Method for controlling proliferation of cord blood hematopoietic stem cells and use thereof

Abstract

The present invention provides a method for controlling the proliferation and differentiation of cord blood-derived hematopoietic stem cells with excellent safety when proliferating them by culturing. The hematopoietic stem cells are inoculated into a medium containing a sonicated liquid component of cord blood. The proliferation and differentiation of the cord blood hematopoietic stem cells can be inhibited in the presence of the sonicated liquid component of cord blood. On the contrary, the proliferation of cord blood hematopoietic stem cells can be accelerated by inoculating the hematopoietic stem cells into a medium containing a non-sonicated liquid component of cord blood. Thus, according to the present invention, by using serum derived from cord blood, it is possible to regulate the inhibition of the proliferation and differentiation of cord blood hematopoietic stem cells and the acceleration of the proliferation of the same as desired.

1 . A method for controlling proliferation of cord blood hematopoietic stem cells, the method comprising the step of:
 (X) inhibiting proliferation and differentiation of hematopoietic stem cells by culturing the hematopoietic stem cells in a medium containing a sonicated liquid component of cord blood.   
     
     
         2 . The control method according to  claim 1 , further comprising the step of:
 (Y) accelerating proliferation of the hematopoietic stem cells by culturing the hematopoietic stem cells in a medium containing a non-sonicated liquid component of cord blood.   
     
     
         3 . The control method according to  claim 2 , wherein,
 in the step (X), proliferation and differentiation of the hematopoietic stem cells are inhibited, and then,   in the step (Y), the proliferation of the hematopoietic stem cells inhibited in the step (X) is accelerated.   
     
     
         4 . The control method according to  claim 2 , wherein,
 in the step (Y), proliferation of the hematopoietic stem cells is accelerated, and then,   in the step (X), proliferation and differentiation of the hematopoietic stem cells proliferated in the step (Y) are inhibited.   
     
     
         5 . The control method according to  claim 1 , wherein the hematopoietic stem cells and the liquid component are derived from cord blood of the same individual. 
     
     
         6 . The control method according to  claim 5 , wherein the individual is a mammal. 
     
     
         7 . The control method according to  claim 6 , wherein the mammal is a human. 
     
     
         8 . A method for producing cord blood hematopoietic stem cells, the method comprising the step of:
 controlling proliferation of hematopoietic stem cells by the control method according to  claim 1 .   
     
     
         9 . The production method according to  claim 8 , wherein proliferation and differentiation of the hematopoietic stem cells are inhibited by culturing the hematopoietic stem cells in the medium containing the sonicated liquid component of cord blood until a desired time at which proliferation of the hematopoietic stem cells is started. 
     
     
         10 . The production method according to  claim 8 , wherein the hematopoietic stem cells are proliferated, and then, proliferation and differentiation of the proliferated hematopoietic stem cells are inhibited by culturing the proliferated hematopoietic stem cells in the medium containing the sonicated liquid component of cord blood until a desired time at which the hematopoietic stem cells are used. 
     
     
         11 . The production method according to  claim 8 , wherein proliferation of the hematopoietic stem cells is accelerated by culturing the hematopoietic stem cells in a medium containing a non-sonicated liquid component of cord blood. 
     
     
         12 - 19 . (canceled) 
     
