Biology Reference
In-Depth Information
Table 1
Operational and hydrodynamic conditions for cell culture and AdV infection in stirred tank
Cell line
HEK 293
MDCK-E1
Culture type
Suspension cells
Adherent/microcarriers
Vessel characteristics
System
BioStat DCU-3
Vessel type
Univessel
®
5 L Round bottom
Univessel
®
2 L Round bottom
Total volume (L)
6.6
2.7
Working volume (L)
2-5
0.8-2
Impellers
Two 6-blade rushton turbine
Two 3-blade
Gassing system
Ring sparger
Headspace
Operational conditions
Working volume (L)
5
2
pO
2
(%)
50
a
40
b
pH
c
7.2 ± 0.05
7.4 ± 0.05
Agitation rate (rpm)
50-210
50
Aeration rate (vvm)
0.01
0.1
Temperature (°C)
37
Inoculum (10
6
cells/mL)
0.5
0.2
CCI (10
6
cells/mL)
1
1
MOI (ip/cell)
5
5
TOH (hpi)
36-72
Hydrodynamic conditions
d
Working volume (L)
5
2
Agitation rate (rpm)
50-210
50
Aeration rate (vvm)
0.01
0.1
Reynolds number (10
4
)
0.48-20
0.27
Eddy size (μm)
25-73
181
Shear stress rate (N/m
2
)
0.09-0.80
0.06
Tip speed (m/s)
0.17-0.70
0.14
a
pO
2
is controlled by varying sequentially the nitrogen partial pressure in gas inlet, agitation rate, and oxygen partial
pressure in gas inlet
b
pO
2
is controlled by varying sequentially the nitrogen and oxygen partial pressure in gas inlet
c
pH is controlled using CO
2
and 1 M NaHCO
3
d
It is known that the hydrodynamics of the culture conditions are critical for scale-up since they affect cell growth and
consequently virus production [
33
,
34
]
