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Mass Transfer considerations in Biotechnology

The mass transfer problem

The mass transfer problem

What is mass transfer?
Mass transfer is simply the movement of mass. Mass transfer can involve the
movement of mass through a fluid, movement of mass through catalyst pores or
cellular slimes or movement of mass between phases.
Before a chemical or biochemical reaction can occur, the reactants and catalysts
must meet. There must therefore be a mass transfer step before a reaction can
occur. Mass transfer is also an important consideration in downstream processing
where mass must be moved between phases.
What is the "mass transfer problem" and why is it important in biotechnology?
The problem with mass transfer in biotechnology is that mass transfer steps can be a slow
process. Since a mass transfer step must occur before a reaction can occur, then a slow rate of
mass transfer can lead to slow reaction rates. As the size of the reactor increases, mass
transfer problems likewise increase. When we design processes, we must always consider the
mass transfer rate and the factors which affect the rate of mass transfer.
There are therefore two factors which must be considered when improving a fermentation or
enzyme reaction. One factor is the catalyst (ie. the cells or enzymes). The process can be
improved by selecting or engineering a new catalyst or by changing the reactor configuration
and process control system (eg. temperature and pH). The other factor is mass transfer. The
mass transfer rate can be improved by changing the agitation system, aeration system and
reactor configuration and mode of operation.
What are some examples of process in which mass transfer rates can be slow?
Some examples include
The breakdown of oils and alkanes which require the movement of molecules
from an non-aqueous environment to the aqueous bulk liquid. Organisms and
enzymes which breakdown oil will often have hydrophobic characteristics which
allow them to directly access the oil globules and thus speed up the rate of oil
degradation.
The transfer of oxygen from bubbles to the bulk liquid. Oxygen is poorly soluble
in the fermentation medium and the rate of transfer from the gas phase to the
liquid phase can also be very slow. The trick in designing fermenters is to speed
up the oxygen transfer rate.
The degradation of viscous solutions, for example starch solutions. When a
solution is viscous, mixing becomes difficult as a result mixing in the bulk liquid
is slow reducing the probability of an enzyme-starch molecule reaction.
In a biochemical reaction system, where will mass transfer be slowest?
There are 3 areas where mass transfer rates can limit the rate of a biolgoical process
In large fermenters, the movement of molecules through the bulk liquid can be
rate limiting factor. The ideal mixing time in large reactors is normally between 28 minutes.
The movement of molecules between phases can also be a rate limiting process.
For example, the movement of oxygen from a bubble to the bulk liquid is
regarded as a rate limiting step in fermentation processes.
The movement of molecules through pores (eg. those of immobilized enzyme
systems) and the movement of molecules through microbial slimes such as exist
in immobilized cell systems.
What is meant by the statement that a reaction is "diffusion limited"?
A reaction is diffusion limited when the reaction rate is determined by the diffusion of a
substrate or reactant in a part of the reactor fluid. In this case, the reaction will be mass
transfer limited. For example, the movement of oxygen molecules through a boundary
layer occurs by diffusion. Likewise the movement of molecules through pores is also
diffusion limited.

What factors affect the rate of mass transfer?

To understand the factors that affect the mass transfer rate, we can look at the mass
transfer rate equation describing the rate of mass transfer between fluids of different
phase or through porous media such as pores or slimes.:
Mass transfer rate = dC/dt = k A (C* - C)
Measures the rate at which molecules move through pores or through
an interfacial boundary layer. Determined by
Temperature
Size of the molecules
Viscosity of the fluid
Mass transfer
k
Mixing conditions (stirrer speed and stirrer design)
coefficient
Higher temperatures increase diffusion rates. Larger
molecules diffuse slower than large molecules and the
rate of diffusion reduces with an increase in the
viscosity of liquid. Higher turbulence reduces the size
of the boundary layer.
The area is determined by the size and number of the globules/bubbles
or the size and number of immobilization particles. Small particles or
globules/bubbles provide a higher surface area to volume ratio than
large ones. Factors affecting the size of the bubbles include
the stirrer speed and type of impeller
the reactor design
the way in which the non-aqueous material is
introduced into the aqueous phase
the medium composition (eg. the presence of surface
active agents)
Area available for
A
The stirrer and the stirrer speed plays a major role in
mass transfer
breaking up globules and bubbles. A draft tube can also
be used to ensure that bubbles stay small and do not
coalesce. Another way of ensuring that
globules/bubbles remain small is to ensure that they are
small when they enter the reactor. The sparge ring for
example in aerated fermenters plays a major role in
ensuring that small air bubbles enter the reactor. The
medium can also affect the interfacial area. Oils and
silicone antifoams encourage bubbles to coalesce.
Detergents have an opposite effect.
The driving force is the gradient between the boundary layer and the
bulk liquid (average concentration) or the concentration. Factors
affecting this gradient include:
the solubility of the substrate
metabolic activity
The boundary layer in a system containing a substrate
with a higher solubility will contain a higher
C* - The diffusional
concentration of the substance. This increases the
C
driving force
diffusional forces driving substrates across the
boundary layer. In terms of the equation, this a higher
solubility increases the saturation concentration (C*).
Metabolic activity on the other hand uses the substrate
and therefore decreases the concentration in the bulk
liquid (C); again increasing the driving force across the
boundary layer.

