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BIOL 3321 - cell and molecular bio > culturing cells > Flashcards

culturing cells Flashcards

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1
Q

what did robert hooke do

A

he discovered and named cells

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2
Q

what did antonie van leeuwenkoek do

A

he was the first person to observe and describe living cells

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3
Q

who came up with two of the three tenets that make up cell theory

A

mattias schleiden and theodar schwann

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4
Q

what two tenets did schleiden and schwann come up with

A

living organisms are made up of one or more cells, cell is the basic unit of life

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5
Q

what did rudolph virchow do

A

showed that all cells come from pre-existing cells

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6
Q

define culturing

A

refers to the process of growing cells in an environment that promotes their survival

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7
Q

why are cells cultured

A

to study the properties of them + we use cells to produce lots of products (ie antibodies)

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8
Q

what is 2D culturing

A

cells cultured on a flat surface

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9
Q

what is 3D culturing

A

makes use of a hydrogel, where cells are growing on a 3D scaffold

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10
Q

what two types of cell cultures are there

A

primary cell cultures + cell lines

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11
Q

what are primary cell cultures prepared from

A

animal tissues (ie skin, kidney, liver)

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12
Q

how are primary cell cultures prepared

A

individual cells are prepared by treatment with proteolytic enzymes and then put into the appropriate medium (trypsin+collegenase are used to break the cells away from each other)

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13
Q

T or F: primary cell cultures are the most similar to cells found in an intact organism

A

true

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14
Q

T or F: cells in a primary cell culture will keep dividing

A

false; they will stop after a certain point

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15
Q

after how many divisions in a primary cell culture will cells stop dividing

A

30-50

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16
Q

define cell senescence

A

when cells stop dividing

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17
Q

why do cells in a culture stop dividing after 30-50 divisions

A

production of telomerase turns off, and this is an enzyme that allows cell division to keep happening by keeping the chromosomes at a certain length. without telomerase, chromosomes will get shorter and shorter

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18
Q

define culture shock

A

a protective mechanism that cells have in place when cultured for too long – it makes them stop dividing

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19
Q

define cell lines

A

cell types that will grow indefinitely in culture

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20
Q

why do cell lines grow indefinitely

A

they don’t turn off telomerase production

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21
Q

give an example of cell types that grow indefinitely

A

rodent cell types

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22
Q

why do some rodent cell types grow indefinitely

A

they have a mutation that prevents culture shock from occuring

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23
Q

how can we generate cell lines from other organisms (other than rodents)

A

by using cells from within a tumor

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24
Q

define transformed cell line

A

cells from tumors

25
Q

describe characteristics of transformed cell lines (ie cell lines from the cells of a tumor)

A

transformed cell lines only slightly resemble the parent cell/tissue, they may have altered gene expression, and may have an abnormal karyotype. They can often grow without attaching to a surface, and they can grow to a high density

26
Q

what is the first human cell line

A

HeLa cells

27
Q

where did HeLa cells come from

A

henrietta lacks, who died of cervical cancer. Her cells were removed and distributed without her permission

28
Q

what medical discoveries did HeLa cells help with

A

used to test the polio vaccine, used to show HPV causes cancer, and used in testing vaccines for covid

29
Q

define serum

A

the liquid portion of blood (ie the plasma) where the cells have been removed and the proteins involved in clotting have been removed

30
Q

why is serum used to grow a culture

A

it’s very nutritious; contains amino acids, proteins, vitamins, CHOs, lipids, hormones, growth factors, minerals

31
Q

what is the optimal pH range for mammalian cells

A

7.2-7.4

32
Q

define osmolality + osmole

A

osmolality is the number of osmoles of a solute in a kg of solvent. An osmole is the amount of moles of solute that contribute to the osmotic pressure of a solution

33
Q

what system is used to maintain the pH level

A

CO2/sodium bicarbonate buffering system

34
Q

describe the CO2/sodium bicarbonate buffering system in media

A

as cells grow and respire, CO2 is produced. this dissolved and combined with water to form carbonic acid. This forms H+, causing the pH to decrease. Adding sodium bicarbonate to the medium minimizes the pH drop

35
Q

how much sodium bicarbonate do we add to a media to maintain pH (and how much CO2 is there)

A

1.5g/L of sodium bicarbonate with 5% CO2

36
Q

why is phenol red incorporated into medium

A

it serves as a pH indicator

37
Q

explain how phenol red serves as a pH indicator

A

acidic=yellow, basic=red

38
Q

T or F: cell cultures are grown in an open system

A

true

39
Q

define an open system for growing cultures

A

flask is open to the environment so has exchange can occur, but we use a vented cap that small enough that microbes cannot get in

40
Q

how small are the vents on a vented cap of an open system (what is the purpose of this)

A

0.2um, small enough that microbes cannot get in

41
Q

what are adherent cells

A

anchorage-dependent

42
Q

define confluency

A

the percentage of the surface of the culture dish that is covered by adherent cells

43
Q

at what percentage of confluency do we subculture adherent cells

A

75-90

44
Q

what do we sometimes treat the surface of the culture dish with to benefit the adherent cells

A

treated with something they can adhere to: collagen, laminin, gelatin, fibronectin

45
Q

describe how we subculture adherent cells

A

treat them with protease to lift the cells off the substrate and away from each other (trypsin/EDTA is frequently used)

46
Q

why is EDTA commonly used when subculturing adherent cells

A

because EDTA binds to Ca2+ which is involved with cell adhesion, so this makes it easier to remove the cells

47
Q

what are suspension cells

A

anchorage-independent

48
Q

T or F: suspension cells are easier to subculture than adherent cells

A

true; because we don’t need to use trypsin

49
Q

how do we visualize suspension cells

A

bright field microscope or phase-contrast microscope

50
Q

what two orientations will a microscope have

A

upright or inverted

51
Q

benefit to inverted microscope?

A

objective lenses are below the specimen, so we can look at larger objects and it’s easier/faster to focus (and no risk to the objective lenses)

52
Q

which microscope is inverted

A

phase-contrast

53
Q

which microscope is upright

A

bright-field

54
Q

which cell line are we using in the lab

A

L929

55
Q

where do L929 cells come from (what organism)

A

mice

56
Q

how old are the L929 cells that we’re using

A

100 days

57
Q

what is the morphology of the L929 cells

A

fibroblasts

58
Q

are l929 cells adherent or suspension cells

A

adherent

BIOL 3321 - cell and molecular bio (19 decks)

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