In cancer research Gene therapy 11-05-2020 B.Tech( Biotechnology) 15

16. References : Bioprocess engg – Michael.L.Shuler and F ikret kargi Pictures – google images Methods – https://microbeonline.com 11-05-2020 B.Tech( Biotechnology) 16

17. 11-05-2020 B.Tech( Biotechnology) 17 Thank you