Cell culture method, cell culture device, method for counting subsject matters to be counted in container and device for counting

Abstract

Cell aggregate formation control and cell aggregate disintegration control are carried out with respect to cells in a culture container to increase cell proliferation efficiency. The number of the cells in the culture container is counted without disassembly of a culture system and irrespective of the density of the cells. An external force is applied to the culture container to carry out at least one of cell aggregate formation control and cell aggregate disintegration control with respect to cells in the culture container, thus culturing the cells. The external force is applied to the culture container by pressing an agitating member onto a top surface of the culture container placed in flat orientation to a predetermined pressing degree, and moving the agitating member in a horizontal direction at a predetermined speed, thus controlling at least one of cell aggregate formation and cell aggregate disintegration with respect to the cells in the culture container. A method for counting counted targets disposed in a liquid enclosed in a container includes adjusting a thickness of at least a part of the container. At least a part of the adjusted part is set as a measurement target region. A number of counted targets in the measurement target region is counted.

1 . A method for culturing cells using a culture container, the method comprising applying an external force to the culture container to carry out at least one of cell aggregate formation control and cell aggregate disintegration control with respect to cells disposed in a culture solution in the culture container. 
     
     
         2 . The method for culturing cells according to  claim 1 , wherein the applying step of the external force to the culture container comprises:
 pressing an agitating member onto a top surface of the culture container placed in flat orientation to a predetermined pressing degree; and   moving the agitating member in a horizontal direction at a predetermined speed, whereby the culture solution in the culture container is agitated by the applying step of the external force to the culture container, thus controlling at least one of cell aggregate formation and cell aggregate disintegration with respect to the cells in the culture container.   
     
     
         3 . The method for culturing cells according to  claim 1 , wherein the cell aggregate formation is carried out with respect to cells seeded in the culture container at a start of culturing. 
     
     
         4 . The method for culturing cells according to  claim 1 , wherein the cell aggregate disintegration controls an aggregate of the cells to a size within a predetermined range. 
     
     
         5 . A cell culturing apparatus comprising:
 a loading base on which a cell container is placed, the cell container enclosing a culture solution comprising a culture medium and cells; and   an agitating member configured to press the culture container to a predetermined pressing degree and movable at a predetermined speed in a horizontal direction,   wherein the agitating member is configured to move to agitate the culture solution in the culture container so as to control at least one of cell aggregate formation and cell aggregate disintegration with respect to the cells in the culture container.   
     
     
         6 . The cell culturing apparatus according to  claim 5 , wherein the agitating member has a roller shape. 
     
     
         7 . The cell culturing apparatus according to  claim 5 , further comprising:
 a photographing device configured to photograph the cells in the culture container;   determination means for receiving an image photographed by the photographing device and for determining whether an aggregate of the cells has a size within a predetermined range; and   a driving device configured to, following a determination of the determination means, move the agitating member in the horizontal direction at the predetermined speed with the predetermined pressing degree so as to carry out the cell aggregate formation when the size of the aggregate of the cells is below the predetermined range, and configured to move the agitating member in the horizontal direction at the predetermined speed with the predetermined pressing degree so as to carry out the cell aggregate disintegration when the size of the aggregate of the cells is above the predetermined range.   
     
     
         8 . The cell culturing apparatus according to  claim 5 , further comprising a partition member configured to divide the culture container into two or more compartments comprising a culture portion and an expansion portion, and configured to expand a capacity of the culture portion in accordance with an increase in a number of the cells in the culture portion, the agitating member being configured to agitate the culture medium in the culture portion. 
     
     
         9 . A method for counting counted targets disposed in a liquid enclosed in a container, the method comprising:
 adjusting a thickness of at least a part of the container; setting at least a part of the adjusted part as a measurement target region; and   counting a number of counted targets in the measurement target region.   
     
     
         10 . The method for counting counted targets disposed in a container according to  claim 9 , further comprising, prior to adjusting the thickness of at least a part of the container, agitating the liquid in the container to equalize the counted targets in the liquid. 
     
     
         11 . The method for counting counted targets disposed in a container according to  claim 9 , further comprising, based on the counted number of the counted targets, calculating a density of the counted targets in the liquid, and/or calculating a number of counted targets throughout the liquid. 
     
     
         12 . The method for counting counted targets disposed in a container according to  claim 9 , the counting step of the number of the counted targets comprises counting the number of the counted targets in a photographed image of the measurement target region. 
     
     
         13 . The method for counting counted targets disposed in a container according to  claim 9 , further comprising reducing a thickness of at least a part of the container when the number of the counted targets in the measurement target region is equal to or more than a predetermine value, and then counting the number of counted targets. 
     
     
         14 . The method for counted targets in a container according to  claim 13 , wherein the reducing step of the thickness of at least a part of the container comprises pressing or stretching the container using a thickness adjusting member. 
     
     
         15 . The method for counting counted targets disposed in a container according to  claim 9 , further comprising increasing a thickness of at least a part of the container when the number of the counted targets in the measurement target region is less than a predetermine value, and then counting the number of counted targets. 
     
     
         16 . The method for counting counted targets disposed in a container according to  claim 15 , wherein the increasing step of the thickness of at least a part of the container comprises pressing the container using a thickness adjusting member. 
     
     
         17 . The method for counting counted targets disposed in a container according to  claim 9 , wherein the container comprises a culture container, and the counted targets comprise cells. 
     
     
         18 . A counting apparatus to count counted targets disposed in a liquid enclosed in a container, the counting apparatus comprising:
 a loading base on which the container is placed; and   an adjusting member configured to adjust at least a part of the container including a measurement target region to a predetermined thickness.   
     
     
         19 . The counting apparatus according to  claim 18 , further comprising an agitating member configured to agitate the liquid in the container before the adjusting member adjusts the thickness of the container. 
     
     
         20 . The counting apparatus according to  claim 18 , further comprising:
 photographing means for photographing counted targets disposed in the container;   counting means for counting a number of the counted targets in a photographed image; and
 a driving device configured to, when the number of the counted targets is outside a predetermined range following the counting by the counting means, drive the adjusting member to adjust at least a part of the container to a predetermined thickness so as to render the number of the counted targets in the image within the predetermined range.