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.

Description

TECHNICAL FIELD[0001]The present invention relates to a method for culturing cells and a cell culturing apparatus that are for culturing cells such as cells, tissues, and microorganisms, and to a counting method and a counting apparatus to count counted targets in a container.BACKGROUND ART[0002]In recent years, demands exist for efficient large-scale cell culturing, tissues, and microorganisms under artificial environments, in fields such as pharmaceutical manufacturing, gene therapy, regenerative medical therapy, and immunotherapy.[0003]In such large-scale cell culturing, especially regarding culturing of floating cells, agitation culturing is commonly practiced using culture vessels equipped with agitating blades. However, agitating blades are not used with cells subject to damage from external force or cells that proliferate while forming aggregates. Instead, a widely employed method is to enclose cells in a culture container where the cells are cultured while being kept station...

AI Technical Summary

Benefits of technology

[0039]With the embodiments of the present invention, aggregates are adjusted to appropriate sizes during cell culturing of cells, tissues, and microorganisms, thereby improving cell proliferation efficiency.

[0040]Also with the embodiments of the present invention, the number of cells is counted without disassembly of a culture system and irrespective of the density of proliferated cells.

Problems solved by technology

However, agitating blades are not used with cells subject to damage from external force or cells that proliferate while forming aggregates.

Unfortunately, the stationary culturing is problematic in that as the aggregates of cells grow larger with the proliferation of cells, this causes a gradual deficiency of oxygen and nutrients to be fed to the cells, leading to degraded proliferation efficiency.

Additionally, although the transfer to another container involves agitation of a culture solution to cancel the unevenness of oxygen and nutrients, another problem arises in that the handling during the transfer causes damage to the cells, leading to degraded proliferation efficiency.

That is, an excessively high density of cells in the culture solution prevents a sufficient supply of oxygen and nutrients to each cell, leading to degraded cell proliferation efficiency.

Likewise, an excessively low density of cells in the culture solution prevents sufficient securement of cell proliferation efficiency.

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