Experiment 1 Flashcards
(35 cards)
Tissue culture
Culture of isolated cells. Not just applicable to human or animal cells, many of the same techniques used to grow plant and insetcs in the laboratory as well
Primary Explants
individual cells derived directly from tissues or blood
Continuous cell line
transformed cells or individual cells isolated from a tumor
Types of cell lines
Non-transformed (normal, limited life span)
Transformed (altered to show continual growth)
Cancer cell lines (derived from cancers, show continual growth, may have altered function)
Adherent cells
Attach to culture dishes, require brief enzymatic digestion with a protease (i.e. trypsin) to remove the attached cells from a culture dish
Adherent cell examples
Epithelial, endothelial, neuronal, fibroblasts
Epithelial cell types
Skin, intestinal/colon mucosa, liver or other organs
Usually grow flat and spread out
Fibroblasts
Connective tissue
Grow as spindle shape
Endothelial cells
Line blood vessels
Grow spreading and flat
Cell types we use in experiment 1
3T3: mouse fibroblast, transformed
HT29: human colon carcinoma, cancer
Cell suspension/non-adherent cells
easily pipetted out of the dish with the medium
Non-adherent cell examples
Blood leukocytes
In our lab, we are using (cell lines or primary explant) which will grow as (adherent cells/cell suspension)
cell lines; adherent cells
How do we feed adherent cells to provide them with necessary nutrients? What needs to be done to allow for growth over time?
Remove spent medium from the culture dish and add in new fresh medium. A portion of cells needs to be removed and inoculated into a new dish or flask with fresh medium to allow for growth to continue
Passage
Length of time that the cells have been maintained in culture
What would happen if we simply tried to put our cells into a dish and grow them without making a conscious effort to remove any wayward bacterium?
Bacterium would soon end up mixing in the culture with the cells; leaving few living cells.
Sterile
Microbe free, including bacteria, fungi and virus
What cells did we use in the experiment?
3T3 (mouse fibroblast, transformed)
HT29 (human colon carcinoma, cancer)
What are the 3 methods used to avoid contamination of culture/sterilize growth medium? What is another way that could be considered “cheating”?
- Sterilize solution and equipment
–>Autoclave
–>Filtration
–>Irradiation - Aspectic or sterile technique
- laminar flow hood
- antimicrobials (antibodies) (not used for sterilization)
Filtration
pass solutions through a membrane with a small pore size
Autoclave
uses high pressure and temperature to kill microbe
Irradiation
X or gamma irriation to kill microbes (NO UV)
True or false; antibiotics can be used for sterilization
false; not used for sterilization. Just used to avoid contamination
True or false; we must culture the cells in a appropriate environment complete with all the nutrients, vitamins, hormones, salts, oxygen, pH and temperature to mimic the living body
True
