Animal cell Bioreactors

Interest in the in vitro cultivation of animal cells has developed

because of the need for large scale production of monoclonal
antibodies, hormones, vaccines and other products
 What is Bioreactor?
• An apparatus for growing organisms (yeast, bacteria, or
animal cells) under controlled conditions.
• Used in industrial processes to produce
pharmaceuticals, vaccines, or antibodies
• Also used to convert raw materials into useful by
products such as in the bioconversion of corn into
ethanol.
• Bioreactors supply a homogeneous (same throughout)
environment by constantly stirring the contents.
• Bioreactors give the cells a controlled environment by
ensuring the same temperature, pH, and oxygen levels.
Required properties of bioreactors
• Simplicity of design
• Large number of organisms per unit volume
• Uniform distribution of micro-organisms
• Simple and effective oxygen supply
• Low energy requirement
• Uniform distribution of energy
• providing information about the formation of
3D tissue
Organisms growing in bioreactors may
be:
• Suspended/Anchorage independent
• Immobilized/Anchorage dependent .
A simple method, where cells are
immobilized, is
Conditions imposed by the bioprocess
Petri dish with agar gel.
Large scale immobilized cell bioreactors
are:
• moving media, also known as
Moving Bed Biofilm Reactor (MBBR);
• Packed bed;
• Fibrous bed;
• Membrane.
 Anchorage Dependent cells
• Anchorage dependent cells: These cells require a solid support
for their replication. They produce cellular protrusions
(pseudopodia) which allow–Require surface attachment to
grow them to adhere to positively charged surfaces and often
grow as monolayer.

• Anchorage independent cells: These cells do not require a

support and can grow as a suspension in submerged culture.
Established and transformed cell lines are normally in this
category
 Modified Stirred fermenter
• The modifications made to fermenters are to
reduce the possibility of cell damage due to
shear, heat or contamination.
• Marine propellers revolving at a slow speed
(10 to 100 rpm) will normally provide
adequate mixing.
• Hemispherical bottoms on the vessels will
ensure better mixing of the broth at slow
stirrer speeds.
• Water jacket heating is often preferred since
heating probes may give rise to localized Range of culture apparatus for
zones of high temperature which might suspension cells (A) magnetic bar
damage some of the cells. spinner culture, (B) Techne MCS
• Magnetic driven stirrers may be used to stirrer, (C) surface stirrer , (D) small
reduce the risk of contamination. scale fermenter with marine
impeller, (E) airlift fermenter, (F)
vibro-fermenter .
 Air lift fermenter
Air-lift fermenters have
proved ideal for growth of
some cell lines because of the
gentle mixing action and
reduced shear forces when
compared with those in
stirred vessels. The absence
of a stirrer and associated
seals excludes a potential
source of contamination
Example :monoclonal Range of culture apparatus for suspension
antibodies production from cells (A) magnetic bar spinner culture, (B)
hybridoma cells Techne MCS stirrer, (C) surface stirrer , (D)
small scale fermenter with marine
impeller, (E) airlift fermenter, (F) vibro-
fermenter .
 Microcarriers
• Microcarriers may provide a solution
to growing anchorage-dependent
cultures by providing the necessary
surface for attachment.
• Animal cells normally have negative
surface charge and will attach to a
charged surface by electrostatic forces.
• DEAE Sephadex A-50 resin beads and
number of microcarrier beads,
manufactured from dextran, cellulose,
gelatin, plastic or glass, are
commercially available and used.
 Encapsulation
• Encapsel, a technique developed by Damon Biotechnology, U.S.A.,traps the animal
cells in sodium alginate spheres which are then coated with polylysine to form a
semi-permeable membrane. The enclosed alginate gel is solubilized with sodium
citrate to release the cells into free suspension within the semipermeable coat.
• The cells are entrapped in calcium alginate or agarose beads which will allow high
molecular weight products to diffuse into the medium
 Hollow fibre
chambers
• Anchorage dependent cells
cultured in the bundles of
hollow fibres held together in
cartridge chambers.
• The cells are grown in the extra
capillary spaces (ECS) within the
cartridge.
• Medium and gases diffuse
through from the capillary lumea
to the ECS.
• Monoclonal antibodies, viruses,
gonadotropin, insulin and
antigens
 Perfusion Culture

• High density culture

• Product can be harvested

continuously

• High productivity

• Dilution rate not limited

• Separation of viable and dead

cells possible
 Packed glass bead reactors

•Packed glass bead reactors have

proved to be useful for long term
culture of attached dependent cell
lines.
• Increasing the size of vessels causes
problems with mass transfer of
oxygen and nutrients and scale up can
be achieved by increasing the number
of small vessels.