E2 Cell Culturing

Question 1

Used to mimic the occurences in normal basis in live tissue

Answer 1

Primary culture

Question 2

Cancerous cells that divide indefinitely used in research

Answer 2

Cell lines

Question 3

True or false, Cell lines are easier to use than primary cultures. Why?

Answer 3

True, Cell lines do not have the Hayflick Limit meaning they are easier to sustain

Question 4

What are the types of cell lines based on morphology (3)

Answer 4

1. Epithelial
2. Fibroblastic
3. Lymphoblastic

Question 5

Types of cell lines that attach to the basal lamina

Answer 5

Adherent

Question 6

Types of cell lines that float in medium

Answer 6

Suspension

Question 7

4 types of Tissue

Answer 7

ECAM
Epithelial
Connective
Muscular/Skeletal
Neurons

Question 8

These are cell lines that are tightly packed together

Answer 8

Epithelial

Question 9

These are cell lines that look like threads

Answer 9

Fibroblastic

Question 10

Non-confluent and circular cell lines, will move if the plate is disturbed

Answer 10

Lymphoblastic, will move because the cells are in suspension

Question 11

TIght packing of cells due to several cell divisions

Answer 11

Confluence

Question 12

Done to avoid confluence and/or to replace medium

Answer 12

Cell Passaging

Question 13

True or false, both cell lines and primary culture will still divide even under confluency, why or why not?

Answer 13

False, only cell lines as they are unaffected by contact inhibition

Question 14

This is the eukaryotic equivalent of cell subculturing

Answer 14

Cell passaging

Question 15

How will you separate cell culture from old medium?

Answer 15

Centrifugation, cells sink into pellet allowing for decanting

Question 16

4 components of a complete cell culture medium

Answer 16

1. Culture Medium
2. Buffer system
3. Nutrient Serum
4. Pen Strep*
5. Phenol Red/Indicator**

*Antibiotic (Penicillin) and Antimycotic (Streptomycin)
** Usually omitted but is there to indicate if passage is needed, if contamination is present, etc.

Question 17

True or false, when using flasks for cell storage, the lids should not be tightly sealed

Answer 17

True, cells need oxygen therefore venting is ncessary

Question 18

Why are T-flasks lying down during incubation and upright when in use?

Answer 18

Wall mimics the basal lamina for adherent cells that is why it lies down to allow it to settle

Question 19

Differentiate Biosafety cabinet and Laminar Flow Hood

Answer 19

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

Question 20

How is cell culture contamination determined?

Answer 20

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)

Question 21

Used to separate the cells from the T-flask’s simulated basal lamina

Answer 21

Cell dissociation Reagents

Question 22

3 commonly used CDRs

Answer 22

1. Trypsin - Will cut anywhere it can
2. Collagenase - Cuts specifically collaged
3. Accutase - Used to dissociate nerve cells