Chapter 6: Plant and Animal Cell Bioreactors

Function Bulk mixing

Microbial System To achieve homogeneity of chemical environment in the presence of nutrient sink and product source terms To meet oxygen supply requirements of the biomass, often limiting in scale up

Plant Cell System To achieve homogeneity of biomass distribution in the presence of sedimentation due to large size of cell aggregates To meet oxygen requirement of the growing biomass without depleting the concentration of gaseous products below a critical level Usually very important due to large size of cells and aggregates.

Oxygen transfer

Shear

Usually unimportant.

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 lack in the ability to and biopharmaceuticals.
Stirred-Tank Reactor

Low

yield of product

Batch

Fed-batch mode
High

select or differentiate between viable and non-viable cells

density culture operation and higher yield of dead cells and debris
Continuous mode

Feed

Longer

Accumulation

Fed-batch

Feed

High

density culture

Product

Harvest

can be harvested continuously

Limited

dilution rate

Continuous

Perfusion Culture

Recycle Feed Harvest

Cell separation device

Perfusion mode High density culture Product can be harvested continuously High productivity Dilution rate not limited Separation of viable and dead cells possible
Volume of the production unit is smaller than normally required with batch culture. Lower investment cost on the bioreactor.

Cell Separation Methods in Perfusion Mode

1. Filtration

All Perfusion strategies must meet the following conditions:

1. Stable operation 2. Easy to scale up 3. Versatility:The process can be applied to any cells and any culture medium

2. Centrifugation
3. Gravitational settling 4. Immobilisation

Liquid addition

Liquid withd rawal

Liquid level in bioreacto r Sp in filter

Perfusion Culture Systems

Spin filter bioreactors Cell settler for perfusion culture 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

Cell free medium

Draft tube

Make a comparison of an animal cell bioreactor with a plant cell bioreactor.