Cultivation of Viruses Flashcards
T/F: Viruses have the genetic capability to multiply by division?
FALSE, need to use host machinery to make more virions
Methods for growing/cultivating viruses:
Cell tissue culture
Innoculation in embryonated egg
Laboratory animals
A culture in which the bottom of the culture vessel is covered by a continuous, single layer of cells.
Monolayer culture
A culture of cells taken directly from parental tissue (liver, lungs, muscle):
Primary cell culture
Advantages of primary cell cultures:
Best culture system for isolation/propagation of viruses.
Used in producing viral vaccines.
Subcultures, the transfer of cells from one culture vessel to another, is used to:
Provide fresh nutrients and space for growing cultures
After the first subculture, the primary culture is known as a:
Cell line
What are the 2 types of cells lines?
- Finite/diploid (limited life span/limited generations)
2. Continuous (acquire ability to divide infinitely)
Finite/diploid cell lines have a limited life span and are derived from:
Embryos, or from secondary cell cultures
What cell lines can be used for vaccine production?
finite/diploid
Continuous cell lines (immortal/heteroploid cell lines) are derived directly from:
Cancer cells
Can continuous/immortal cell lines be used in vaccine production?
Hell no
What provides necessary nutrients for cell growth in the lab?
Culture media
What can be added to culture media to aid in growth and adhesion, regulate cell membrane permeability, and provide nutrients?
Serum
What is the most common animal serum supplement used in the lab?
FBS - fetal bovine serum
What is used as a pH indicator and changes the medium from red to yellow when the pH drops?
Phenol Red
It is necessary to use exogenous CO2 when using media buffered with a CO2-bicarbonate based buffer to maintain:
pH of the medium
What can be added to culture media to prevent contamination with bacteria, mycoplasma, yeast and molds?
antimicrobial agents
What proteolytic enzyme is used to detach and dissociate cells while subculturing?
Trypsin
What is cytopathic/cytopathogenic effect (CPE)?
Damage or morphological changes to host cells during viral invasion
Types of embryonated egg inoculation
Yolk Sac Inoculation
Allantoic Cavity Inoculation
Amniotic Cavity Inoculation
Chorioallantoic Membrane Inoculation (CAM)
In egg inoculations, what are the main signs of virus growth?
Death of the embryo
Paralysis
Stunted growth
What are the little white foci that form on the chorioallantoic membrane as a result of virus growth?
Pocks
Locations on mice for inoculation of viruses in the lab
intracerebral
intraperitoneal
