Irvine CELLine Bioreactor Flask - page 2

INTRODUCTION
Irvine Scientific specializes in developing and
manufacturing cell culture media for bioproduction.
To ensure the utility of the medium, CHO cell lines are
tested for density, viability and productivity. The cell lines
are tested at the small scale in conventional shake flasks.
Gram level production of proteins and antibodies can be
laborious using this type of
in vitro
method. Irvine Scientific
investigated a new membrane flask technology to increase
small scale production levels and to decrease culture
maintenance requirements.
ABSTRACT
Discovery scale, high-throughput methods for
in vitro
antibody and protein production can be timely and
typically requires fixed equipment such as shakers, pumps,
or stir tank reactors. This experiment evaluated membrane
flask technology that allows for high-density cell culture
as well as concentration of the expressed proteins or
antibodies. Two bioreactor flasks, the CELLine 350 and
the CELLine 1000, were evaluated and compared to shake
flask experimental controls. The CELLine 350 has a 350mL
media compartment with a 5mL cell compartment, and
the CELLine 1000 has a 1000mL media compartment
with a 15mL cell compartment. The cell compartments
are created by a gas permeable bottom membrane and an
upper dialysis membrane allowing for diffusion of media
into the cell compartment. The testing of both CELLine sizes
allowed for a comparison of scalability.
This experiment also evaluated two different media
formulations. Both the shake flask control and the CELLine
bioreactor flasks utilized either BalanCD
CHO Growth
A or IS CHO-CD G10.7 media. The shake flask cultures
were operated as both batch and fed-batch cultures (fed on
days 3-6). The CELLine bioreactor flasks had IgG harvested,
the media changed and the cells split every 7 days. Cell
concentrations, viability and IgG quantification were
measured across the 8 assessments. (2 CELLine 350s, 2
CELLine 1000s and 4 – 125mL shake flask cultures).
WHEATON
®
How Do CELLine Bioreactor Flasks Work?
CELLine bioreactor flasks utilize a dual membrane system
to create an optimal cell chamber. The chamber resides at
the bottom of the flask and is formed by a gas permeable
bottom membrane and a top dialysis membrane. The
media compartment is accessed via the green vented
cap. The cell compartment is accessed via the white cap.
The cell compartment cap gives way to a pipette septum.
This septum connects directly to the cell compartment for
simple seeding and harvest.
Media Compartment
— The media compartment allows
for bulk storage of cell culture growth medium. This
reduces the medium refreshing requirement significantly
as the media compartment is fifty times the size of the cell
compartment.
Metabolite Regulating Upper Membrane
— The
upper dialysis membrane has a 10 kDa cut off limit.
This regulates the flow of metabolites to and from the
cell compartment and retains all proteins in the cell
compartment.
Cell Compartment
— The cell compartment provides the
ideal area to inoculate and achieve high density cultures.
The compartment concentrates cells, their products, and
limits the requirement for any exogenous growth factors.
Gas Permeable Lower Membrane
— With static
cultures, gas transfer rates can be the limiting factor in
high density cultures. The CELLine bioreactor flask places
the cells directly against the gas permeable membrane to
achieve optimal levels of oxygen and carbon dioxide.
Nutrients
Glucose
Glutamine
Waste Products
Lactate
Ammonium
CO
2
O
2
10kDa Membrane
Cells
Proteins
Media
Compartment
Cell
Compartment
Media Port
Inoculation / Harvest Tube
Pipette Tip Stop
Metabolite Regulating Upper Membrane
Cell Compartment Spacer
Base
Gas Permeable Lower Membrane
Base
Silicone Membrane
Inoculation / Harvest Port
A High-Throughput Static Culture Flask
As An Alternative to Shake Flask Culture
Experiment Overview
1 3,4,5,6
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