Cell Culture

Question 1

refers to the removal of cells from an animal or plant and their subsequent growth in a favorable artificial environment.

Answer 1

Cell Culture

Question 2

Culture stage after isolation of cells from tissue and proliferated under appropriate conditions.

Answer 2

Primary culture

Question 3

Derived from primary culture.
Genotypic and phenotypic uniformity.

Answer 3

Cell Line

Question 4

Positively selected subpopulation of a cell line done by cloning or other methods.
Acquires additional genetic changes.

Answer 4

Cell strain

Question 5

divide only a limited number of times before losing their ability to proliferate (senescence)

Answer 5

Finite cell lines

Question 6

some cell lines become immortal through a process called transformation

when a finite cell line undergoes transformation and acquires the ability to divide indefinitely

Answer 6

Continous cell lines

Question 7

Cell types based on morphology

Answer 7

• Fibroblast
• epithelial
• Lymphoblast
• Endothelial
• Neuronal

Question 8

Monolayer attached to an artificial
substrate

Appropriate for most cell types, including primary cultures

Answer 8

Adherent cell cultures

Question 9

Free-floating in the culture medium

Appropriate for cells adapted to suspension culture and a few other cell lines that are non-adhesive

Answer 9

Suspension cell cultures

Question 10

Requires periodic passaging

Cells are dissociated enzymatically or
mechanically

Answer 10

Adherent cell cultures

Question 11

Easier to passage, requires daily cell
counts and viability determination

Does not require enzymatic or mechanical dissociation

Answer 11

Suspension cell cultures

Question 12

Growth is limited by surface area

Requires tissue-cultured treated vessel

Used for cytology, harvesting products continuously, etc.

Answer 12

Adherent cell culture

Question 13

Growth is limited to concentration of cells in medium

Can be maintained in vessels not tissue-culture treated

Used for bulk protein production, batch harvesting, etc

Answer 13

Suspension cell culture

Question 14

Cell line selection

Answer 14

• Species
• Finite or Continuos cell line
• Normal or transformed cells
• Growth Characteristics
• Stability
• Phenotypic expression

Question 15

cell culture conditions

Answer 15

• Essential nutrients
• Growth factors
• Hormones
• Gases
• Regulated Physico-chemical environment

Question 16

Basic cell culture equipments

Answer 16

• BSC
• Incubator
• Optical Microscopes
• Water bath
• Centrifuge
• Ultra Low Freezer
• Autoclave
• Cell counter
• Refrigerator

Question 17

Basic cell culture supplies

Answer 17

• Cell culture flasks and well plates
• Micropipettes
• Electric pipette controller
• Serological Pipettes
• Media and reagents
• Centrifuge tubes
• Centrifuge Tube racks

Question 18

Biosafety level for cell culture

Answer 18

BSL 2

Question 19

Biological contaminations

Answer 19

• Bacteria
• Yeast
• Molds
• Virus
• Mycoplasma

Question 20

What is the contaminants

Detected visually
Cloudy culture
pH change

Answer 20

Bacteria

Question 21

What is the contaminant

Detected visually
Clear/turbid culture
No pH change

Answer 21

Yeast and Molds

Question 22

What is the contaminant

Detected by EM, ELISA, PCR, and
immunostaining

Answer 22

Virus

Question 23

What is the contaminant

• Behavior & metabolism
  change
• Detected by PCR,
  ELISA, immunostaining,
  microbioassays,
  autoradiography, etc

Answer 23

Mycoplasma

Question 24

Reputable cell line banks

Answer 24

ATCC or ECACC

Question 25

ATCC

Answer 25

American Type Culture Collection

Question 26

ECACC

Answer 26

European Collection of Authenticated Cell Cultures

Question 27

Applications of Cell culture

Answer 27

- Studying the physiological and biochemistry of cells and organs - Effects of drugs and Toxic compounds - Mutagenesis and carcinogenesis - Drug screening and development - Manufacturing of biological compounds - Tissue engineering and stem cell research

Question 28

5 basic steps of cell cultures

Answer 28

1. PREPARATION OF CELL CULTURE MEDIA 2. THAWING OF CRYOPRESERVED CELL LINE 3. SPLITTING OF CULTURED CELL LINE 4.COLLECTION OF CULTURED CELL LINE 5.CRYOPRESERVATION OF CULTURED CELL LINE

Question 29

Reagents in the cell culture media bottle for 500 mL

Answer 29

- Culture media solution (435 mL) - Fetal Bovine Serum (40mL) - L-glutamine (5 mL) - Penicillin/Streptomycin (5 mL) - Amphotericin (5mL)

Question 30

At what temperature is cell line thawed?

Answer 30

37˚C in water bath

Question 31

WHat will you do after thawing in water bath?

Answer 31

put 10 ml in centrifuge tube and centrifuge at 1500 rpm for 5 mins. Decant the suerpnatant

Question 32

Splitting of Cultures cell line

Answer 32

- Cleaning - Splitting

Question 33

Process of splitting of cultured cell line in cleaning

Answer 33

- Discard old culture media -> add 5ml PBS in culture flask -> Gently mix the flask -> discard the PBS

Question 34

Process of splitting of cultured cell line in Splitting

Answer 34

- Add 2 ml of trypsin in your culture flask -> incubate for 3 min at 37˚C -> view under microscope -> Add 40 mL culture media -> split or transfer 20mL in a new culture flask

Question 35

How to collec culture cell line?

Answer 35

- Disgard old culture media -> perfom cleaning -> add 2mL of trypsin -> for 3 min for 37˚C. -> add 20 mL new culture media -> Collect and transfer in a culture flask/media

Question 36

How to cryopreserve culture cell line

Answer 36

- Centrifuge for 1,500 rpm for 5 min -> Discard supernatant -> Add 5mL freezing media -> Transfer in Cryotubes with labels -> put in box and store at -80˚C

Question 37

where to transfer culture cell line for longer cryopreservation

Answer 37

Liquid nitrogen storage