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Semester 1. Practicals > Cell Culture > Flashcards

Cell Culture Flashcards

1
Q

What is cell culture ?

A

Removal of cells from animal or plant and their subsequent growth in a favourable artificail environment

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2
Q

What are the two types of cell culture ?

A
  1. Primary cells
  2. Cell lines
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3
Q

What are primary cells ?

A

Recently isolated from tissues or organs and have mimited lifespan in culture

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4
Q

What are cell lines ?

A

Transformed or immortalised

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5
Q

What are cell cultures a major research tool in ?

A

Cell biology, virology, cancer biotechnology

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6
Q

Who pioneered cell culture and when ?

A

Ross Harrison in 1907

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7
Q

What is the first transformed cell line ?

A

HeLa

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8
Q

What where the HeLa cells used for ?

A

Develop polio vaccine

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9
Q

What are HaLa cells used in ?

A

Cancer, AIDS, effects of radiation and toxic substances, genome mapping

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10
Q

What are the advantages of the primary cells ?

A

Better representation of cells in vivo

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11
Q

What are the disadvantages of primary cells ?

A

Difficulties of isolation, finite life, small numbers of cell obtained, slow growth rate

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12
Q

What are the advantages of cell lines ?

A

Grow continuoudly, easy to handle, large numbers, homogenous constitution, rapid growth rate

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13
Q

What are the disadvantages of cell lines ?

A

Loss of characteristics asociated with original sample, progressive changes in chromosome number

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14
Q

What are the two culturing cell lines ?

A
  1. Suspension cells
  2. Adherent cells
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15
Q

What are suspension cells ?

A
  1. Do not attach to culture vessel
  2. Derived from immune system
    3.To subculture, dilute cells with fresh growth medium
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16
Q

What are adherent cells ?

A
  1. Surface proteins that adhere to culture vessels
  2. To subculture use trypsin/EDTA to detach cells and dilute cells with fresh growth medium
17
Q

What is the source of serum ?

A

Foetal bovine or horse serum

18
Q

What are the disadvantages of serum ?

A
  1. Not the phsiological fluid cells are in contact with in tissue
  2. Protein interferance in bioassays
  3. Contains serum proteases
  4. Constitution not clearly defined
19
Q

What is cryopreservation ?

A

Storage of cells at -180 degrees in liquid nitrogen

20
Q

What does cryopreservation allow for ?

A

Establishment of cell repositories

21
Q

When should cells be cryopreserved ?

A

Exponential growth phase

22
Q

What should you suspend cells in in cyrotherapy ?

A

Glycerol or DMSO (10%) and 90% serum

23
Q

What is the rate of cooling of cryotherapy ?

A

1 degree C/min

24
Q

How do you prepare primary cells ?

A
  1. Isolate tissue
  2. Separate cell from tissue
  3. Incubation and growth
  4. Separate cells from other tissue components
25
Q

How can you separate cells from other tissue components ?

A
  1. Explant culture
  2. Mechanical
  3. Enzyme dissociation
26
Q

What are the applications of cell cultures as model system ?

A
  1. Cell to cell communication
  2. Cellular differentiation
  3. Cell cycle control
  4. Phsiology or biochemistry of cells
  5. Cellular response to invading pathogens
27
Q

What are the applications of cell culture ?

A
  1. Model systems
  2. Drug screening and development
  3. Pathological studies
  4. Virology
  5. Biotechnology
  6. Stem cell research
  7. Tissue engineering
  8. Gene therapy
28
Q

How can cell cultures be used in drug screening ?

A

1.Identification of potentially active compounds
2. Mechanism by which compounds exert their cytotoxic effect
3. Identification of target cell population
4. Identification of toxic concentration range
5. Relationship of drug concentration to exposure time

29
Q

What is transfection ?

A

The process of artifically introducing nucleic acids into cells

30
Q

What are the two types of tranfection ?

A
  1. Transiently
  2. Stable
31
Q

What are the uses of transfection ?

A
  1. Expression of protein of interest
  2. Bioproduction
  3. Stem cell generation
  4. Gene inhibition
32
Q

What are the methods used for transfection ?

A
  1. Chemical gene delivery
  2. Biological gene delivery
  3. Physical gene delivery
33
Q

What are the reasons behind choice of transfection method ?

A
  1. Dependent on cell type and experimental needs
  2. High transfection efficiency
  3. Low cell toxicity
  4. Minimal effects on normal physiology
  5. Easy to use and reproducible

Semester 1. Practicals (2 decks)

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