     
         20 . A cord blood component preparation device for preparing cord blood hematopoietic stem cells, a sonicated liquid component of cord blood, and a non-sonicated liquid component of cord blood from the same cord blood, for use in the method for controlling proliferation of cord blood hematopoietic stem cells according to  claim 1 , the cord blood component preparation device comprising:
 a storage unit for storing cord blood;   an erythrocyte accommodation unit for accommodating erythrocytes;   a hematopoietic stem cell accommodation unit for accommodating hematopoietic stem cells;   first and second plasma accommodation units for accommodating plasma; and   first and second serum accommodation units for accommodating serum,
 wherein the storage unit is connected to each of the erythrocyte accommodation unit, the hematopoietic stem cell accommodation unit, and the first and second plasma accommodation units, 
 the first plasma accommodation unit is connected to the first serum accommodation unit, 
 the second plasma accommodation unit is connected to the second serum accommodation unit, 
 the storage unit is configured so that an erythrocyte sedimenting agent can be introduced thereto, 
 the erythrocyte accommodation unit is configured so that erythrocytes having sedimented from cord blood in the storage unit can be introduced thereto, 
 the hematopoietic stem cell accommodation unit is configured so that hematopoietic stem cells having sedimented from a supernatant obtained after the sedimentation of the erythrocytes in the storage unit can be introduced thereto, 
 the first plasma accommodation unit and the second plasma accommodation unit are configured so that the supernatant obtained after the sedimentation of the erythrocytes and the supernatant obtained after the sedimentation of the hematopoietic stem cells in the storage unit can be introduced thereto, respectively, 
 the first plasma accommodation unit is configured so that sonication and a fibrin-forming treatment with respect to the supernatant introduced thereto can be performed therein, 
 the first serum accommodation unit is configured so that, among a sediment fraction and a supernatant fraction obtained from the supernatant after being subjected to the sonication and the fibrin-forming treatment in the first plasma accommodation unit, the supernatant fraction can be introduced thereto as sonicated serum, 
 the second plasma accommodation unit is configured so that a fibrin-forming treatment with respect to the supernatant introduced thereto can be performed therein, and 
 the second serum accommodation unit is configured so that, among a sediment fraction and a supernatant fraction obtained from the supernatant after being subjected to the fibrin-forming treatment in the second plasma accommodation unit, the supernatant fraction can be introduced thereto as non-sonicated serum. 
   
     
     
         21 . A cord blood component preparation device for preparing cord blood hematopoietic stem cells, a sonicated liquid component of cord blood, and a non-sonicated liquid component of cord blood from the same cord blood, for use in the method for controlling proliferation of cord blood hematopoietic stem cells according to  claim 1 , the cord blood component preparation device comprising:
 a storage unit for storing cord blood;   a cell-containing plasma accommodation unit for accommodating plasma containing hematopoietic stem cells;   a hematopoietic stem cell accommodation unit for accommodating hematopoietic stem cells;   first and second plasma accommodation units for accommodating plasma; and   first and second serum accommodation units for accommodating serum,   wherein the storage unit is connected to the cell-containing plasma accommodation unit,   the cell-containing plasma accommodation unit is connected to each of the hematopoietic stem cell accommodation unit and the first and second plasma accommodation units,   the first plasma accommodation unit is connected to the first serum accommodation unit,   the second plasma accommodation unit is connected to the second serum accommodation unit,   the storage unit is configured so that an erythrocyte sedimenting agent can be introduced thereto,   the cell-containing plasma accommodation unit is configured so that, among a sediment fraction containing erythrocytes having sedimented from cord blood in the storage unit and a supernatant fraction, the supernatant fraction can be introduced thereto as plasma containing hematopoietic stem cells,   the hematopoietic stem cell accommodation unit is configured so that hematopoietic stem cells having sedimented from the plasma containing hematopoietic stem cells in the cell-containing plasma accommodation unit can be introduced thereto,   the first plasma accommodation unit and the second plasma accommodation unit are configured so that a supernatant obtained after removing the hematopoietic stem cells in the cell-containing plasma accommodation unit can be introduced thereto,   the first plasma accommodation unit is configured so that sonication and a fibrin-forming treatment with respect to the supernatant introduced thereto can be performed therein,   the first serum accommodation unit is configured so that, among a sediment fraction and a supernatant fraction obtained from the supernatant after being subjected to the sonication and the fibrin-forming treatment in the first plasma accommodation unit, the supernatant fraction can be introduced thereto as sonicated serum,   the second plasma accommodation unit is configured so that a fibrin-forming treatment with respect to the supernatant introduced thereto can be performed therein, and   the second serum accommodation unit is configured so that, among a sediment fraction and a supernatant fraction obtained from the supernatant after being subjected to the fibrin-forming treatment in the second plasma accommodation unit, the supernatant fraction can be introduced thereto as non-sonicated serum.   
     
