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Cell and Tissue Engineering > Quiz 2 Slides (Lecs. 5-7) > Flashcards

Quiz 2 Slides (Lecs. 5-7) Flashcards

1
Q

Blast cells

A

immature, undifferentiated cells that become less active and mature into cytes

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2
Q

Cyte cells

A

mature, differentiated cells that can revert back into blasts

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3
Q

What are the components of cell culture media? (9)

A

Water, Balanced Salts, energy molecules, buffering agents, pH indicators, anti bacterial substances, amino acids, vitamins, growth factors

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4
Q

What is the function of balanced salts in cell culture media?

A

provide a physiologic ionic environment and maintain osmotic balance

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5
Q

What is the function of buffering agents in cell culture media?

A

maintain pH of cells around 7.4

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6
Q

What is the function of phenol red?

A

pH Change indicator

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7
Q

What is the purpose of a biosafety cabinet during cell culture?

A

prevent airborne particles from contaminating cells

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8
Q

What is the purpose of a culture flask?

A

maintain a sterile environment and give cells a substrate to bind to

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9
Q

Why use an inverted microscope to view cells?

A

can look directly at cells instead of looking at humidity and other molecules on the top of the glass slide

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10
Q

What are the 4 common cell culture contaminants?

A

bacteria, yeast, fungi (mold), viruses/mycoplasma

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11
Q

What is the bicarbonate buffer system?

A

acid-base mechanism involving the balance of carbonic acid, bicarbonate, and CO2 in order to maintain pH

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12
Q

What is the most common cell detachment solution?

A

trypsin; an enzyme which breaks down proteins and dissociates adherent cells

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13
Q

What is trypan blue?

A

dye used to differentiated between live and dead cells

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14
Q

What is cell proliferation?

A

process that results in an increase in cell number

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15
Q

What are the pros of in vivo cell culture?

A

native 3D environment and all relevant signals present

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16
Q

What are the cons of in vivo cell culture?

A

many variables leading to noisy data and animal rights concerns

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17
Q

What are the pros of in vitro cell culture?

A

simplified models, observe as function of time, study parameters independently

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18
Q

What are the cons of in vitro cell culture?

A

unnatural 2D enviroment, and may lack important signals

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19
Q

What is an advantage of 3D in vitro cell culture?

A

more accurate representation of in vivo cell growth. and more in vivo like w/o animal rights concerns

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20
Q

What factors affect cell proliferation?

A

temperature, nutrients, waste removal, cell type, culture type and in vitro vs in vivo

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21
Q

primary cultures

A

cells isolated directly from living organism, will eventually die off

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22
Q

Cell lines

A

derived from primary cells, shows exponential growth

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23
Q

Why is the constrained growth model a better approximation of proliferation?

A

takes into account things that constrain cell growth such as contact inhibition

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24
Q

Stem cells

A

undifferentiated cells with self renewal capabilities

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25
Q

reprogrammed cells

A

differentiated cells that have been altered to become less differentiated

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26
Q

What is passaging?

A

removal of the medium and transfer of cells from a previous culture into new culture with fresh medium

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27
Q

What is expansion during passaging?

A

moving cells from one dense flask into a larger flask or multiple flasks

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28
Q

When are cells passaged in an adherent culture?

A

when the cells cover all the available substrate and have no more room to grow

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29
Q

When are cells passaged in a suspension culture?

A

when the cells exceed the nutritional capacity of the medium

30
Q

What is the term P1 cell culture

A

once cells have been re-plated they are termed starting with P1 then P2

31
Q

what is senescene?

A

when cells reach their lifetime limit

32
Q

What is Hayflick limit?

A

the number of doublings that a cell can undergo (typ. 50-70)

33
Q

What are the 3 common cell harvesting methods?

A

bone marrow aspirate, uroepithelial flush out, tissue biopsies

34
Q

Bone marrow aspirate

A

use large needle w/ local anesthesia to withdraw about 2mL of marrow

35
Q

Uroepithelial cell flush out

A

no anesthesia required, repeated filling/emptying w/ isotonic salt solution

36
Q

Tissue biopsies

A

take punch from skin, arthroscopy to harvest, or fluoroscopy and catheter

37
Q

Mechanical disruption

A

vortex w/ digestion buffer, pipet vigorously, dice w/ a scalpel

38
Q

Enzymatic disruption

A

disrupt cell-ecm or cell-cell connections

39
Q

Selective adhesion

A

isolated desired cell types by manipulating adhesion characteristics based on expression of cell surface receptors or adhesion speed

40
Q

Cell selection techniques

A

selective adhesion, differential centrifugation, gradient centrifugation, antibody driven, FACS

41
Q

Differential centrifugation

A

separation of cells entirely based on size and mass

42
Q

isopycnic gradient centrifugation

A

for separation as a function of density only, high density gradient

43
Q

rate zonal gradient centrifugation

A

for separation of particles primarily based on size and shape

44
Q

antibody driven cell selection

A

select a target antigen on cell surface

45
Q

fluorescent activated cell sorting

A

label cells w/ fluorescent antibody and flow through machine to sort

46
Q

adherent cells

A

anchorage dependent and must be cultured while attached to a solid or semi solid substrate

47
Q

suspension cells

A

anchorage independent, can be grown floating in the medium

48
Q

Embryonic stem cells

A

cultured of isolated inner cell mass and/or primordial germ cells

49
Q

embryonic stem cell limitations

A

propensity to form teratomas

50
Q

Somatic cell nuclear transfer

A

take normal oocyte and remove nucleus then replace with nucleus from a somatic cell

51
Q

totipotent stem cells

A

give rise to any of the 220 cell types found in an embryo as well as the placenta

52
Q

pluripotent stem cells

A

give rise to all cell types but not the placental cells

53
Q

multipotent stem cells

A

develop into a limited number of cell types in a particular lineage

54
Q

Induced pluripotent stem cells

A

artificially created stem cells that are forced to express certain genes

55
Q

progenitor cells

A

tendency to differentiate into a specific type of cell but is more specific than a stem cell

56
Q

what is stemness

A

refers to an essential characteristic of a stem cell that distinguishes it from ordinary cells

57
Q

chimera

A

single organism composed of cells from different zygotes

58
Q

pros of using stem cells in tissue engineering

A

stem cells typically reduce and/or eliminate tissue rejection or failure

59
Q

cons of using stem cells in tissue engineering

A

they differentiate too quickly and they can be tumorigenic

60
Q

reprogramming cells

A

goal is to induce differentiated cell to revert back to pluripotency

61
Q

transdifferentiation

A

cells regress to a point where they can switch lineages allowing them to differentiate into another cell type

62
Q

dedifferentiation

A

terminally differentiated cell reverts back into a less differentiated stage

63
Q

What factors influence differentiation?

A

biochemical reagents, changing culture media

64
Q

stem cell niches

A

the microenvironment; specific anatomic location which regulates how they participate in tissue generation/maintenance

65
Q

Cell function assays

A

adhesion, metabolism, migration, proliferation

66
Q

transwell membrane assay

A

used to assess cell migration towards a chemotatic stimulus;

67
Q

scratch assay

A

artificial gap is created on cell monolayer and cells move towards it

68
Q

Laminar flow

A

no mixing between layers; Re < 2100

69
Q

turbulent flow

A

irregular movement of particles; Re > 2400

70
Q

Alamar blue reaction

A

used to assess cell metabolism; add alamar blue, incubate, and measure absorbance

71
Q

Calcein assay

A

measurement of cell viability; cells are incubated in solution and fluorescent cells are imaged

Cell and Tissue Engineering (6 decks)

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