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23 Cards in this Set

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name three reasons why biologists have established many cell lines from primary tissues
1. to explore the structure and function of cells
2. to investigate the macromolecules important for cellular physiology
3. to study pathways and manifestation of severe disease like cancer, diabetes or even bacterial invasion
name two common human cell lines

name other mammalian cell lines
Hela cervical cancer cells
MCF7 breast cancer cells

RAW, CHO, eclone, 3t3
what are J774 cells
J774. A murine macrophages cell line established from a tumor that arose in a female BALB/c mouse
cell lines are adherent cells that will grow in vitro until when?
two possible answers
they have covered the surface area available (confluency)

or the medium is depleted of nutrients
Before cells lines reach confluency they should by ____ in order to prevent the cultures from dying
sub cultured
to subculture cells they need to be brought into ____.
suspension
what are proteases used for?
Name one commonly used
are used to release the cells from the flask

trypsin
_____ can hydrolyse the cell adhesion proteins and thus detach the cells from the surface of the flask.
proteases
Many cell adhesion proteins (e.g. _____) require _____&_____ to function thus _____ is used when resuspending the cells
lectins
calcium and magnesium
calcium and magnesium free buffer
when fresh medium is added to cell suspension the serum in the medium provide two functions..what are they
1. will deactivate trypsin and allow cells to adhere to the bottom of the flask again
2. provides nutrients and growth factors needed for cell growth
what are the five physiological parameters for cells?
1. temp 37C
2. ph 7.2-7.5
3. humidity
4. gas phase bicarbonate conc. and COs tension in equilibrium (5%)
5.cells should be cultured in the dark (light can have adverse effect on cells)
what are 8 medium requirements?
1 bulk ions: Na K Ca Mg Cl P
2 trace elements iron and zinc
3 glucose (sugar)
4 amino acids
5 vitamins
6 choline, inositol
7 serum
8 antibodies (often used to control growth of contaminants)
name 4 issues that arise when cells grow to fill the available area or volume
1 nutrient depletion in the growth medium
2 accumulation of apoptotic and necrotic (dead) cells
3 cell to cell contact can stimulate the cell cycle to arrest, causing cells to stop dividing know as contact inhibition or senescence
4 cell to cell contact can stimulate cellular differentiation
why is the mass culture of animal cell lines important ?
to manufacture viral vaccines and many products of biotechnology
name four biological produced produced by recombinant DNA technology in animal cell cultures
enzymes
synthetic hormones
immunobiologicals (monoclonal antibodies, interleukins, lymphokines)
anticancer agents
using rDNA more complex proteins that are glycosylated (carbohydrate modified) currently must be made in bacterical cultures

true or false
false they must be made in animal cells

only simpler proteins can be produced using rDNA in bacterial cultures
what is an example of a complex protein produced in rDNA technology ?
hormone erythropoietin
what laminar flow hood do we use and why
horizontal hoods because they are the best protection for cultures
both laminar flow hoods have continuous displacement of air that passes through a HEPA (high efficiency particle) filter that does what?

the hoods are equipped with a short wave UV light that can be turned on for a few minutes to .....
removes particles from the air

sterilize the surfaces of the hood
what type of microscopes are used for visualizing the cells
inverted phase contrast microscopes
what cell lines do we use
J774 eclone macrophage cells
what are the six general steps of the procedure for splitting cells ?
1. view culture in inverted scope to assess degree of confluency
2. remove medium
3. add fresh medium
4. dislodge the cells from the flask
5. transfer 1 ml of cell suspension to new flask containing medium
6. incubate
phenol red (a ph indication) is a constituent of the medium whats it for
as acid by products build up in the indicator will turn from red to orange then yellow. if cultures media turn orange it is probably past due to be fed