     
         22 . A cord blood component preparation device for preparing cord blood hematopoietic stem cells, a sonicated liquid component of cord blood, and a non-sonicated liquid component of cord blood from the same cord blood, for use in the method for controlling proliferation of cord blood hematopoietic stem cells according to  claim 1 , the cord blood component preparation device comprising:
 a storage unit for storing cord blood;   a cell-containing plasma accommodation unit for accommodating plasma containing hematopoietic stem cells;   first and second plasma accommodation units for accommodating plasma; and   first and second serum accommodation units for accommodating serum,   wherein the storage unit is connected to the cell-containing plasma accommodation unit,   the cell-containing plasma accommodation unit is connected to each of the first and second plasma accommodation units,   the first plasma accommodation unit is connected to the first serum accommodation unit,   the second plasma accommodation unit is connected to the second serum accommodation unit,   the storage unit is configured so that an erythrocyte sedimenting agent can be introduced thereto,   the cell-containing plasma accommodation unit is configured so that, among a sediment fraction containing erythrocytes having sedimented from cord blood in the storage unit and a supernatant fraction, the supernatant fraction can be introduced thereto as a plasma fraction containing hematopoietic stem cells, and after the plasma fraction introduced thereto is separated into a sediment fraction containing the hematopoietic stem cells and a supernatant fraction, the cell-containing plasma accommodation unit can be divided into a lower region containing the sediment fraction and an upper region containing the supernatant fraction,   the lower region of the cell-containing plasma accommodation unit serves as a hematopoietic stem cell accommodation unit,   the first plasma accommodation unit and the second plasma accommodation unit are configured so that the supernatant fraction in the upper region of the cell-containing plasma accommodation unit can be introduced thereto,   the first plasma accommodation unit is configured so that sonication and a fibrin-forming treatment with respect to the supernatant introduced thereto can be performed therein,   the first serum accommodation unit is configured so that, among a sediment fraction and a supernatant fraction obtained from the supernatant after being subjected to the sonication and the fibrin-forming treatment in the first plasma accommodation unit, the supernatant fraction can be introduced thereto as a sonicated serum fraction,   the second plasma accommodation unit is configured so that a fibrin-forming treatment with respect to the supernatant introduced thereto can be performed therein, and   the second serum accommodation unit is configured so that, among a sediment fraction and a supernatant fraction obtained from the supernatant after being subjected to the fibrin-forming treatment in the second plasma accommodation unit, the supernatant fraction can be introduced thereto as non-sonicated serum.   
     
     
         23 . A cord blood component preparation device for preparing cord blood hematopoietic stem cells, a sonicated liquid component of cord blood, and a non-sonicated liquid component of cord blood from the same cord blood, for use in the method for controlling proliferation of cord blood hematopoietic stem cells according to  claim 1 , the cord blood component preparation device comprising:
 first and second storage units for storing cord blood;   an erythrocyte accommodation unit for accommodating erythrocytes;   a hematopoietic stem cell accommodation unit for accommodating hematopoietic stem cells;   first and second plasma accommodation units for accommodating plasma; and   first, second, and third serum accommodation units for accommodating serum,   wherein the first storage unit is connected to each of the erythrocyte accommodation unit, the hematopoietic stem cell accommodation unit, and the first and second plasma accommodation units,   the first plasma accommodation unit is connected to the first serum accommodation unit,   the second plasma accommodation unit is connected to the second serum accommodation unit,   the first storage unit is configured so that an anticoagulant and an erythrocyte sedimenting agent can be introduced thereto,   the erythrocyte accommodation unit is configured so that erythrocytes having sedimented from cord blood in the storage unit can be introduced thereto,   the hematopoietic stem cell accommodation unit is configured so that hematopoietic stem cells having sedimented from a supernatant obtained after the sedimentation of the erythrocytes in the storage unit can be introduced thereto,   the first plasma accommodation unit and the second plasma accommodation unit are configured so that the supernatant obtained after the sedimentation of the erythrocytes and the supernatant obtained after the sedimentation of the hematopoietic stem cells in the storage unit can be introduced thereto, respectively,   the first plasma accommodation unit is configured so that sonication and a fibrin-forming treatment with respect to the supernatant introduced thereto can be performed therein,   the first serum accommodation unit is configured so that, among a sediment fraction and a supernatant fraction obtained from the supernatant after being subjected to the sonication and the fibrin-forming treatment in the first plasma accommodation unit, the supernatant fraction can be introduced thereto as sonicated serum,   the second plasma accommodation unit is configured so that a fibrin-forming treatment with respect to the supernatant introduced thereto can be performed therein,   the second serum accommodation unit is configured so that, among a sediment fraction and a supernatant fraction obtained from the supernatant after being subjected to the fibrin-forming treatment in the second plasma accommodation unit, the supernatant fraction can be introduced thereto as non-sonicated serum,   the second storage unit is configured so that a blood coagulation-accelerating substance for accelerating blood coagulation can be introduced thereto, and   the third serum accommodation unit is configured so that a supernatant fraction obtained after blood coagulation in the storage unit can be introduced thereto.