Cultivation of Viruses + Virus Purification & Concentration Flashcards
What are the possible ways to grow and cultivate viruses?
- Cell / Tissue culture
- Inoculation in embryonated egg
- Lab animal inoculation
What are the 2 types of cell culture?
- Suspension culture
- Monolayer culture
What is the mechanism of suspension cultures?
cells dont require attachment for growth + dont attach to surface of the culture
- cells suspended in media / propagated in suspension
What is the mechanism of monolayer cultures?
the bottom of the culture is covered with continuous layers of cells, 1 cell in thickness
What are the advantages of primary cell culture?
- best for isolation + propagation of viruses
- Heterogeneous - many cell types
- Closest to animal / similar to animal tissue cells
- Used to make viral vaccines
What are the disadvantages of primary cell culture?
- Susceptible to contamination
- Complex, may not fully act like parent tissue
- Short lifespan in culture
- Difficult to obtain
What is subculture?
When you transfer cells from primary cell culture to a new cell culture
Ex: primary cell culture to secondary cell culture
Why is subculturing neccessary?
Required to provide fresh nutrients and growing space for continuously growing CELL LINES
What types of cell lines are there?
- Continuous
- Finite/ diploid
What is the mechanism of finite/diploid cell lines?
- limited life span with limited # of cell generation
** can only be subcultured a certain amt of times before it cant anymore
What is the mechanism of continuous cell lines?
Has the ability to divide indefinitely
** can be continuously subcultured
True or False: In rate-zonal centrifugation, virus particles are separated based on their buoyant density
FALSE - based on mass
What ways can be used for purification of viruses?
- Membrane chromatography
- Isopycnic centrifugation
- Rate-zonal centrifugation
____________ plays a vital role in virology by efficiently sedimenting the smallest viruses
Ultracentrifuge
________ separates viral particles on the basis of their buoyant density and aids in purification and concentration of viruses
Isopycnic centrifugation
What is rate-zonal centrifugation?
Ultracentrifugation method for purification and concentration of viruses
- separates virus particles based on their MASS
** bc particles move @ dif rates depending on their mass when under centrifugal force
What is a primary cell culture?
Maintenance of growth of cells dissociated directly from parental tissue (Ex: kidney or liver) of human or animal origin
What are some characteristics of finite/diploid cell lines?
- Limited life span
- Homogenous population of single cell type
- Derived from embryos or secondary cell culture
- *SLOW GROWTH RATE
- Cells retain original morphology
- Cells exhibit contact inhibition, density limitations + anchorage dependance
- *USED FOR VACCINES!
What are the characteristics of immortal/ heteroploid cell lines?
- Mostly homogenous w single cell type
- **DERIVED FROM CANCER CELLS
- **FDA PROHIBITS USE FOR VACCINES
- Rapid growth rate
- No contact inhibition or anchorage dependance
- Genetically, furthest away from the animal
Which cell line can be used for vaccine production?
Finite/ diploid cell lines ONLY
Which cell line exhibits anchorage dependance?
Finite/diploid
What are examples of a continuous cell line?
- HeLa cells
- Henrietta Lacks cells
What morphology can viral cells in culture have?
- Fibroblastic
- Epithelial-like
- Lymphoblast-like
What are the characteristics of a fibroblastic cell in culture?
- bipolar or multipolar
- Elongated shapes
- ** grows attached to a substrate
