Broth Agar 3.38 x 106 4.2 x 106 3.30 x 106 6.5 x 106 a
Potato Dextrose Agar 1.93 x 106 2.1 x 106 3.56 x 106 1.31 x 107 a
Sodium Azide
Tryptic Soy Broth Agar
between 50 -100 x 106 cells,
depending on the cell type.
Note: Volumes below are for 50 -100 x 106 cells, harvested from one confluent 500 cm2 dish.
Remove medium from cells. Wash 50 -100 x
Thus, if 200 x 106 hybridoma cells were inoculated,
about 48-72 hours later 400 x 106 will reside in the cell compartment. To assess the number of cells, pipette the volume
up and down 3 or 4 times to
96%.
Cell Density
(cells/mL)
Cell Viability
(%)
Flask 1
Flask 2
Flask 3
2.32 x 106 92.1
3.12 x 106
2.76 x 106 86.3
91.8
To demonstrate the kinetic growth curve of expresSF+® cells
in
g. Re-suspend the cells at a concentration of
3 x 106 to 5 x 106 cells/ml in 50% fresh Gene
Therapy Medium-3 and 50% conditioned Gene
Therapy Medium-3. Supplement the medium with
DMSO at a final concentration
production of IL-2 ������� pg
IL-2/106 Jurkat cells. The PHA control culture yielded
106 Jurkat cells. The control with no
addition was 50 pg IL-2/106 Jurkat cells.
References
1. Weiss
minutes at
200 x g. Re-suspend the cells at a
concentration of 3 x 106 to 5 x 106 cells/ml in
50% fresh Gene Therapy Medium-1 and 50%
conditioned Gene Therapy Medium-1.
Supplement the medium with
°C (70°F), 50% relative humidity, out of direct sunlight, in original packaging
Ordering:
Part Number Format Presentation
IST-106-078LR** LabRoll™ 1 Roll 610M x 78 mm
IST-106-078SR** Sterile
Split cells when density is 3–4 × 106 cells/ml.
Note: Overdilution will result in cell death; avoid densities less than 0.2 × 106 cells/ml.
Trypan Blue Exclusion Method
1. Add 100 µl cells to 100
minutes at 200 x g.
Re-suspend the cells at a concentration of 3 x 106 to
5 x 106 cells/ml in 50% fresh Gene Therapy Medium-
2 and 50% conditioned Gene Therapy Medium-2.
Supplement the medium with DMSO
the
viable cell density reaches 1.5×106 cells/mL
• Before proceeding to the next step (reduced serum concentration), a minimal viable cell
density of 1.5×106 cells/mL and a viability > 95% should
showed the
best cell growth in the GTM-3 with 6.5 x 106 cells/ml at day seven of the culture.
Figure 3. Comparison of virus production from Per.C6® cells in GTM-3 and GTM-1. Per.C6® cells were seeded
0.5-1 x 106 viable cells/mL when
they reach 3-6 x 106 viable cells/mL in fresh EX-Cell®
CD Insect Cell Medium to a volume of 20-50 mL in a
125 mL shake flask.
5. Incubate the flask for 3-4
pfu/cell) 1 x 10-4 , 5 x 10-4, 1 x 10-3 5 x 10-4
Time of infection
(days after cell inoculation)
2, 3, 4 4 (or cell density > 2 x 106 vc/mL)
In Mobius® 3
cultured for batch growth in
Mobius 3 L and 50 L Single-Use Bioreactors. While cells were seeded in the
50 L bioreactor at 1 106 viable cells/mL (vc/mL) and in the 3 L bioreactor at
5 105 vc/mL
6 × 105 vc/mL and diluted
with fresh media on Day 3. Peak viable cell density (VCD) of 8 × 106 vc/mL was obtained with
viability >90% in Mobius 50 L bioreactor. Comparable growth of cells was achieved
of 5 x 105 to 1 x 106 cells/ml. Next, harvest
and re-seed the cells at 1.5 x 105 cells/ml in the basal
medium containing 5% FBS. At subsequent passage,
split the cells into a 50:50 ratio of the basal