Why are small bubbles or globules better than large ones when mass transfer
between phases is involved?
A small bubble or globule will have a larger surface area to volume ratio than a one with
a larger volume. You can try it yourself. Calculate the surface area:volume ratio of two
different spheres.
The area of a sphere is 8 x 3.14 x radius2 and the the volume is 4/3 x 3.14 x radius3
How can we speed up the mass transfer rate?
To speed up the mass transfer rate, we need to speed up each of the three components of
the mass transfer rate equation:
Mass transfer rate = dC/dt = k A (C* - C)

To increase the mass transfer coefficient, we can need to either reduce the size
of the boundary layer or increase the rate at which molecules move through the
boundary layer. This can be achieved by
Mass
k
transfer Increasing the rate of molecules Increase temperature
coefficient through the boundary layer
Decrease the viscosity of the medium
Decrease the size of the
Increase turbulence by increasing the stirrer
boundary layer
speed and improving the agitation system
The interfacial area can be increased by breaking up the globules and bubbles
or by increasing the number of globules and bubbles.
Decreasing bubble/globule size can be achieved by
Increasing the the stirrer speed and using an impeller which is
more suited for breaking up bubbles/globules (eg. a Rushton
turbine or other sort of radial flow impeller
Improving the reactor design. Baffles are used to create
Area
turbulence and shear which break up globules and bubbles. A
A available for
draft tube can be used to minimize bubble coalescence and thus
mass transfer
maintain small bubbles or globules
Breaking the bubbles/globules up before they enter the reactor.
This can be achieved by using a sparger in an aerated fermenter
or by physically and chemically breaking up globules before
they are pumped into the reactor.
With oxygen transfer, the addition of detergents can discourage
bubble/globule size coalescence. But note that this can also
cause major foam problems in aerated bioreactors.
This gradient can be increased by increasing the solubility of the substrate.
With aerated bioreactors, this can be achieved by increasing the pressure at the
The
point where the air enter (eg. by using taller reactors) and by increasing the
C* diffusional partial pressure of the oxygen in the gas (eg. by using pure oxygen instead of
C
driving force air). The solubility of oxygen can also be increased by decreasing the
temperature and by reducing the concentration of salts and sugars in the
medium (not a very practical alternative).

What do the terms in the mass transfer rate equation mean?

Mass transfer rate = dC/dt = k A (C* - C)
k is the mass transfer coefficient
A is the area available for mass transfer
(C* - C) - The driving force is the gradient between the boundary layer and
the bulk liquid (average concentration) or the concentration.
How does mixing affect the rate of mass transfer?
Mixing increases mass transfer rates by
increasing turbulence which decreases the size of boundary layers and thus speeds
up mass transfer rates.
breaking up bubbles and globules and prevents their coalescence
increasing the rate of movement of molecules through the bulk liquid

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How does temperature affect the rate of mass transfer?

Temperature affects the mass transfer rate by
changing the solubility of substrates. It may increase or decrease the solubility
and thus increase or decrease mass transfer rates
increasing the rate of molecular diffusion thus increasing the rate of movement of
molecules through boundary layers.
Mass Transfer considerations in Biotechnology
Oxygen transfer

* When considering fermentation systems, why is it that oxygen transfer is considered

to be a more important problem than the supply of other nutrients?
*

Oxygen has a much lower solubility in water than sugars and nutrients. For
example sugar dissolves up to 500 -600 g per litre while the maximum solubility
of oxygen at 1 atm/4oC in pure water is only 8 mg per litre.

