AEM - Invitrogen

Viable Cells (x10 5)
Adenovirus Expression Medium (AEM)
Growth Medium for PER.C6 ® Cells
GIBCO ® Adenovirus Expression Medium
(AEM) is a new medium optimized
for cell growth and production of adeno
and influenza viruses in PER.C6 ® cells.
Serum-free, phenol red-free, and free
of animal-origin components, it is
formulated specifically for the adaptation,
growth, and expansion of PER.C6 ®
human embryonic retinoblast cells.
Typical results for suspension culture
shaker flask conditions are shown in
figure 1.
Typical Growth Curve for
PER.C6 ® Cells in GIBCO ® AEM
35
30
25
20
15
10
5
0
0 1 2 3 4 5
Days
Figure 1. Total cell counts were conducted electronically
on the Coulter Z2 Particle
Count and Site Analyzer. Viabilities were
determined on a hemocytometer via the
trypan blue exclusion method. The data
represented above is the mean ± SD of
15 observations. The cells were seeded at
3 ✕ 10 5 cells/ml in a shaker flask.
• Recommended for PER.C6® cells
• Increases virus production and yield
• Each lot tested for cell growth
• Easy adaptation to suspension growth
• Animal-origin-free
A producer cell for the manufacture
of gene therapy vectors and vaccines,
the PER.C6 ® cell line should be
cultured in a medium that is scalable
and meets downstream purification
and regulatory requirements. AEM
is used by PER.C6 ® cell developer,
Crucell Holland B.V. for this purpose.
Together, the two technologies—cells
and medium—provide researchers
with the opportunity to produce
molecules in accordance with many
regulatory guidelines.
GIBCO ® AEM is a suitable replacement
for GIBCO ® 293 Medium, previously the
medium of choice for this
application. Able to support the
growth of PER.C6 ® cells in a variety
of culture platforms including shaker
flasks, roller bottles, spinner flasks, and
bioreactors (figures 2 and 3),
it is shown to produce adenovirus
(figure 4) and influenza viruses.
This innovative medium allows for easy
transition from adherent to suspension
culture. It also supports HEK 293 cell
growth and protein production, and
is useful for vaccine production and
gene therapy.
P R O D U C T N E W S
Recommended for Use
with PER.C6 ® Cells
GIBCO ® Adenovirus Expression
Medium has been accepted and used
by PER.C6 ® cell developer Crucell
and other end-users. AEM supports
peak cell densities over 4 ✕ 10 6/
ml following four days in culture
(figure 3). Maximal peak cell density
is twice that of 293 SFM II. Each
lot of AEM is performance-tested by
Invitrogen for cell growth.
With cell growth and virus production,
AEM is demonstrated to be the overall
best medium for PER.C6 ® cells.
Invitrogen is an official licensee of the
PER.C6 ® cell for media development.
GIBCO ® Adenovirus Expression Medium
represents an enabling technology that
helps users to maximize experimental
results with less labor-intensive
suspension cultures while eliminating
many of the concerns associated
with the use of serum and other
animal-origin materials. It is available in
1 L bottles and in versions customized
to specific user needs.
Technical Information
For additional technical information
about Adenovirus Expression
Medium, please visit our website at
www.invitrogen.com/bioproduction or
contact Cell Culture technical Services,
1-800-955-6288.

Cell Density (viable cells/ml)
4.76 10 6
4.48 10 6
4.20 10 6
3.92 10 6
3.64 10 6
3.36 10 6
3.08 10 6
2.80 10 6
2.52 10 6
2.24 10 6
1.96 10 6
1.68 10 6
1.40 10 6
1.12 10 6
8.40 10 5
5.60 10 5
2.80 10 5
Ordering Information
Description Catalog No. Size
Adenovirus Expression Medium (AEM) 12582-011 1,000 ml
Growth medium for PER.C6® cells.
Related Products
Growth Analysis
250-ml shake flasks (n=12)
490-cm 2 roller bottles (n=11)
250-ml spinner flasks (n=4)
PDT: 33, 34, 75 hrs respectively
0 1 2 3 4 5 6 7 8 9
Days in Culture
Figure 2. Comparison of cell growth of
PER.C6® cell growth in roller bottles, shaker,
and spinner flasks. PER.C6® cells were seeded
at 3.5 ✕ 10 5 viable cells/ml. Samples were
taken on the days indicated to monitor density
and viability. Each data point represents the
average viable cell number. PER.C6® cells
grew to maximum viable cell density in roller
bottles of 3.92 ✕ 10 6 at day 6 of the growth
assay. Population Doubling Times (PDT)
were determined between day 2 and day 4
for all conditions.
TrypLE Select 12563-011 100 ml
An animal-origin-free cell dissociation enzyme. 12563-029 500 ml
L-Glutamine-200 mM (100X), liquid 25030-149 20 ml
Supplied at 29.2 mg/ml in 0.85% NaCl. 25030-081 100 ml
GlutaMAX -I Supplement 35050-061 100 ml
Contains the dipeptide L-Alanyl-L-Glutamine.
www.invitrogen.com
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Tel: 1 760 603 7200 • Tel (Toll Free): 1 800 955 6288 • Toll Free Fax: 1 800 331 2286 • E-mail: tech_service@invitrogen.com
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Tel: +44(0) 141 814 6100 • Fax: +44(0) 141 814 6260 • E-mail: eurotech@invitrogen.com
Cell Density (Viable cells/ml)
4.2 10 6
3.9 10 6
3.6 10 6
3.4 10 6
3.1 10 6
2.8 10 6
2.5 10 6
2.2 10 6
2.0 10 6
1.7 10 6
1.4 10 6
1.1 10 6
8.4 10 5
5.6 10 5
2.8 10 5
Bioreactor Growth
These products are for research use, and where appropriate, as raw material components in further cell culture manufacturing applications. They are not intended for human or
Printed on recycled paper. animal diagnostic, therapeutic, or other clinical uses, unless otherwise stated. PER.C6 is a trademark of Crucell Holland B.V.
©2005 Invitrogen Corporation O-064137-r2 US 1005
AEM
293 SFM II
PDT: 28 and 26 hrs respectively
0 1 2 3 4 5 6 7 8
Days in Culture
Figure 3. Comparison of PER.C6® cell growth
in AEM and 293 SFM II under bioreactor
conditions. Cells were seeded at 3 ✕ 10 5 cells/
ml in either a 3.8 L or 5 L bioreactor.
The cells were passaged at least three times
before being examined. Population doubling
times (PDT) were determined between days
2 and 4.
VP/seeded cell
Virus Yields of PER.C6 ® Cells
in GIBCO ® AEM on Day 4 Post-
Infection
7 10 4
6 10 4
5 10 4
4 10 4
3 10 4
2 10 4
1 10 4
0
A B C
Lot
D E
Figure 4. Yields of Ad5 from PER.C6® cells
in AEM, calculated from the HPLC AEX
results. Cultures were seeded at 5 ✕ 10 5 cells/
ml, and infected at an MOI of 60.
The averages of duplicate samples taken
on day 4 are presented for each medium lot.
(Data courtesy of Crucell Holland B.V.)