Analyzing Cells, Molecules & Systems Flashcards
cell culture
removal of cells from an organism, and promoting their subsequent growth in a favorable artificial environment
types of cell cultures
- primary cell culture
2. established or continuous cell lines
primary cell culture
- taken directly from animal
- involves enzymatic/mechanical selections steps to isolate cells from population
- cells survive for finite amount of time
examples of primary cell cultures
primary neurons
cardiomyocytes
established/continuous cell lines
- a primary culture that has been made immortal by transformation
most common established cell lines
- tumor derived
2. transformed via virus
example of continuous cell lines
- SH-SY-5Y — human neuroblastoma derived
cryopreservation
used to freeze cells
store in liquid nitrogen
advantages of cell culture
- study cell behavior w/out complexity that occur in animal
- cell characteristics can be maintained
- control growth for uniformity
- can expose to agents to see what happens (study drugs, chemicals, etc.)
sub-culture
growing cells and dividing into new
disadvantages of cell culture
- must develop techniques to maintain healthy cells
- quantity of material is limited
- dedifferentiation can occur - which impacts the biology of your cells – making them different than the original cells
applications of cell culture
- research
- disease simulation
- drug testing
- genetic analysis
- produce biological products (hormones, proteins)
- regenerative medicine
cell model of Parkinson’s disease
- expose SH-SY5Y cells to 6-OHDA
- Parkinson’s induced
- creates a reactive oxygen species
- apoptosis triggered
______ helps us to study the unique structure and _____ of individual proteins.
purification
structure and function
purification may use _______ to overexpress a protein - then purify it.
recombinant DNA technology
purification can target ______ proteins as well
endogenous - diseased
sub-cellular fractionation
- -removes all other types of cells
- -reduces the complexity of the material
- -followed by purification
sub-cellular fractionation steps
- tissue sample - mechanical blending
- get homogenate
- centrifuge to separate cell types
- lysis of cells
- ultracentrifugation - separates organelles
homogenate
suspension of different cell types mixed
lysis of cells: sub-cellular fractionation
clean up the cells via
osmotic shock
ultrasonic vibration
mechanical blending
types of ultracentrifuge
fixed angle rotor
swinging bucket rotor–tubes are not rotating at a fixed angle
microsomes
when the ER breaks into little vesicles
when do you use the swinging bucket rotor?
for density gradient centrifugation
column chromatography
apply solvent continuously to top of column and collect fractionated molecules from bottom
