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P L A N T C E L L , O R G A N , A N D T IS S U E C O L L E C T IO N S , T H E M E T H O D S O F

G E R M P L A S M P R E S E R V A TIO N

CRYOPRESERVATION OF

POLYSCIAS FILICIFOLIA BAILEY.

CELLSUSPENSION CULTURE

A.S.Krivokharchenko, N.D.Chernyak, A.M.Nosov

Timiryazev Institute o fPlantPhysiology, RussianAcademy o fSciences, ul.

Botanicheskaya 35, Moscow, 127276Russia

Creation o f industrial technology o f cell culture producing biologically

active substances demands elaboration of method fo r long and safe

preservation o f initial cell lines.

The goal o f this work was to investigate the possibility of cryopreservation

o f

P. filicifo lia

cell suspension culture which is producer o f important

secondary metabolites.

The exponentially growing suspension cells o f P. filicifolia were mixed

with the cryoprotectant (sucrose and glycerol) solution and transferred into

the plastic tube o f about 0.5 ml capacity and 10-12 cm long. The tubes were

vertically placed into a chamber o f freezer equipped with a programmer and

ice crystallization was initiated 10 min later. A cooling rate was 0.4 °C/min

to -35 °C, whereupon the tubes were immersed in liquid nitrogen. The samples

were thawed in a ir at room temperature. The postthaw cell viability was 30-

35% o f initial one. The cells from the tubes were plated in the Petri dishes

(40 mm in diameter) on the surface of an agar-solidified medium. Cell growth

was observed after 10-12 days. In one month the cells were resuspended in

a liquid nutrient medium . The growth index o f the cell suspension after

cryostorage was identical to that for initial cell suspension. However, during

the first 5 subcultures the level of cell aggregation was higher in the renewed

cell suspension in comparison with the initial one.

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