E2 Cell Culturing Flashcards
Used to mimic the occurences in normal basis in live tissue
Primary culture
Cancerous cells that divide indefinitely used in research
Cell lines
True or false, Cell lines are easier to use than primary cultures. Why?
True, Cell lines do not have the Hayflick Limit meaning they are easier to sustain
What are the types of cell lines based on morphology (3)
- Epithelial
- Fibroblastic
- Lymphoblastic
Types of cell lines that attach to the basal lamina
Adherent
Types of cell lines that float in medium
Suspension
4 types of Tissue
ECAM
Epithelial
Connective
Muscular/Skeletal
Neurons
These are cell lines that are tightly packed together
Epithelial
These are cell lines that look like threads
Fibroblastic
Non-confluent and circular cell lines, will move if the plate is disturbed
Lymphoblastic, will move because the cells are in suspension
TIght packing of cells due to several cell divisions
Confluence
Done to avoid confluence and/or to replace medium
Cell Passaging
True or false, both cell lines and primary culture will still divide even under confluency, why or why not?
False, only cell lines as they are unaffected by contact inhibition
This is the eukaryotic equivalent of cell subculturing
Cell passaging
How will you separate cell culture from old medium?
Centrifugation, cells sink into pellet allowing for decanting
4 components of a complete cell culture medium
- Culture Medium
- Buffer system
- Nutrient Serum
- Pen Strep*
- Phenol Red/Indicator**
*Antibiotic (Penicillin) and Antimycotic (Streptomycin)
** Usually omitted but is there to indicate if passage is needed, if contamination is present, etc.
True or false, when using flasks for cell storage, the lids should not be tightly sealed
True, cells need oxygen therefore venting is ncessary
Why are T-flasks lying down during incubation and upright when in use?
Wall mimics the basal lamina for adherent cells that is why it lies down to allow it to settle
Differentiate Biosafety cabinet and Laminar Flow Hood
LFH: Blows air out, Protects your work, but not you
BSC: Airflow is restricted to inside the cabinet, Vertical air flow, protects both the work and worker
How is cell culture contamination determined?
Microscopy (Presence of morphology aside from the 3 previously mentioned)
Color change (Phenol red; rapid change means bacteria, no change after a long time means dead cells; confluence causes acidity meaning yellow)
Used to separate the cells from the T-flask’s simulated basal lamina
Cell dissociation Reagents
3 commonly used CDRs
- Trypsin - Will cut anywhere it can
- Collagenase - Cuts specifically collaged
- Accutase - Used to dissociate nerve cells
