390
Related Techniques
mosquito cells, collagen-based micro spheres (Verax) were used. The inoculum level was 1 x 10 6 cells/ml beads. The spinner culture (Fig. 3) was
performed at 28°C in a CO 2 incubator where the CO 2 concentration was
adjusted to 5%. Using this set up, a maximum cell number of 2 x 10 7
cells /ml beads was obtained. Because the ratio of beads to medium (v/v)
was 40%, and the micro spheres were not fully loaded with cells, much
higher cell densities per volume of bioreactor may be possible.
I Subprotocol 2
Shaking Flask Culture
For some anchorage-independent cell lines, a shaking flask culture may
be used.
Materials
Equipment - Reciprocal shaker
- Erlenmeyer shaker flasks (500 ml, and 1 and 21)
Solutions - Culture medium
Procedure
1. Harvest cells from a stock culture.
2. Count the cells.
3. Dispense the cells into the shaker flasks so as to make a cell density of
3 x 10 5 cells/ml.
4. Set the shaker flasks on a reciprocal shaker.
5. Operate the shaker at 140 r.p.m.
Results
Nguyen et al. (1993) cultured Sf-9 cells in serum-free Ex-cell 400 medium or IPL-41 medium supplemented with lipids and yeastolate at 27°C
using this type of culture system. The cell density increased from 3 x 10 5
to 3 x 10 6 cells after 3 days propagation in either medium.
I Subprotocol 3
Spinner Culture
This type of culture system is used for relatively small-scale cultures.
Materials
Equipment - Spinner flasks (Fig. 3)
Related Techniques
mosquito cells, collagen-based micro spheres (Verax) were used. The inoculum level was 1 x 10 6 cells/ml beads. The spinner culture (Fig. 3) was
performed at 28°C in a CO 2 incubator where the CO 2 concentration was
adjusted to 5%. Using this set up, a maximum cell number of 2 x 10 7
cells /ml beads was obtained. Because the ratio of beads to medium (v/v)
was 40%, and the micro spheres were not fully loaded with cells, much
higher cell densities per volume of bioreactor may be possible.
I Subprotocol 2
Shaking Flask Culture
For some anchorage-independent cell lines, a shaking flask culture may
be used.
Materials
Equipment - Reciprocal shaker
- Erlenmeyer shaker flasks (500 ml, and 1 and 21)
Solutions - Culture medium
Procedure
1. Harvest cells from a stock culture.
2. Count the cells.
3. Dispense the cells into the shaker flasks so as to make a cell density of
3 x 10 5 cells/ml.
4. Set the shaker flasks on a reciprocal shaker.
5. Operate the shaker at 140 r.p.m.
Results
Nguyen et al. (1993) cultured Sf-9 cells in serum-free Ex-cell 400 medium or IPL-41 medium supplemented with lipids and yeastolate at 27°C
using this type of culture system. The cell density increased from 3 x 10 5
to 3 x 10 6 cells after 3 days propagation in either medium.
I Subprotocol 3
Spinner Culture
This type of culture system is used for relatively small-scale cultures.
Materials
Equipment - Spinner flasks (Fig. 3)