Cell culture

Question 1

What is cell culture?

Answer 1

• Lab technique used to grow and maintain cells in vitro
• cells are removed from an animal or plant and grown in an artificial environment
• cells are cultured in an artificial culture medium using aseptic confition

Question 2

Aseptic meaning

Answer 2

• sterile/sterilised conditions to ensure free from contamination

Question 3

What is an in vitro cell culture?

Answer 3

allows individual cells to grow when provided with sufficient nutrients and growth factors

Question 4

What are homogenous cultures?

Answer 4

-genetically identicall cultures
derived from a single parental cell (clone)
- cells can divide in culture until limited by a parameter e.g a lack of nutrients

Question 5

What are heterogeneous cultures?

Answer 5

• genetically varied cultures

Question 6

What is cell confluence?

Answer 6

• the percentage area covered by cells in a culture dish or flask
• cells need to be subcultured (passaged) by transferring them to a new dish with fresh growth medium to provide more room for continued growth
• once subcultured- known as cell line

Question 7

What are primary cell cultures?

Answer 7

• cells freshly isolated from organ tissue and maintained for growth in vitro
• Cells proliferate under appropriate conditions until they reach confluence

Question 8

What are the 2 basic systems for growing cells in culture?

Answer 8

1.) As monolayers on an artificial substrate (adherent culture)
2. Free-floating - in the culture medium (suspension culture)

Question 9

What is required for cell culture conditions?

Answer 9

• essential nutrients (Amino acids, carbs, vitameins, minerals)
• Growth factors
• Hormones
• Gases (O2, CO2)
• Regulated physico-chemical environment
• pH
• osmotic pressure
• temperature

Question 10

What are some applications of cell culture?

Answer 10

• major tool used in cellular and molecular biology
• model systems for studying the normal physiology and biochemistry of cells
• effects of drugs and toxic compounds on the cells
• used in drug screening and development
• large scale manufacturing of biological compounds
  -(vaccines, therapeutic proteins)

Question 11

What is the general layout of cell cultures?

Answer 11

• clean and uncluttered
• in fume cupboard- further towards the centre of the hood the better
• vertical flow of filtered air

Question 12

What are some advantages of cell culture?

Answer 12

• consistency and reproducibility of results using cell culture
• homogenous population in contrast to using animal tissue
• heteroenous population of cells (different stages of growth/viability?
• toxicology- less expensive in cell culture rather than using whole animals (also ethical issues)

Question 13

What are disadvantages of cell cultures?

Answer 13

• During growth, the characteristics of some cells in cultur can change
• Intracellular changes may result (enzyme activities e.g.)
• Cells in culture can’t always mimic in vivo tissue microenvironment - results using cell culture assays
• have to be confirmed using in vivo assays