Cell Disruption
Prof. Dr. Layla Othman Farhan
Cell Disruption
• Cell disruption is the process of obtaining intracellular fluid via
methods that open the cell wall.
• The overall goal in cell disruption is to obtain the intracellular
fluid without disrupting any of its components
• The method used may vary depending on the type of cell and its
cell wall composition. Irrespective of the method used, the main
aim is that the disruption must be effective and the method
should not be too harsh so that the product recovered remains
in its active form.
Types of cell disruption methods
• There are two types of cell disruption method which are
following
• Mechanical methods
• Non Mechanical methods
Mechanical Methods
• Mechanical methods are those methods which required
some sort of force to separate out intracellular
protein without adding chemical or enzyme
1. Mortar & pastel/grinding
2. Blender
3. Bead beating
4. Ultra sonication
5. Homogenization
Mortar & Pestle
• Just give the cells a good old
grinding.
• This does not have to be in
suspension and is often done with
plant samples frozen in liquid
nitrogen.
• When the material has been
disrupted, metabolites can be
extracted by adding solvents.
Blenders
• The use of blenders (high
speed can be used to
disrupt cell walls.
• This is the same process
used by centrifugation,
which separates or
concentrates materials
suspended in a liquid
medium
Bead beating
• Glass or ceramic beads are
used to crack open cells
• but this kind of mechanical
shear is gentle enough to keep
organelles intact.
• It can be used with all kinds of
cells, just add beads to an
equal amount of cell
suspension and vortex
Ultra sonication
• Ultrasonic homogenizers work by inducing vibration in a
titanium probe that is immersed in the cell solution.
• A process called cavitation occurs, in which tiny bubbles
are formed and explode, producing a local shockwave and
disrupting cell walls by pressure change.
• This method is very popular for plant and fungal cells but
comes at a disadvantage: It’s very loud and has to be
performed in an extra room
Ultra sonication
Homogenization
• Liquid-based homogenization is the most widely used cell
disruption technique for small volumes and cultured cells.
• Cells are lysed by forcing the cell or tissue suspension
through a narrow space
• Homogenizers use shearing forces on the cell similar to
the bead method. Homogenization can be performed by
squeezing cells through a tube that is slightly smaller than
beads beating.
Non Mechanical Methods
• Non mechanical methods are further divided into three class which
are following
• Physical methods
1. Freeze thaw
2. Microwave/ Thermolysis
3. Osmatic shock
4. Electric discharges
• Chemical methods
• Enzymatic methods
Freeze Thaw
This method used when working with soft plant material
and algae.
freezing is used to achieve cell disruption via a series of
freezing and thawing cycles.
Freezing forms ice crystals, which expand upon thawing,
and this ultimately causes the cell wall to rupture.
Microwave
• Microwave (along with autoclave and other high
temperature methods) are used to disrupt the bonds
within cell walls, and also to denature proteins. This is a
somewhat risky method, as the excess heat can quickly
damage the rest of the cell.
Osmatic shock
• Through the process of osmosis, water can be moved into
the cell causing its volume to increase to the point that it
bursts.
• Note that this method can only work with animal cells
and protozoa, since they do not have cell walls.
Electrical Discharges
• It is also possible to achieve cell disruption via electrical
discharges in mammalian cells.
• Cells that are bounded by plasma membranes and, unlike
plant cells, have no cell wall.
• This method allows researchers to examine secretion by
exocytosis, which is a process during which the
membrane-bounded sphere (intracellular vesicle) shifts to
and fuses with the plasma membrane
Chemical Method
• Often used with plant cells (and sometimes in
combination with shearing), organic solvents
such as toluene, ether, benzene, methanol,
surfactants, and phenyl ethyl alcohol DMSO can
be used to permeate cell walls.
• EDTA can be used specifically to disrupt the cell
walls of gram negative bacteria, whose cell walls
contain lipopolysaccharides that are stabilized by
cations like Mg2+ and Ca2+. EDTA will chelate the
cations leaving holes in the cell walls.
Enzymatic methods
• Enzymes such as beta(1-6) and beta(1-3) glycanases,
proteases and mannase can be used to disrupt the cell
wall.
• This method is particularly useful for isolating the cell
without the wall (protoplast).
