In vitro Analyses Flashcards
What is a primary cell?
it’s isolation directly from tissue
morphologically and metabolically similar to parent tissue
can only divide a limited number of times (senescence)
What are immortalized/continuous cells?
mutated cell lines
derived from tumors
morphology and metabolism can differ from parent tissue
What are the seven components of a medium?
Liquid solution is the media Provides nutrients to cells Buffer Environment (temp and concentration of CO2) Phenol Red (pH indicator) Antibiotics and Antimycotics Serum
What is necrosis?
Damage to the cell is severe
In most cases, a rapid injury will result in necrosis
Characterized by ER swelling, membrane blobbing, membrane breakdown, organelle leakage, and random DNA degradation
Often affects groups of cells
Coupled to an inflammatory response
Causes: infection, toxins, trauma
What is apoptosis?
programmed cell death
the cell knows it’s damaged so it will prepare for death
What are the characteristics of apoptosis?
chromatin condensation cell shrinkage preservation of organelles by cell membranes engulfment by neighboring cells membrane blebbing small to no immune response
What are the two apoptotic pathways?
Intrinsic: the cell senses death in itself
Extrinsic: the cell is signaled from other cells to die
What is the caspase?
enzymes playing essential roles in programmed cell death and inflammation.
What is caspase staining?
detects the activity of the enzyme to determine whether or not a cell is undergoing death
What is Anexin V?
used to detect apoptotic cells by its ability to bind to phosphatidylserine, a marker of apoptosis when it is on the outer leaflet of the plasma membrane.
Determined with treating of staurosporine
What is Neutral Red?
a cell viability assay vital dye (vital = able to penetrate living cell) will only stain living cells stains lysosomes taken up by active transport
What is Calcein-AM?
a cell viability assay fluorescent dye taken up by active transport partially metabolized by the cell Calcein-AM in solution out of cell does not glow, converter to calcein and turns green
What is Propidium Iodide?
a cell death assay
intercalating agent (winds itself in DNA)
not membrane permeable (can only enter a broken membrane, hence death)
dye exclusion assay
What is Trypan Blue Exclusion Test?
a cell death assay
vital stain to selectively color dead tissues or cells blue. Live cells or tissues with intact cell membranes are not colored
can not pass through the cell membrane
can bind to serum proteins (not good)
What is CR51 Release?
tests the immune system to kill off cells
used heavily in tumor and viral cytosine studies
Target cells incubated with radio active Cr-51 then exposed to immune cells
If the immune cell kills the target, Cr-51 is released and then measured
