Culture container for culturing epithelial cells and use thereof

A method for culturing epithelial cells, comprising:

• coating a surface of an upper container of a culture container for culturing epithelial cells with an extracellular matrix;
• accommodating a cell culture medium and the upper container in a lower container;
• inoculating the extracellular matrix with an epithelial cell;
• incubating the culture container for culturing epithelial cells under a culture condition to form a layer of a 2D organoid on the extracellular matrix by the epithelial cell;
• fitting a lid member in an airtight manner to an opening portion of the upper container;
• and
• further incubating the culture container for culturing epithelial cells under the culture condition;
• wherein the culture container comprises:
• an upper container;
• a lid member configured to fit in an airtight manner to an opening portion of the upper container;
• and
• a lower container configured to accommodate the upper container and a cell culture medium,
• wherein at least a part of an area of the upper container in contact with the cell culture medium is formed of a membrane that is permeable to at least a part of the components of the cell culture medium and impermeable to a cell, and
• a material of the lid member has an oxygen permeability coefficient of 1.0×10−6 cm3 cm/(cm2·sec·atm) or less at a pressure of 1 atm at a temperature of 25° C.

A co-culture producing method comprising:

• coating a surface of an upper container of a culture container for culturing epithelial cells with an extracellular matrix;
• accommodating a cell culture medium and the upper container in a lower container;
• inoculating the extracellular matrix with an epithelial cell;
• incubating the culture container for culturing epithelial cells under a culture condition to form a layer of a 2D organoid on the extracellular matrix by the epithelial cell;
• fitting a lid member in an airtight manner to an opening portion of the upper container;
• further incubating the culture container for culturing epithelial cells under the culture condition;
• and
• inoculating the upper container, in which the layer of the 2D organoid is formed, with an anaerobic bacterium, after the incubating the culture container and before the fitting the lid member in an airtight manner;
• wherein the culture container comprises:
• an upper container;
• a lid member configured to fit in an airtight manner to an opening portion of the upper container;
• and
• a lower container configured to accommodate the upper container and a cell culture medium,
• wherein at least a part of an area of the upper container in contact with the cell culture medium is formed of a membrane that is permeable to at least a part of the components of the cell culture medium and impermeable to a cell, and
• a material of the lid member has an oxygen permeability coefficient of 1.0×10−6 cm3 cm/(cm2·sec·atm) or less at a pressure of 1 atm at a temperature of 25° C.