Chapter 36
Growth Rate
The growth rate of cell cultures is expressed by growth curves or population doubling time.
Subprotocol 1
Growth Curves
Data to draw growth curves can be obtained by several methods.
Replicate Tissue Culture Method
Materials
PROTOCOL
- Culture vessels (culture tube, culture bottle, culture dish, or mulch- Equipment
well plate)
- Pasteur pipettes
- Automatic dispenser
- Hemocytometer
- Culture medium
Solutions
- Physiological solution
Procedure
1. Harvest cells from the culture to be examined by suspending the cells Seeding
in the culture medium, and centrifuging the suspension at 150 g for 5
min.
2. Wash cells with physiological solution by centrifugation at 150 g for
5 min.
3. Make a homogeneous cell suspension by suspending cells in culture
medium. The density of cells should be adjusted to between 10 4 and
10 5 cells/ml.
4. Make a replicate culture by dispensing a definite volume (proper volume for vessel used) of the cell suspension into culture vessels. The
number of cultures to be made is three times the counting frequency
plus approximately 10 % of the frequency in case of accidental contamination.