Oxygen is a gas. Unlike other compounds you cannot keep add excess oxygen to
the medium. Any oxygen that is not transfered into the medium will be wasted.

Many cells are very sensitive to dissolved oxygen concentrations. A sudden drop
in the dissolved oxygen concentration can cause cells to drastically modify their
metabolism or physiology; for example, some cells may die, others may switch to
fermentation, others go into a stationary phase. Some Bacillus spp. sporulate
when dissolved oxygen concentrations fall, thus ending the vegetative cycle.

Unlike sugars and proteins, cells cannot store oxygen. Fresh oxygen must be
available for the cells at all times during the fermentation.

The transfer of oxygen is expensive. Both agitation and air compression consume
considerable amounts of energy.

* What are the major rate limiting steps in the transfer of oxygen from bubbles to
cells?
*

The movement of oxygen from through the boundary layer around a bubble

The movement of oxygen through microbial slimes (when immobilized cultures

are being used)

The movement of oxygen through the bulk liquid when mixing is poor or when
the medium is viscous.

* How is the total interfacial area between bubbles and the bulk liquid measured?
*

It is generally not measured. Instead the oxygen transfer coefficient (kl) and the
interfacial area (a) are combined into a single term referred to as kla, the oxygen
transfer coefficient per unit volume.

* What factors affect the interfacial area between the bubbles and the bulk liquid?
The interfacial area can be increased in two ways:
* increasing the number of bubbles in the reactor
* decreasing the bubble diameter
Operating factor
Effect
Radial flow impellers are more effective at breaking up bubbles
Impeller design
than radial flow impellers
Bubble break-up increases with the stirrer speed. If the impeller
Stirrer speed
speed is too low, bubbles will coalesce underneath the impeller
leading to a flooded impeller and poor oxygen transfer.

Air flow rate

Height of the reactor

Increasing the air flow rate increases the total volume of air in the
reactor
The gas hold-up increases with height of the reactor, thus increasing
the total volume of air in the reactor

Spargers should be designed such that the air is passed through

Sparger design
many small holes creating a large number of small bubbles in the
reactor
Antifoams accumulate around the gas-liquid interface encouraging
Antifoams (and detergents)
bubbles to coalesce, thus reducing the interfacial area

* What factors affect the oxygen transfer coefficient?

The oxygen transfer coefficient is determined by the rate at which oxygen molecules pass
through the boundary layer. This can be increased by reducing the size of the boundary
layer and by increasing the rate of movement of molecules through the boundary layer.
Effect
Operating variable
This is achieved by increasing the level of
turbulence by increasing stirrer speed and
Decreasing the size of the bubble boundary layer impeller size. Increasing turbulence also
increases the rate of movement of
dissolved oxygen through the bulk liquid.
Increasing temperature causes molecules to
move faster, thus the rate of diffusion
Increasing the rate of movement of oxygen through through the boundary layer increases.
the bubble boundary layer
Excess antifoam will accumulate
around the gas-liquid interface and
hinder the movement of oxygen.
* What factors affect the oxygen concentration gradient across the bubble boundary
layer?
The dissolved oxygen concentration gradient across the bubble boundary layer (Co* Co)is the driving force which pushes the oxygen out of the bubble. This concentration
gradient can be increased by increasing the saturation concentration of oxygen (Co*) by
increasing the partial pressure of oxygen in gas phase. This can be achieved by:
* using pure oxygen rather than air
* using taller reactors and thus increasing the total pressure at the base of the
reactor.
Factors which will decrease the saturation concentration of oxygen include:
* Increased concentrations of sugars and salts
* Higher temperatures
Note that cellular activity will decrease Co and thus also increase the oxygen transfer rate.
* How does agitation affect the oxygen transfer rate?
Agitation plays four major roles in improving the oxygen transfer rate.
* The agitation system is used to break apart bubbles and thus increase the
interfacial transfer area (a).
* Agitation moves the bubbles sideways and thus increases the gas holdup.
* In large reactors or when the medium is viscous, agitation plays an important role
moving dissolved oxygen molecules through the bulk liquid.
* Increasing the agitation rate reduces the size of the boundary layer surrounding
bubbles and thus increases the oxygen transfer coefficient (k)
* Why are "small bubbles" regarded as the secret to a successful fermentation?
Small bubbles have a higher surface to volume ratio than large bubbles. This increases
the interfacial area for oxygen transfer and thus increases kla and the oxygen transfer rate.

