Cell Culture

Question 1

What is cell culture ?

Answer 1

Removal of cells from animal or plant and their subsequent growth in a favourable artificail environment

Question 2

What are the two types of cell culture ?

Answer 2

1. Primary cells
2. Cell lines

Question 3

What are primary cells ?

Answer 3

Recently isolated from tissues or organs and have mimited lifespan in culture

Question 4

What are cell lines ?

Answer 4

Transformed or immortalised

Question 5

What are cell cultures a major research tool in ?

Answer 5

Cell biology, virology, cancer biotechnology

Question 6

Who pioneered cell culture and when ?

Answer 6

Ross Harrison in 1907

Question 7

What is the first transformed cell line ?

Answer 7

HeLa

Question 8

What where the HeLa cells used for ?

Answer 8

Develop polio vaccine

Question 9

What are HaLa cells used in ?

Answer 9

Cancer, AIDS, effects of radiation and toxic substances, genome mapping

Question 10

What are the advantages of the primary cells ?

Answer 10

Better representation of cells in vivo

Question 11

What are the disadvantages of primary cells ?

Answer 11

Difficulties of isolation, finite life, small numbers of cell obtained, slow growth rate

Question 12

What are the advantages of cell lines ?

Answer 12

Grow continuoudly, easy to handle, large numbers, homogenous constitution, rapid growth rate

Question 13

What are the disadvantages of cell lines ?

Answer 13

Loss of characteristics asociated with original sample, progressive changes in chromosome number

Question 14

What are the two culturing cell lines ?

Answer 14

1. Suspension cells
2. Adherent cells

Question 15

What are suspension cells ?

Answer 15

1. Do not attach to culture vessel
2. Derived from immune system
   3.To subculture, dilute cells with fresh growth medium

Question 16

What are adherent cells ?

Answer 16

1. Surface proteins that adhere to culture vessels
2. To subculture use trypsin/EDTA to detach cells and dilute cells with fresh growth medium

Question 17

What is the source of serum ?

Answer 17

Foetal bovine or horse serum

Question 18

What are the disadvantages of serum ?

Answer 18

1. Not the phsiological fluid cells are in contact with in tissue
2. Protein interferance in bioassays
3. Contains serum proteases
4. Constitution not clearly defined

Question 19

What is cryopreservation ?

Answer 19

Storage of cells at -180 degrees in liquid nitrogen

Question 20

What does cryopreservation allow for ?

Answer 20

Establishment of cell repositories

Question 21

When should cells be cryopreserved ?

Answer 21

Exponential growth phase

Question 22

What should you suspend cells in in cyrotherapy ?

Answer 22

Glycerol or DMSO (10%) and 90% serum

Question 23

What is the rate of cooling of cryotherapy ?

Answer 23

1 degree C/min

Question 24

How do you prepare primary cells ?

Answer 24

1. Isolate tissue
2. Separate cell from tissue
3. Incubation and growth
4. Separate cells from other tissue components

Question 25

How can you separate cells from other tissue components ?

Answer 25

1. Explant culture 2. Mechanical 3. Enzyme dissociation

Question 26

What are the applications of cell cultures as model system ?

Answer 26

1. Cell to cell communication 2. Cellular differentiation 3. Cell cycle control 4. Phsiology or biochemistry of cells 5. Cellular response to invading pathogens

Question 27

What are the applications of cell culture ?

Answer 27

1. Model systems 2. Drug screening and development 3. Pathological studies 4. Virology 5. Biotechnology 6. Stem cell research 7. Tissue engineering 8. Gene therapy

Question 28

How can cell cultures be used in drug screening ?

Answer 28

1.Identification of potentially active compounds 2. Mechanism by which compounds exert their cytotoxic effect 3. Identification of target cell population 4. Identification of toxic concentration range 5. Relationship of drug concentration to exposure time

Question 29

What is transfection ?

Answer 29

The process of artifically introducing nucleic acids into cells

Question 30

What are the two types of tranfection ?

Answer 30

1. Transiently 2. Stable

Question 31

What are the uses of transfection ?

Answer 31

1. Expression of protein of interest 2. Bioproduction 3. Stem cell generation 4. Gene inhibition

Question 32

What are the methods used for transfection ?

Answer 32

1. Chemical gene delivery 2. Biological gene delivery 3. Physical gene delivery

Question 33

What are the reasons behind choice of transfection method ?

Answer 33

1. Dependent on cell type and experimental needs 2. High transfection efficiency 3. Low cell toxicity 4. Minimal effects on normal physiology 5. Easy to use and reproducible