Continuous culture apparatus with mobile vessel, allowing selection of fitter cell variants and producing a culture in a continuous manner

Abstract

A method and device for growing plant, animal or stem cells in a continuous manner.

1 . A device for growing living cells in a continuous manner, comprising: 
 a flexible tubing containing culture medium and a surface in which the living cells can grow on; and    a system of clamps, each capable of open and closed positions, the clamps being positioned so as to be able to divide the tubing into: 
 i) an upstream region containing unused culture medium;  
 ii) a downstream region containing spent culture medium; and  
 iii) a growth chamber region for growing said cells disposed between the upstream and downstream regions;  
   wherein the system of clamps is constructed and arranged to open and close so as to clamp off and define the growth chamber region of the tubing between the upstream and downstream regions of the tubing, and to cyclically redefine the growth chamber region of the tubing so that a first portion of the previously defined growth chamber region becomes a portion of the downstream region of the tubing, and a portion of the previously defined upstream region of the tubing becomes a portion of the growth chamber region of the tubing.    
     
     
         2 . The device according to  claim 1 , wherein the system of clamps is structured and arranged so that each of the clamps does not move with respect to the tubing when said clamp is in the closed position.  
     
     
         3 . The device according to  claim 1 , wherein the surface is the interior surface of the tubing.  
     
     
         4 . The device according to  claim 3 , wherein the surface is a continuous support inserted inside the tubing.  
     
     
         5 . The device according to  claim 4 , wherein the means for providing a surface is a continuous fiber.  
     
     
         6 . The device according to  claim 1 , wherein the tubing is gas permeable  
     
     
         7 . The device according to  claim 1 , wherein the tubing is gas impermeable.  
     
     
         8 . The device according to  claim 1 , wherein the tubing is one of transparent and translucent to permit a turbidimeter to determine the density of the culture.  
     
     
         9 . The device according to  claim 1 , wherein the device further comprises a pressure regulator constructed to change a pressure of the growth chamber portion of the tubing relative to ambient pressure.  
     
     
         10 . The device according to  claim 1 , wherein the tubing comprises a pH indicator.  
     
     
         11 . The device according to  claim 1 , further comprising a temperature regulator constructed to allow control of a temperature of the growth chamber region of the tubing.  
     
     
         12 . The device according to  claim 1 , wherein the device further comprises an agitator constructed to allow agitation of the growth chamber portion of the tubing.  
     
     
         13 . The device according to  claim 12 , wherein the agitator comprises at least one stirring bar.  
     
     
         14 . The device according to  claim 1 , further comprising an emitter constructed to subject the growth culture chamber region to at least one of radio waves, light waves, x-rays, sound waves, an electro magnetic field, and a radioactive field.  
     
     
         15 . The device according to  claim 1 , further comprising a means for subjecting the growth chamber region to a different gravitational force.  
     
     
         16 . The device according to  claim 1 , wherein said growth chamber region comprises one or more growth chambers containing culture medium.  
     
     
         17 . A method for growing cells, comprising: 
 a) providing flexible tubing containing culture medium, and a surface in which the living cells can grow on in said tubing, and a system of clamps, each of the clamps being capable of open and closed positions, the clamps being positioned so as to be able to divide the tubing into: 
 i) an upstream region containing unused culture medium;  
 ii) a downstream region containing spent culture medium; and  
 iii) a growth chamber region for growing said cells disposed between the upstream and downstream regions; and  
   b) closing selected ones of the clamps on the tubing to define the growth chamber region of the tubing between the upstream and downstream regions of the tubing, and introducing viable cells into the growth chamber region;    c) cyclically closing and opening selected ones of the clamps to redefine the growth chamber region of the tubing so that a first portion of the previously defined growth chamber region becomes a portion of the downstream region of the tubing, and a portion of the previously defined upstream region of the tubing becomes a portion of the growth chamber region of the tubing; and    d) repeating step c) until a sufficient amount of cells have been grown.    
     
     
         18 . The method according to  claim 17 , comprising the further step of withdrawing a sample of living cells from said culture medium from the downstream region.  
     
     
         19 . The method according to  claim 17 , further comprising isolating said living cells from the downstream region.  
     
     
         20 . The method according to  claim 17 , wherein the living cells are selected from the group consisting of yeast cells, animal cells, plant cells, and stem cells.  
     
     
         21 . The method according to  claim 17 , wherein the living cells are stem cells.  
     
     
         22 . The method according to  claim 21 , wherein the living cells are selected from the group consisting of hematopoietic stem cells, bone marrow stem cells, stromal cells, astrocytes, oligidendrocytes, embryonic stem cells, fetal stem cells, umbilical cord stem cells, placenta derived stem cells, and adult stem cells.  
     
     
         23 . The method according to  claim 22 , wherein the living cells are selected from the group consisting of hematopoietic stem cells, bone marrow stem cells, stromal cells, astrocytes and oligidendrocytes.  
     
     
         24 . The method according to  claim 17 , wherein the surface in which the cells are grown is the interior surface of the tubing.  
     
     
         25 . The method according to  claim 17 , wherein the surface in which the cells are grown is a continuous fiber.  
     
     
         26 . The method according to  claim 17 , further comprising growing the cells in one or more growth chambers that are present in the growth chamber region.  
     
     
         27 . The method according to  claim 17 , further comprising growing one or more types of cells in the growth chamber region.  
     
     
         28 . The method according to  claim 17 , wherein the sufficient amount of cells of step d) is defined as a pre-determined density level of the cells.  
     
     
         29 . The method according to  claim 17 , wherein the tubing is gas permeable  
     
     
         30 . The method according to  claim 17 , wherein the tubing is gas impermeable.  
     
     
         31 . The method according to  claim 17 , wherein the tubing is one of transparent and translucent, a turbidimeter being used to determine the density level of the cells.  
     
     
         32 . The method according to  claim 17 , further comprising regulating the pressure of the growth chamber portion of the tubing relative to ambient pressure.  
     
     
         33 . The method according to  claim 17 , further comprising measuring a pH of the culture medium in the growth chamber region.  
     
     
         34 . The method according to  claim 17 , further comprising regulating the temperature of the growth chamber region with a temperature regulator constructed to control the temperature of the growth chamber region of the tubing.  
     
     
         35 . The method according to  claim 17 , further comprising agitating the culture medium in the growth chamber region with an agitator.  
     
     
         36 . The method according to  claim 35 , wherein the agitator comprises at least one stirring bar.  
     
     
         37 . The method according to  claim 17 , further comprising subjecting the growth culture chamber region to at least one of radio waves, light waves, x-rays, sound waves, an electro magnetic field, and radioactive field.  
     
     
         38 . The method according to  claim 17 , further comprising subjecting the growth chamber region to a different gravitational force.