Chapter 6: Plant and Animal Cell

Bioreactors
Function Microbial System Plant Cell System
Bulk mixing To achieve homogeneity of To achieve homogeneity of
chemical environment in biomass distribution in the
the presence of nutrient presence of sedimentation
sink and product source due to large size of cell
terms aggregates
Oxygen transfer To meet oxygen supply To meet oxygen
requirements of the requirement of the
biomass, often limiting in growing biomass without
scale up depleting the
concentration of gaseous
products below a critical
level
Shear Usually unimportant. Usually very important due
to large size of cells and
aggregates.
Mammalian Cells
 Stages in Cell Culture
• Primary cells
– Tissue taken from its original site and transferred
to an artificial medium for growth
– Requires the recurrent sacrifice of animals
• Secondary cells
– After the first passage of primary cells
– Have a finite number of passages
• Continuous or transformed cells
– Cells that can be subcultured indefinitely
 Types of Mammalian Cells
• Anchorage
Dependent cells

– Require surface
attachment to grow

– They include mostly

primary cells and
cell lines such as :

-Chinese Hamster
Ovary cells (CHO),

Baby Hamster
Kidney Cells (BHK)
and

Human Fibroblast
cells (FS-4)
• Suspension cells
– Cells not
requiring any
surface
attachment
– Examples are:
HeLa cells,
hybridoma cells
and other tumor
cells
 Some Aspects of Bioreactor Design
Design of Bio –Engineering Equipment

• Vessel Design (Height : Diameter ratio)

• Agitation
• Mechanical seal
• Stirrer
• Inlet air
• Aeration
• Ports
• Exhaust air
• Inoculation port
• Sampling valve/port
• Harvest
• Media Addition
• Air sterilization
Special Considerations for Mammalian
Cell Bioreactor
• Cells are shear sensitive thus shearing should
be minimized
• Oxygenation through bubbling may damage
cells
• Round bottom culture vessel with aspect ratio
of 2:1
 Special Modifications

Aeration and
Mixing
 Modes of Operation
Batch mode of operation

A well established production

method; generally used for vaccines The operation modes all
and biopharmaceuticals. lack in the ability to
Stirred-Tank
select or differentiate
Low yield of product
Reactor between viable and
Batch non-viable cells
Fed-batch mode

High density culture

Feed
Longer operation and higher yield
Accumulation of dead cells and debris

Continuous mode
Fed-batch Feed High density culture
Product can be harvested
Harvest continuously
Limited dilution rate
Continuous
 Perfusion Culture
 Perfusion mode
Recycle  High density culture

 Product can be harvested

Feed
continuously
Harvest  High productivity

 Dilution rate not limited

 Separation of viable and dead

cells possible
Cell separation
device

Cell Separation •Volume of the production

All Perfusion strategies
unit is smaller than
Methods in must meet the following normally required with
conditions: batch culture.
Perfusion Mode
1. Filtration
1. Stable operation •Lower investment cost on
2. Easy to scale up the bioreactor.
2. Centrifugation
3. Versatility:The process can be
3. Gravitational settling applied to any cells and any culture
medium
4. Immobilisation
 Liquid Liquid
addition withd rawal

Liquid level in
bioreacto r

Sp in filter

 Spin filter bioreactors

Perfusion Culture  Cell settler for perfusion culture
Systems  Centrifugal bioreactor
 Tangential flow or crossflow filtratio
device for perfusion culture
 Packed-bed basket perfusion culture
Fresh medium
pump
Air in

Spent medium

Celligen packed-bed
basket reactor (Wang et al.,1993)

Fibrous bed basket

Draft tube
Cell free medium
Make a comparison of an animal cell bioreactor
with a plant cell bioreactor.