Cell And Tissue Culture

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Slide 1

Cell And Tissue Culture

Mammalian Cells

Advanced Higher Biology

Slide 2

Why is it useful?

Gene manipulation

Culturing mammalian cells for cancer studies

Producing new plants through tissue culture

Slide 3

What do you need to do it?

Source of cell material

-freshly prepared

-stock of cell line

-bacterial culture

Slide 4

Suitable container

Simple flask

Sophisticated fermenter with computer-controlled monitoring

Slide 5

Growth medium

Glucose

Water

Amino acids

Salts

Slide 6

Opportunity for Gas Exchange

Oxygen

Carbon dioxide

Slide 7

Animal serum

Foetal Bovine Serum

Essential for animal cell proliferation

5% - 10% of growth media

Slide 8

Indicator

Waste products causes change in pH

Use indicator like phenol red

Changes from red to yellow

Slide 9

Control of Temperature and pH

37.5 OC

pH 7.5

Slide 10

Method for Measuring Cell Growth

Counting cell numbers in culture (haemocytometer)

Measure optical density in spectrophotometer

Slide 11

Sterilisation

Antibiotics

Sterilisation

Slide 12

Cells are either….

Anchorage – dependant

Anchorage - independant

Slide 13

Anchorage – independant cells

Cells associated with body fluid

-blood cells

Grown in suspension

Will eventually need subculturing

Slide 14

Anchorage – dependant cells

Most animal derived cells

Adhere to bottom of a flask and form a monolayer

Eventually cover entire surface of substratum (confluence)

Proliferation then stops

Need to subculture cells at this point (remove to fresh medium)