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23 Cards in this Set
- Front
- Back
name three reasons why biologists have established many cell lines from primary tissues
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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 |
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name two common human cell lines
name other mammalian cell lines |
Hela cervical cancer cells
MCF7 breast cancer cells RAW, CHO, eclone, 3t3 |
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what are J774 cells
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J774. A murine macrophages cell line established from a tumor that arose in a female BALB/c mouse
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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 |
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Before cells lines reach confluency they should by ____ in order to prevent the cultures from dying
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sub cultured
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to subculture cells they need to be brought into ____.
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suspension
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what are proteases used for?
Name one commonly used |
are used to release the cells from the flask
trypsin |
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_____ can hydrolyse the cell adhesion proteins and thus detach the cells from the surface of the flask.
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proteases
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Many cell adhesion proteins (e.g. _____) require _____&_____ to function thus _____ is used when resuspending the cells
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lectins
calcium and magnesium calcium and magnesium free buffer |
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when fresh medium is added to cell suspension the serum in the medium provide two functions..what are they
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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 |
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what are the five physiological parameters for cells?
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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) |
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what are 8 medium requirements?
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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) |
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name 4 issues that arise when cells grow to fill the available area or volume
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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 |
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why is the mass culture of animal cell lines important ?
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to manufacture viral vaccines and many products of biotechnology
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name four biological produced produced by recombinant DNA technology in animal cell cultures
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enzymes
synthetic hormones immunobiologicals (monoclonal antibodies, interleukins, lymphokines) anticancer agents |
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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 |
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what is an example of a complex protein produced in rDNA technology ?
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hormone erythropoietin
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what laminar flow hood do we use and why
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horizontal hoods because they are the best protection for cultures
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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 |
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what type of microscopes are used for visualizing the cells
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inverted phase contrast microscopes
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what cell lines do we use
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J774 eclone macrophage cells
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what are the six general steps of the procedure for splitting cells ?
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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 |
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phenol red (a ph indication) is a constituent of the medium whats it for
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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
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