Virus cultivation and purification Flashcards

1
Q

Most viruses can be grown in

A
  • cultured cells
  • embryonated hen’s egg
  • laboratory animals
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2
Q

Cell culture

A

involves the growth of dispersed animal cells in vitro, either as cells in suspension or as a monolayer on a solid surface

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3
Q

primary cell cultures

A
  • cultures established from tissue taken directly from humans or animals
  • contain several differentiated cell types, which is why they are the best cell culture systems
  • used in producing viral vaccines
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4
Q

Secondary cultures

A

subcultures from primary cultures

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5
Q

serially propagated cell cultures (cell lines)

A
  • diploid cell lines
  • consist of a homogenous population of a single cell type derived from human embryos or subcultures of a primary culture
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6
Q

continuous (Heteroploid) cell lines

A
  • cells are single type that are capable of indefinite propagation in vitro
  • derived from cancer cells
  • their use is prohibited in vaccine production
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7
Q

Trypsinization

A

the organ/tissue is minced and treated with a protease( trypsin or collagenase) to separate the cells

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8
Q

Growth medium

A
  • a chemically defined medium containing all the nutrients required for cell growth, including amino acids, vitamins, inorganic salts and other growth factors such as glucose
  • serum 5-10%
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9
Q

Maintenance medium

A
  • contains 1-2% serum

- used when cell growth is complete

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10
Q

pH indicator

A
  • phenol red

- most media are buffered to pH 7.4

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11
Q

antimicrobial agents

A

penicillin, streptomycin, gentamicin, mycostatin, and kanamycin

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12
Q

Shell vial (1-dram) cell culture

A
  • a small borosilicate glass vial containing a coverslip. the monolayer is grown on the coverslip. it is ideal for centrifuge- enhanced virus inoculation of the monolayer
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13
Q

Co-cultivated cells

A

-this technique involves combinations of different cell types grown together as a single monolayer

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14
Q

yolk sac

A
  • 5-7 day old embryo

- 22 gauge, 1 1/ in length needle

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15
Q

Chorioallantoic membrane

A
  • 9-12 day old embryo

- 26-28 gauge needle

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16
Q

Amniotic route and Allantoic route

A
  • 7-13 day old embryo
17
Q

Intravenous route

A
  • 10- 12 day old embryos
18
Q

Some responses which may be noticed following inoculation of the embryonating egg are:

A
  • stunted growth
  • urate deposits in mesonephros
  • hemorrhage and congestion
  • sluggish movement
  • death of embryo
  • lesions of the extracellular membranes
  • hemaggulutinins in embryonic fluids
19
Q

high-speed equilibrium centrifugation

A

-used to obtain highly purified and con concentrated preparations of viruses

20
Q

buoyant density

A

density at which a virus neither sinks nor floats when suspended in a density gradient