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Animal Biotech 3
Animal Biotech 3
Animal Biotechnology
Haji Akbar
Establishment of cell culture:
Many types of animal cells can grow in vitro
such as tumour cells, neuroblastoma cells etc.
Ability to translocate,
Factors affecting subculture in vitro:
Several factors that influence differentiation and
proliferation of cells when subculture in vitro as
below;
Application in biotechnology:
Gene mapping
Malignancy
Viral replication
Antibodies production via hybridoma technology
Continue!!!
Macrophages fuse around the foreign body or bacterial
cell in the tissues.
* Lag phase
: adapt to new environment; repair cell membrane
damage
* Log phase
: exponential growth: 90-100% of cells are dividing
Trypsin (crude)
: from cattle and pigs pancrease
: contain Chymotrypsin, elastase, ribonucleas
deoxyribonuclease and amylase
** Enzymes **
Collagenase
: for connective tissue
Pronase
: for fibroblast
Elastase
: for fibroblast protein
Deoxyribonuclease
: for DNA
Preparation of cell suspension from intact tissue
(cont.)
Source of tissue
: Young animals
e.g. kidney (Monkey, Dog, Rabbit),
Chick embryo
: Old animal tissue contains a large
amount
of connective tissue
Preparation of cell suspension from intact tissue
(cont.)
1. Removal of tissue and place in Isotonic
(or growth medium with antibiotic)
subculture
Cell line
2. Continous stirring
fungal contamination
Bacterial contamination
Mycoplasma contamination
Viral contamination
Other cell line
Sources of contamination
Incubator : humidified
Incubation temperature
pH and Buffer
Gas phase
Osmolarity and water grade
Cell Storage
: Freezing Medium
* Serum (~ 20-90%)
* Culture medium
* Cryoprotective agent : ~ 5-10%
(DMSO, Glycerol)
Cell Storage
* (-20oC) Freezer
Epithelium-liked cell
Monolayer cell culture