* What does the expression "bubble coalescence" mean?

Bubble coalescence occurs when bubbles meet. When two bubbles meet, they can form a
single bubble. The presence of antifoams in the medium will increase the tendency of
bubbles to coalesce. Ionic detergents on the other hand will reduce bubble coalescence.
* Why are antifoams added during a fermentation and what are the problems
associated with antifoams?
Antifoams are added to prevent the formation of foam. Foam accumulates during aerobic
fermentations. The problem with antifoams is that they also reduce the kla of the oxygen
transfer system and thus reduce the oxygen transfer rate.
* Why are antifoams and detergents called surface active agents?
Antifoams and detergents are called surface active agents because they act at the surface
of the liquid. Water molecules are bound together by hydrogen bonding forces. At the
same time, water and air do not attract each other and can be thought of as repelling each
other. (Water is hydrophilic and air is hydrophobic).
* Why do foams form in fermentation systems and what problems do they cause?
Foams form in fermenters because cells release detergent like substances. During a
fermentation, cells will proteins and other detergent like compounds into the medium.
These compounds can include extracellular enzymes and metal binding compounds. As
cells die, they release their DNA and intracellular proteins. One indication that a
fermentation has ended is the excessive accumulation of foam or an increase in the
antifoam requirement. Foams are a problem in aerated fermenters. When the headspace
becomes full of foam, the pressure in the reactor will build up. This pressure will try to
push the foam out of the fermenter. As a result. air filters will be damaged and in some
cases, the covers and ports which are not screwed down can actually be blown off the
fermenter. This can cause injury to personnel and can also cause a loss of the fermenter
contents and contamination of the surroundings.
* How can foam formation be controlled?
Foam can be controlled in three ways
These include
* Reducing the air flow rate
Preventative
* Reducing the addition of detergents (eg. Tween 80, commonly
measures
used to harvest fungal spores can be replaced with glycerol)
Antifoams and oils are often added to fermentations. Antifoams are based on
silicone oil formulations. Vegetable oils such as peanut oil, whilst not as
Antifoams
effective as silicone antifoams are natural products and cause less problems in
downstream processing.
* Physical foam breakers can be used. These include:
* the application of heat at the surface. This causes the
bubbles to expand and thus break.
* mechanical breakers which physically break the foam.
* vibratory systems likwise cause the foam to break.
Physical methods
* Increasing the size of the disengagement zone in bubble column
and air-lift and fluidized bed bioreactors.
* A draft tube will reduce foam production, turbulence at the
surface and thereby assisting in the breakdown of the foam. An
air-lift reactor with an internal riser is said to produce less foam
than one with an external riser.

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Bioreactor design

Basic bioreactor design considerations

How do cells in "standing cultures" receive oxygen?
Oxygen transfer in standing cultures occurs through the surface of the culture.
Why do we incubate T-flasks in a horizontal position?
By laying T-flasks in a horizontal position, we increase the surface area available for
oxygen transfer.
What is surface air entrainment?
When the surface of a medium becomes agitated, small "waves" will form trapping small
bubbles of air. This phenomenom is known as air entrainment and will plays an important
role in oxygen transfer in some types of shake flask mixers and non-sparged stirred tank
reactors.
What are the problems associated with using shake flasks for performing laboratory
experiments?
Shake flasks suffer from the following problems:
pH control is either very inefficient or not possible and as most microorganisms
do change the pH of their medium, the results obtained from shake flask
experiments are always questionable.
Oxygen transfer is very poor compared to aerated stirred tank bioreactors and thus
microbial cells will often be oxygen limited during the fermentation
Shake flask conditions do not simulate actual operating conditions
It is difficult to sample from shake flasks while the flasks are shaking. If shaking
is stopped for sampling, the lack of oxygen can lead to cell death or a change in
cell metabolism.
What are the advantages of air-lift reactors over bubble-column reactors?
The main difference between air-lift and bubble-column reactors is that the former
contain a draft tube. The draft tube gives the airlift reactor a number of advantages:
It prevents bubble coalescence by causing bubbles to move in one direction
It equally distributes shear stresses throughout the reactor and thus provides a
healthier environment for cell growth.
The enhanced flow cyclical movement of fluid through the reactor increases heat
transfer rates from the heat transfer jacket.
Why do gas-mixed reactors have a larger height to diameter ratio than stirred tank
bioreactors?
Gas mixed reactors do not have impellers to decrease bubble diameter and increase kLa.
To compensate for this, gas mixed reactors are designed with a larger height to diameter
ratio so as to improve oxygen transfer efficiency by:
increasing the pressure at the base of the reactor and thus increase the saturation
concentration of oxygen at the base (Co*).
increasing the time the bubbles spend in the reactor and thus increase the bubble
residence time and gas hold-up. This increases the air volume in the reactor
available for oxygen transfer and increases the time available for each bubble to
transfer to the medium.

