Cell culture Flashcards
1
Q
Primary cells
A
- simple to make but more time-consuming that continuous cell culture
- has normal chromosome number and shape
- require higher conc of growth factors
- many different cell types (mixed) including non-dividing cells (neurons)
2
Q
When will primary cultured cells be essential?
A
For viruses that have advantage living in neurons (e.g. neurons)
3
Q
Continous cell lines made by
A
- isolating cells from tumous
- viral transformation
- oncogene transformation
- mutagenesis
- passaging (mouse and rodent)
- can be cultured indefinitely, but many effects from continuous passaging
4
Q
Applications of cell culture
A
- investigate normal physiology or biochemistry of cells
- test effect of various chemical compounds or drugs
- synthesis of biological materials e.g. proteins
- virus propagation
5
Q
Virus propagation applications
A
- Diagnostic investigation and tests
- measuring infectivity
- evaluation of anti-viral therapies
- Vaccine production
- Basic research
6/ Advanced biotechnology
6
Q
Can virus be grown without animal host or cell line?
A
- bacteriophage are grown in bacteria
- plant viruses grown in plant
- virophage
7
Q
Cell culture medium requirements
A
- balanced salts (isotonic)
- pH 7.2-7.4, buffered w sodium bicarbonate
- pH indicator
- amino acids
- carbs
- foetal bovine serum
- ANTIBIOTICS OPTIONAL
8
Q
Cell culture requirements
A
- culture flasks
- cell culture medium
- incubator
- passage
- freedom from infection
- storage
9
Q
Plaque assays can be used to
A
- determine conc of virus
- determine attenuation of virus stocks
- determine aspects of cytopathic effect
- generate/select recombinant viruses
10
Q
Plaque assay method
A
- Monolayer of adherent cells are grown to merge
- Cells are infected w dilutions of virus stock
- monolayer is overlaid w substance to inhibit diffusion
- Following incubation, live cells are stained w crystal violet
