2. Scaleup Producing more cells and/or more volume
of a product, usually across several stages
Suspension
Monolayer
Primarily involves an increase in the volume
Primarily involves an increase in the surface
area
Stirred suspension
Static suspension
3. Factors
Appropriate Scaleup- the physical and chemical
requirements of cells have to be satisfied
Physical :
1. Configuration of the bioreactor
2. Supply of Power
3. Stirring of the medium
Chemical parameters
1.Medium and Nutrients
2. Oxygen
3. pH and buffer systems
4. Removal of waste products
4. Scale up
1 × 109
– 1 × 1010
cells -- produced in simple
stirrer cultures of around 1-10-L capacity
larger-scale cultures of 1 × 1011
– 1 × 1012
cells will require
apparatus ranging from a 100-L laboratory scale fermentor
to a semi-industrial pilot plant with capacities of from 100 to
1000 L
Monolayer- increased growth surface area
Large surface area flasks
245mm square dishes
Roller bottles
Multilayer culture chambers
Hyper Flask vessels
E-cube bioreactors
5. Scale Up- Suspensions
Increase inVolume
Agitation of the medium - depth exceeds 5
mm,
Above 5–10 cm, sparging with CO2 and air is
required to maintain adequate gas exchange.
Stirring of such cultures is best done slowly with a
magnet encased in a glass pendulum or with a large-
surface area paddle
7. Continuous flow culture
It is possible to keep the cells at a desired set
concentration, and maintain
The flow rate of the medium addition can be determined from
the growth rate of the culture
More susceptible to contamination
8. Air–lift bioreactor
The same air is used to supply
oxygen, provide agitation and
remove heat
It is the design to couple heat and
mass transfer process using a single
vehicle, viz., low pressure air
9. Suitable for fragile animal as well as plant cells.
Extensively used in the biotechnology industry
The bubbles formed move up to agitate and aerate the culture.
They carry the liquid along with them and release at the top
Airlift bioreactors: (a) draft-tube internal-loop configuration, (b) a split-
cylinder device, and (c) an external-loop system.
10. Scale-up in Monolayer cultures
Anchorage
dependent
Necessary to increase the surface area of
the substrate
Advantages
1. Change of the medium and washing of cells
easy
2. Easy to perfuse immobilized monolayer cells
3. Cell product formation is much higher, same set
up and apparatus can be repeatedly used with
different media and cells
Disadvantages
Tedious and costly; require more space, growth can’t
be monitored effectively, difficult to measure control
parameters
11. Roller Bottles
Goerge Gey (1933)- John Hopkins University
490cm2
to 1750cm2
Advantages:
Medium is gently and constantly agitated; surface area is
high for cell growth
12. Cell growth can still be determined using inverted
microscopes
Very economical for cultivating large quantities of anchorage
dependent cells
Prevents gradients from forming within the
medium; Provides superior gas exchange
Collection of the supernatant medium is easy
Disadvantages
Require more incubator space than flasks or dishes
Require a moderate amount of labor to produce large no. of
cells
May cause cell attachment problems
13. Cell stack chambers
Multilayered vessels provide
both a space and labor saving
Easy scaleup- by adding more layers or more chambers, requires
les space, can be stacked, no additional equipment required
Larger chambers are heavy when filled, more
difficult to manipulate, and require strong shelves
Disadvantages
Hard to check cell growth by microscopy
17. Perfused Monolayer
culture
Growth surface areas are perfused to facilitate medium
replacement and improved product formation and recovery
Fixed bed
reactors
Bed of glass beads; medium is perfused upwards
through the bed. Cells are grown on the surface of
the beads
Porous ceramic matrix with micro-channels also used
Fluidized bed reactors:
The beads are suspended in a stream of medium,
beads are porous in nature, made up of ceramics or
ceramics mixed with natural products such as
collagen
beads are low-density float in the medium. The flow
rate is equal to the sedimentation rate of the beads.
Cells can grow as monolayers on the outer surfaces and inside
18. Perfusion Bioreactor
- Is a system that cultivates cells in a controlled environment
by continuously supplying fresh medium and removing spent
medium
- Allows continuous nutrient supply and waste removal
20. E- cube Small perfused bioreactor system
It contains oxygenator,
medium reservoir, multiple
access ports- all required
tubing and fittings and uses
a disposable 10 stack cell
cube module
21. Scale-up in Microcarrier
culture
Monolayers can be grown on small spherical carriers or micro-
beads (80-300pm diameter)
Made from – Plastic
Glass
Gelatin
Collagen
Cellulose
DEAE Dextran
Micro-beads provide maximum surface area (depends on the
size and density of the beads).
It can be treated as suspension culture
22. Factors affecting microbead carrier culture
Composition and coating of beads (gelatin and
collagen beads are preferred)
Higher stirring speed is usually required
Glass beads are used when microcarriers need to be recycled
Requires less labour than dishes, flasks, roller
bottles, higher yields due to continuous perfusion of
cells with medium, easily scaledup , requires less
space than flasks and roller bottles
No direct microscopic viewing of cells; requires more
expertise to use than flasks, requires an initial
investment, requires a peristaltic pump
Limitations
