Suspension Culture

Suspension Culture


• Suspension Culture

• Definition

• Types

• Apparatus used


At the end of the lecture the student will able to:

• Discuss the technique of suspension culture

• Discuss the various suspension culture apparatus

Suspension Culture

Suspension Culture

• Type of culture - single cell or small aggregates of cells multiply while suspended in agitated liquid medium under aseptic conditions

• Cell suspension - friable callus cells – liquid medium – agiated – rotary shaker

• Agitation of medium is very important, it helps in two ways

i. Prevent the clumping or settling of cells – dispersion – auxins or pectinase or cellulase enzyme

ii. Gaseous exchange, aeration

• Mainly of two types

Batch suspension culture and Continuous suspension culture

Batch suspension culture and Continuous suspension culture

Batch suspension culture:

• Cell material or inoculums grow – finite volume of agitated liquid medium

Continuous suspension culture:

• Process in which spent medium is continuously replaced by fresh medium in order to maintain steady growth

• Depletion of nutrients does not occur due to continuous flow of fresh medium

Suspension culture apparatus:

i) Agitated liquid culture apparatus

a. Shake culture                   

b. Slowly rotating culture (Auxophytons) - Tumble tube Nipple flask Spin culture

ii) Stationary liquid culture apparatus – Chemostat and Turbidostat

Shake Culture:

• Agitation is provided by moving the liquid medium

• Consists a metal platform and number of clamps

• Clamps are used to hold container

• When platform shakes, it provides circular motion to medium and leads to gaseous exchange or aeration

• Shaker speed – 30 – 150 rpm

Advantages: Large amount of cells can be grown, various sized flasks can be used

Disadvantage: Sometimes evaporation of medium due to high speed

Tumble tube:

• Consists of glass tube of capacity 10 ml with a neck at the centre

• Neck is plugged with cotton (gaseous exchange)

• 24 such tubes are placed on a circular disc, 5-6 circular discs are placed on a shaft

• When shaft rotates, tubes tumble, medium flows from one end to another

• As the tube tumbles, explant alternatively exposed to medium and air, which helps in gaseous exchange

• Large amount of cells cannot be grown

Nipple flask:

• Modified form of tumble tube

• Available in 2 sizes, 250 ml with 8 nipples and 1000 ml with 10 nipples

• About 10 nipple flasks are placed on a circular disc

• When disc rotates, medium enters and leaves the projection, explant alternatively exposed to medium and air Speed of rotation – 1-2 rpm

• Large amount of cells can be grown

Nipple flask:

Spin culture:

• Consists of bottle of capacity 10 Liter

• Mounted at an angle 450 in the spin

• Bottle spins because of the inclined angle

• As a result there is a movement of medium

• Movement leads to gaseous exchange

• Large amount of cells can be grown compare to tumble tube and nipple flask


• It is a continuous suspension culture

• Consists air inlet, air outlet, provision for continuous supply of medium and removal of spent medium, and magnetic stirrer

• There is a fixed rate of media input and it is time programmed

• A steady state is achieved by the addition of the medium



• It is also a continuous suspension culture

• Consists similar apparatus as that of chemostat along with densitometer

• Turbidity of suspension culture medium increases with increase in number of cells

• Turbidity is measured by measuring optical density using densitometer

• Once turbidity reaches maximum level spent medium is removed and fresh medium enters

Growth measurement:

• Cell count/cell number

• Cell viability

Packed cell volume:

• Suspension culture medium with cells is subjected to Centrifugation at 2000 rpm for 10-15 min

• Cells settled down at the bottom         

• Volume of these settled cells is measured – Packed cell volume           


• Tissue culture is growing of tissues or cells on a suitable nutrient medium under aseptic conditions in vitro, may be plant or animal tissue culture

• Types include callus, suspension, hairy root, single cell, haploid, organogenesis, embryo culture etc

• Suspension culture is growing of cells/cell aggregates in liquid media, batch or continuous

• Agitation is essential, helps in gaseous exchange

• Initiated using friable callus cells, sterile seedling, embryo etc

• Agitated liquid apparatus like shake culture, tumble tube, nipple flask, spin culture and stationary liquid culture like chemostat and turbidostat

• Large amount of cells can be grown compare to callus culture

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