What limits the height of a bioreactor?

As the bubbles rise, they transfer out oxygen and receive carbon dioxide from the
medium. If the reactor is too tall then, bubbles at the top will be rich in carbondioxide and poor in oxygen. The cells at the top of the reactor will therefore be
starved of oxygen.
Mixing time also increases with the reactor height. Thus a reactor which is too tall
will from mass transfer problems with cells in some areas being starved of oxygen
for relatively long periods of time. A large mixing time also makes process
control difficult. For example, the pH may vary throughout the reactor, making it
difficult to determine when to add alkali and how much alkali to add.

Construction costs also increases as the reactor height increases. For stirred tank
reactors, the cost of stabilizing a very long shaft limits the economic feasibility of
using very tall reactors.

Why do airlift bioreactors often produce higher productivities than gassed stirred
tank bioreactors?
The major reason for the higher productivities of airlift bioreactors is that the draft tube
distributes shear forces throughout the reactor. As a result, cells are not exposed to large
variations in shear forces and thus are able to grow in a more stable physical environment. In
contrast, in stirred tank reactors, high shear conditions will arise near the impeller causing
cell damage or cell stress and thus lowering productivity.

The stirred tank bioreactor

Why should stirred tank bioreactors and tanks in general be fitted with a sump?
Tanks are normally fitted with a sump to all easy drainage of solids. These solids could
be microbial cells or fine precipitates which during the emptying of the tank, begin to
settle out. The sump acts as a collection point towards which these solids will move and
thus can be readily removed. This assists in the cleaning of the tank.
Why do we use the expression "vvm" when expressing air flow in fermenters?
vvm is the calculated as the air flow rate divided by the volume of the fermenter. It is
used as a rudimentary way of scaling up fermenter operation; particularly in the case of
air-lift reactors. For example, if pilot scale experiments demonstrate that an air flow rate
of 2 vvm gives optimal results, an air flow rate of 2 vvm will also be used in the larger
commercial scale reactor.

Aeration and Agitation in Animal Cell Cultures

What is "shear"
Shear has two components. Shear stress which is the force per unit area acting on a body and
shear rate which a measure of how the velocity changes as we move away from that body. Shear
can be visuallized as liquid flow lines moving at different speeds and directions. Such velocity
fluctuations will occur in turbulent eddies. The smaller the eddy and the greater the velocity
fluctuation then the greater will be the level of shear.
Are animal cells shear sensitive?
Yes and no. Unlike bacteria, animal cells do not possess a cell wall which should protect them
from shear forces. However, their small size will protect them from shear forces that arise in the
bioreactor liquid. Their small size in effect allows them to ride between the the turbulent eddies.
Despite this bioreactors and cell culture systems must be designed to minimize shear problems.
These design considerations should include:
* The use of low shear axial flow impellers
* The use of shear protectorants such as serum or Pluronic F-68
* Ensuring that the medium contains sufficient nutrients to ensure that cells are not
nutritionally stressed.
* Selecting cells that are not shear sensitive
* Minimizing impeller speeds
Cells can be damaged by shear forces in foams which may accumulate on the surface of the
medium. Cells trapped between moving bubbles can be pulled apart by the moving bubbles.
How does the nutritional state affect the shear sensitivity of animal cells
Studies performed during the 1980's showed that many of the media then used for cultivation of
mammalian cells contained insufficient nutrients, in particular amino acids. It was found that by
increasing the concentrations of necessary amino acids, the susceptibility of cells to shear stress
was reduced dramatically.

