Biology Reference
In-Depth Information
Fig.
4
Seed train strategies for: (
a
) HadV and (
b
) CAV-2 production
from cells grown in Erlenmeyer flasks (
see
Subheading
3.1.1
) or
from a previously inoculated 1 L bioreactor following
Subheading
3.3.1
and adapted for 1 L working volume (
see
Fig.
4
).
At this point, the bioreactor should have already 80 % of the
culture medium (4 L) with the selected culture conditions stabi-
lized (
see
Subheading
3.3.1
).
1. Harvest cells from pre-inoculation vessels and determine cell
concentration (
see
Subheading
3.2.1
).
2. Centrifuge the corresponding volume of cells to ensure an
inoculum of 5 × 10
5
cells/mL at 300 ×
g
for 10 min at room
temperature (
see
Note 33
).
3. Suspend cells in culture medium in order to bring the bioreactor
volume to final working volume (20 % of working volume).
4. Confirm cell concentration and transfer cells to inoculation flask.
5. Connect the inoculation flask to the bioreactor and let them
enter by gravity (
see
Note 34
).
