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Lecture 4

BIOL483- Lecture 4 - Sept 19.docx

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BIOL 483
Vivian Dayeh

Nutrient environment used for cell culture is made from what main components? - Serum - Antibiotics - Basal medium These three components make up complete medium! Important to grow cell, and allow simulation for cells to grow. Antibiotics - Animal cells are much slower in growth when compared to bacterial or yeast cells - Want to eliminate contaminants (bacterial cells) Penicillin in streptomycin is most common combination in addition to basal medium. Not a very strong amount about 2% of antibiotics, just to control any microbes that may grow. Ampho B not as commonly added. Disadvantages - If your culturing technique is good, you don’t need antibiotics (not a disadvantage, but opposite of it) Easiest way to know complete medium - Shake it and it will foam up, serum will FOAM UP. - Basal medium WONT do this Depends on cell, but many cells that are normal tend to grow upon one layer and that’s it. Some cells grow ontop of each other. Polystyrene = most common plastic - Must be treated to have net negative charge on bottom so cells can be attracted to it and create attachment. Multiwell plates - Can do invidividual experiment, have cells that will grow at bottom of each well. - 48-96 wells common. - Specialized plates treated on the bottom for cells to attach Primary cell culture 3 main methods to produce PRIMARY CELL CULTURES. (initiate culture) 1. Explant outgrowth a. Very small piece of tissue (explant) b. Dish that’s open in media (everything must be sterile), snip as much a
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