WELCOME

CELL SUSPENSION
CULTURE
SUSPENSION CULTURE
:
PRINCIPLE :
 To overcome limitations of callus culture sspension
culture was initiated.

 Movement of cells in nutrient medium,

facilitates gaseous exchange,
removes any polarity of cells due to gravity,
eliminates nutrient gradients within the medium
and at the surface.
BATCH CULTURE
 Cells grow in a fixed volume of Culture medium

 Most commonly maintained in conical flask containing 25-75 ml of

liquid medium agitated on orbitary platform shakers at speed of
80-120 rpm

 Shows sigmoid growth curve over short period of time

 Ifthe culture is to be maintained a small amount of cells must be

removed and transferred to a vessel with a fresh medium
SLOWLY ROTATING CULTURE :
 Singlecells and aggregates are grown in aspecially designed flask,the
nipple flask

 Each flask have 8 nipple-like projections

 10flasks are loaded in a circular manner on a large flat disk of a

verticle shaker

 When the flask disc rotates at the speed of 1-2 rpm the cells within
each nipple of the flask is alternatively bathed in a culture medium
and exposed to air
EFFECT OF AGITATION :
 Directly
affected the growth and viability of the cells in
suspension culture due to aeration and shearing effect
At 120 rpm - Max fresh wt and max viability

At 60 rpm -cells died due to aggregation and clumping

only the cells at outer layer of the aggregate are alive

At 160 rpm - cells died due tp rupturing

SHAKE CULTURE :
 Cellsare aggregated in a fixed
volume of the liquid medium in
conlical flask

 Conicalflasks are placed on

orbitary platform shaker at
60 - 180 rpm
SPINNING CULTURE :

 Large volumes of
cells are cultured in
10L bottles,with the
bottles spinning in a
spinner at120 rpm at
an angle of 450
stirred culture :
 It is also used to culture large volumes of cells

 Large culture vessels are not rotated but the

cell suspension inside the vssels is kept
dispersed continuously by bubbling sterile air
through the culture medium

 The use of magnetic stirrer(Revolves at 200-

600 rpm) is the most convenient way to
agitate the medium
CONTINUOUS CULTURE :
 Involves continuous influx of the fresh medium
and a withdrawl of the used medium without any
cell loss
 Thus helping to increaese biomass