Cell culture Flashcards

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1
Q

What are the 5 universal considerations of cell culture?

A
  1. Aseptic technique - only culture what you choose to culture
  2. Temperature - 37 degrees for mammals, lower for insects, can be very high for archea
  3. Atmosphere - Co2 or O2
  4. Nutrients - solid or liquid medium
  5. Safety
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2
Q

What conditions must Schizosaccharomyces pombe be gown under?

A
  • Agar, however different nutrients than bacteria
  • Atmospheric gases
  • Different temperature
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3
Q

What can yeast be used for?

A

S. pombe - Model for the cell cycle, as have simple eukaryotic genome and similar components and cycle
S. cervisiae - beer and studies of cell trafficking

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4
Q

What conditions must Haloferax be grown in?

A

45 degrees with high salt concentrations, has red colour due to lycopene

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5
Q

How are cell cultures of multicellular organisms usually conducted?

A
  • Liquid culture
  • Usually red, indicating pH of 7.4. Bi-products such as lactic acid and pyloric acid can lower pH and turn indicator yellow
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6
Q

What was Alexis Carrel the first to produce?

A

A cell line of chick embreyonic fibroblasts described as “immortal”, living for a long time under aseptic conditions when fed with nutrients.
However, doubt over whether a malignant transformation was present or if accidental contamination with fresh cells had occurred

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7
Q

How can one produce a primary cell line?

A
  • Isolate cells from tissue by dissection, mincing or enzymatic dissociation
  • Dealing with cells directly from the organisms, although these are frequently short lived and it’s hard to maintain consistency
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8
Q

How can one produce a transformed (immortal) cell line?

A
  • Take cells from a tumor
  • Expression of viral proteins such as Large T-antigen (SV40)
  • Express genes that confer immortality such as telomerase
  • However, not as representative of cells in the body
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9
Q

What is 2D culture?

A

Use of glass/plastic flask where cells adhere to the base and culture on the medium

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10
Q

How is 2(1/2)D culture different from 2D culture?

A

Extracellular matrix at the base of the flask which cells can modify to create “organs”. Also contains different components.

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11
Q

What is 3D culture?

A

Where a 3D agar gel is used to culture stem cells/mammalian cells in order to produce precise patterning

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12
Q

How does mechanically supported culture exploit the air/liquid barrier?

A

Grows organ explant at air/liquid interface to exploit O2 perfusion

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13
Q

Name 3 reasons why cell culture is important

A
  1. In order to reduce, refine and replace animal studies
  2. Scarce availability of human tissues and ethics problems in obtaining them
  3. Usage for regenerative medicine e.g stem cells
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