Unit 1: Cell and Tissue Culture Flashcards
Name examples of where cells can be grown in for cell culturing
in a suspension (liquid medium) or in agar jelly (solid medium)
What conditions are required to ensure rapid growth in culturing?
optimum pH, gases and nutrients
Name specific requirements for cell culturing
Source of cells growth medium type of growth container temperature and pH Gas exchange - supplying oxygen, remove co2 aseptic conditions
what are aseptic conditions?
prevents contamination of media by unwanted microbes and prevents the cell culture itself from becoming contaminated
how are aseptic conditions created?
sterilising with autoclave - heating with steam under pressure
On a graph, what happens during lag phase?
the cells are metabolically active but not dividing
On a graph, what happens during the exponential phase?
cells are dividing at a constant rate
On a graph, what happens during the stationary phase?
due to build up of waste or nutrients becoming limiting, growth stops
On a graph, what happens during death phase?
due to lack of nutrients and build up of waste
Name specific requirements for mammalian cells in culture
growth factors and antibiotics
what are the four stages of subculturing
adhesion, spread, division, confluence
Describe subculturing
Mammalian cells are usually grown in a flask.
cells are detached from the source using proteolytic enzymes such as trypsin.
Cells are added and adhere to the surface of the agar, they flatten out, spread out then start to divide. Cells form a monolayer, one cell thick. When they have completely covered the agar surface - confluent, they stop dividing.
What does pluripotent mean?
able to differentiate into many cell types
what are the differences between microbial and mammalian cultures
Mammalian - complex growth medium, cells divide a finite number of times and once monolayer of cells produced, division stops
Microbial - simple growth medium, essential growth can be maintained and cells are produced until limited by a factor
what are specific culture requirements for plant tissue culture?
M&S salts and growth regulators - auxins and cytokinins
what are the two main methods of plant tissue culture?
explants and protoplasts
what are explants?
small pieces of plant tissue that are placed on a solid medium to promote shoot growth or callus growth.
what are protoplasts?
created by removal of the cell wall with enzyme pectinase and cellulase. result is grown in a medium and treated with growth regulators to make whole new embryonic plant.
why are hybrids so important?
Hybrid plants contain a genetic component from each parental type and therefore desirable traits from both parents
