Isolated population of plant single cells and method of preparing same

Abstract

This invention is a method of minimizing the variation of cell growth and production through homogeneous cell line development. To be more specific, it is the method of isolating and proliferating single cell clone from cambium to promote the stability of the plant-derived biologically active substances production by solving the problems of decrease in cell growth and the productivity during the long term culture.

1 . A method for preparing an isolated population of cells, the method comprising:
 (a) providing a tissue from a plant;   (b) obtaining from the plant tissue a tissue containing cambium;   (c) culturing in a medium the tissue containing cambium; and   (d) isolating the population of cells from the cambium,   wherein the isolated cells are innately undifferentiated.   
     
     
         2 . The method according to  claim 2 , wherein in the step (c), the medium includes auxin. 
     
     
         3 . The method according to  claim 2 , wherein the medium contains 1-3 mg/L of the auxin. 
     
     
         4 . The method according to  claim 1 , wherein the plant is the genus  Taxus.    
     
     
         5 . The method according to  claim 1 , further comprising sterilizing the plant tissue. 
     
     
         6 . The method according to  claim 1 , wherein in the step (c), the cambium is induced to proliferate cells and in the step (d), the population of cells is isolated from the cells proliferated from the cambium. 
     
     
         7 . The method according to  claim 6 , wherein in the step (c), the medium includes auxin. 
     
     
         8 . The method according to  claim 7 , wherein the medium contains 1-3 mg/L of the auxin. 
     
     
         9 . The method according to  claim 6 , wherein the plant is the genus  Taxus.    
     
     
         10 . The method according to  claim 6 , further comprising sterilizing the plant tissue. 
     
     
         11 . A method for producing a biologically active substance or substances, the method comprising the steps of:
 (a) producing the active substance or substances by culturing in a medium the isolated population of cells prepared by the method of  claim 1 ; and   (b) collecting said active substance or substances.   
     
     
         12 . The method according to  claim 11 , wherein in the step (a), a predetermined amount of the medium that has been used to culture the cells is removed and a predetermined amount of a new medium is introduced. 
     
     
         13 . The method according to  claim 11 , wherein the plant is the genus  Taxus  and the active substance is paclitaxel. 
     
     
         14 . The method according to  claim 13 , wherein the medium includes at least one selected from the group consisting of methyl jasmonate, phenylalanine, and chitosan. 
     
     
         15 . A method for preserving the isolated population of cells prepared by the method of  claim 1 , which comprises cryopreserving the isolated population of cells.