How can we determine if animal cells have been physically damaged?

When cells have been physically damaged, they will release their cytoplasmic and membrane
enzymes into the medium. The level of cell damage can therefore be determined by measuring
the concentration of some of these enzymes. An enzyme which is often analysed for is L-lactate
dehydrogenase. This is found on the cell membranes and the high LDH activity will indicate a
high
level of cell damage. The enzyme is also stable and its activity is easy to measure.
Why are filamentous fungal cells more shear sensitive than bacterial cells?
Both bacteria and fungi are protected by cell walls. Bacteria are typically more able to survive
and grow in the high shear environment of a sparged stirred tank reactor because:
* The cell walls of filamentous fungi is composed of chitin, which is not as strong as the
peptidoglycan bag which protects bacterial cells.
* Filamentous fungi are also larger than bacterial cells and therefore are more susceptible to
shear forces. The Komologorov eddy size required to cause cell damage is much higher
than that required for bacterial cells.
How do changing shear forces affect cells?
Shear forces in a stirred tank bioreactor. Near a typical radial flow impeller, shear forces will be
considerably higher than in other parts of the reactor. Cells growing in a stirred tank bioreactor
will therefore be subjected to changing shear forces as they move within the reactor. These
changing shear forces have a negative effect on cell growth. Cells have been found to produce
higher biomass yields in the more uniform shear environment of an airlift bioreactor as compared
to stirred tank bioreactor environment.
How do bubbles damage animal cells?
Bubbles damage amimal cells in two main ways. When bubbles collapse at the surface of a
reactor, cells trapped in the wake of the bubble will be subjected relatively high stress forces.
These forces are strong enough to rupture the cell's membrane. The shear forces can originate
from the velocity gradient arising from the upward movement of the bubble and the downward
jet resulting from the bursting bubble.
One way of protecting cells from bubble damage is to add surface active agents which stabilize
the bubbles. This gives cells time to detach from the bubble before it bursts. Some surface active
agents are believed to make bubbles "slippery" such that cells do not attach to them. Pluronic
F68 and serum are typically used as shear protectorants.
Long high molecular weight substances such as dextran which increase the viscosity of the
medium will also protect the cells against cell damage. The high viscosities absorb the kinetic
energies of the liquids.
Cells can also be damaged when trapped in foams. As the foams move, they will tend to drag the
membranes of trapped cells with them. When bubbles surrounding a cell move in different
directions or at different speeds, the cell will be torn or sheared apart. Bubble damage can be
reduced ensuring that foam layers are kept small. This can be achieved by increasing the
diameter of the disengagement zone, by using slow aeration rates or by removing the foam
problem altogether with bubble-free bioreactors.
Which do more damage, big bubbles or small bubbles
Large bubbles have a larger surface area kept in place by a larger total surface energy than small
bubbles. When the bubbles collapse, this surface energy together with the bubbles kinetic and
pressure energy would be released. Accordingly collapsing large bubbles should cause more cell
damage than smaller bubbles.
However, when the phenomenom of bubble damage was first discovered, conflicting data as to
whether big bubbles did in fact cause more damage than small bubbles. The reason for this was
that the phenomenom of cell damage in foams was not understood. Small bubbles have a larger
surface area to volume ratio and would be able to trap more cells than large bubbles. Small
bubbles can thus do more foam damage than large bubbles.
Why are animal cell bioreactors normally constructed to maximum volumes of only 1000
litres?
Animal cell culture is an expensive task. The man-power, media and materials required to make
each litre of cell culture medium will cost far more than the cost preparing microbial cell media.
The contamination of large animal cell bioreactor can therefore represent a significant loss for a

company. Companies often prefer to limit the size of their bioreactors and thus limit the size of
the risk.
What is Pluronic F-68 and how does it protect cells from damage
Pluronic F-68 is a non-ionic detergent which acts as a "shear protectorant". It is composed
primarily of polypropylene glycol (capped with polyethylene glycols). It protects cells from
shear damage and from bubble damage. It works by acting on the surface properties of the cell
culture medium and possibly the cell surface.
It is believed to
* make the bubbles slippery by providing a highly mobile bubble boundary layer such that
the cells do not attach to bubbles
* strengthen the cells membrane
* stabilizes foam. This allows the cells to detach from the bubbles before they burst and
thus protecting them from the forces released when the bubbles collapse.