TIN: Week 4

Question 1

Tissue Culture

Answer 1

Cultivation of eukaryotic tissues outside of the organism, in a growth media with the necessary nutrients, inorganic salts, and pH required to function in a physiological manner

Question 2

Cell Culture

Answer 2

culturing of dissociated cells rather than pieces of tissue

Question 3

Primary Cell Cultures

Answer 3

Primary cell culture is the ex vivo culture of cells freshly obtained from a multicellular organism, as opposed to the culture of immortalized cell lines.

Question 4

Cell Strains

Answer 4

derived from primary cells/ subpopulation of a cell line
has been selected from culture by cloning to possess a specific feature
has a finite lifespan
incapable of growing through serial passaging

Question 5

Cell lines

Answer 5

have the capacity to grow indefinitely
obtained from a primary culture
can grow through serial passaging
the aneuploid population of cells

Question 6

Karyotype

Answer 6

Size, shape, and number of chromosomes in a cell

Question 7

Passaging

Answer 7

detaching cells from the bottom of flask, diluting them, and replating them at a lower density in a new flask
(grown in one environment then removed and put in another)

Question 8

What is tissue culture used for?

Answer 8

• model system for cell biology
• chromosomal disorders
• generation of monoclonal antibodies for production of vaccines as a result of development of hybridoma cell lines
• in vitro fertilization

Question 9

What % CO2 is used to maintain physiological pH in tissue culture incubators?

Answer 9

5%

Question 10

Some cells grow better under lower oxygen conditions? Oxygen can be displaced by what element?

Answer 10

Nitrogen

Question 11

How do you properly freeze cells?

Answer 11

1) Detach using passaging methods
2) suspend in solution with a cryopretectant such as dimethyl sulfoxide (DMSO)
3) Freeze -80 at rate of 1C/min using vessel filled in isopranolol
4) Transfer cells to liquid nitrogen vessel to be stored indefinitely

Question 12

How do you revive cells after being frozen?

Answer 12

Rapidly thaw in 37C water bath

Question 13

In the 1940’s what helped reduce the problem of microbial contamination of cultures?

Answer 13

Penicillin & Streptomycin

Question 14

How do you passage cells?

Answer 14

1) Remove growth media and put on solution containing EDTA
2) Incubate for 5 minutes in EDTA
3) Remove EDTA solution and rinse off with fresh growth media
4) 1/8 slit ratio?

Question 15

How often do iPS (induced Pluripotent Stem) cells need to be re-passaged?

Answer 15

Every 3 Days