Test 2. Lab 6 Flashcards
define cell culture
Refers to the removal of cells from an animal or plant and their subsequent growth in a favorable artificial environment.
The cells may be removed from the tissue directly and disaggregated by enzymatic or mechanical means before cultivation, or they may be derived from a cell line or cell strain that has already been established.
adherent cells
are anchorage-dependent and propagate as a monolayer attached to the cell culture vessel. This attachment is essential for proliferation (fibroblast, astrocytes)
Suspension cells
can survive and proliferate without being attached to a substratum. For example Hematopoietic cells (derived from blood, spleen, or bone marrow) as well as some transformed cell lines and cells derived from malignant tumors can be grown in suspension.
finite cell line
–These are that cells usually divide only a limited number of times before losing their ability to proliferate
continuous cell line
These cell lines are immortal which can occur spontaneously or can be chemically or virally induced.
What are some applications of cell culture
- Excellent model systems for studying the normal physiology and biochemistry of cells such as metabolic studies, aging, the effects of drugs and toxic compounds on the cells, mutagenesis and carcinogenesis.
- It is also used in drug screening and development
- Large scale manufacturing of biological compounds such as vaccines, therapeutic proteins.
*What are some basic requirements for cell culture? Name all and define 4
Cell culture hood (i.e., laminar-flow hood or biosafety cabinet): used to reduce the presence of airborne contaminants
I
ncubator (humid CO2 incubator recommended) : required to maintain optimal carbon dioxide concentration and temperature. For mammalian cells, this is going to be about 5% CO2, the concentration found in the blood and 370C, the normal core body temperature
Water bath: a deviceusedin the laboratories to incubate samples inwatermaintained at a constant temperature. For cell culture temperature of water bath is maintained at 370C.
Centrifuge: used for the separation of fluids, gas or liquid, based on density. Separation is achieved by spinning a vessel containing material at high speed
Refrigerator and freezer: for storage of media and reagents in cold temperature
Microscope: for cell counts using a haemocytometer
Sterilizer (i.e., autoclave): To sterilize equipments
Vessels (flasks, petri dishes, etc) : vessels for growing the cells.
Pipette: For transferring cells, medium etc
*What is the difference between laminar flow and biosafety cabinet?
BSC is mroe versatile because it protects the user and the product in the cabniet from bacterial contamination, can be used for cell cultures materials or drug formations.
The laminar flow only protects the product in the hood. It discharges HEPA-filtered air across the work surface and towards the user. Not ideal for cell cultures
*what are some advantages of Laminar hood vs biological safety cabniet
BSC, user is also protected. It can be used for cell culture techniques.
Laminar flow, sample can still be protected, it is lower cost, can be easier to relocate the BSC
*what is used in the hoods to help remove contaminants from the air before it enters the hood?
HEPA filter (high efficiency particulate air)
*what does typical cell culture media contain?
salts,
amino acids,
sugar,
vitamins
serum - contains a large number of growth promoting activities, buffers toxic nutrients by binding them, neutralizes trypsin and other proteases, contains peptide hormones or hormone-like growth factors that promote healthy growth.
antibiotics - although not required for cell growth, antibiotics are often used to control the growth of bacterial and fungal contaminants.
*What is the most commoly used medium for mammalian culture? and describe
DMEM
Contains approximately four times as much of thevitaminsandamino acidspresent in the original formula and two to four times as muchglucose. Additionally, it containsironandphenol red. DMEM is suitable for most types of cells includinghuman,monkey,hamster,rat,mouse,chickenandfish cells.
What is a astrocyte cell?
A star-shaped glial cell of the central nervous system.
Functions as biochemical support ofendothelial cellsthat form theblood-barrier, provides nutrients to the nervous tissue, maintains extracellular ion balance, repairs and scars brain and spinal cord following traumatic injuries.
- Describe aseptic hood technique
Always wear gloves.
Anything that goes in the hood needs to be sprayed with 70% ethanol (including gloves)
While working, make sure your hands are completely
inside the hood.
- What are the differences between sterile, aspetic and sanitary?
Sterile
Without any microbial contamination
Aseptic
Without pathogenic microbial contamination
Sanitary
Reduced levels PATHOGENIC of microbial contamination to prevent spread of disease.
