Abstract: A cell culture container includes a storage container, a cell culture area, a culture solution supply unit, and a discharge unit. The cell culture area is a region for culturing cells, and is provided in the storage container. The culture solution supply unit supplies the culture solution into the cell culture area at a flow rate equal to or less than a predetermined flow rate. The discharge unit discharges the liquid in the cell culture area to the outside of the storage container. A rectification unit is a structure in which a plurality of outflow ports are uniformly arranged in a direction perpendicular to the flow direction of the culture solution from the culture solution supply unit to the discharge unit.

Abstract: A revised technique for preparing human tissues and organs aiming at application to medical use is provided. With this technique, function per cell is greatly improved and great reduction in production cost is realized. A method of preparing an organ bud, comprising culturing vascular endothelial cells, mesenchymal cells and a tissue or organ cell in vitro in the presence of blood cells. Also provided are an organ bud prepared by the above method and a method of preparing a tissue or an organ using the organ bud. Using the same organ bud, there are provided a method of transplanting an organ bud, a method of regeneration or function recovery of a tissue or an organ, a method of preparing a non-human chimeric animal, and a method of evaluating a drug.

Abstract: The present invention provides a method for preparing chondrocytes which enable construction of a cartilage tissue, the method being capable of solving the problems with conventional methods. A method for preparing chondrocytes, comprising co-culturing chondrogenic cells with vascular cells. A composition for cartilage regenerative therapy, comprising chondrocytes prepared by the above-described method. A method of screening for drugs effective as pharmaceuticals, comprising using chondrocytes prepared by the above-described method, a cartilage tissue formed from the chondrocytes and/or cells derived from the cartilage tissue. A method for preparing a matrix produced by chondrocytes, comprising using chondrocytes prepared by the above-described method, a cartilage tissue formed from the chondrocytes and/or cells derived from the cartilage tissue. A method for cartilage regeneration, comprising transplanting chondrocytes prepared by the above-described method into an organism to form a cartilage tissue.

Abstract: The present invention provides a technique that serves as a platform for inducing human organ cells at a low cost, stably and in a large quantity. A cell inducible after differentiating pluripotent stem cells and then passaging the resultant cells at least once or more times, which is negative for undifferentiated (pluripotent) cell markers NANOG, OCT4, MYC and LIN28A, negative for endoderm cell markers CXCR4, CER1, HHEX and GATA4, positive for intestinal endoderm cell markers CDX2 and HOXB9, negative for a mesenchymal cell marker brachyury (T), negative for a pancreatic cell marker PDX1, and capable of differentiating into at least a hepatocyte, a pancreatic cell and an intestinal cell. Also provided are methods of preparing and amplifying the above cells; a method of preparing organ cells using the above cells; and a method of constructing a working cell bank for preparing organ cells, comprising cryopreserving the above cells.

Abstract: An operation isolator forms an aseptic space. An incubator is connected to the operation isolator, in which cells are stored and cultured. A storage chamber stores articles used in the operation isolator. In order to carry articles from the outside into the storage chamber, a decontamination pass-box is provided. The storage chamber and the operation isolator are directly or indirectly connected to each other.

Abstract: It is intended to develop a technique that can reproduce a microenvironment of cancer tissue and to construct a novel drug discovery screening system of high precision. It is also intended to provide a method for reconstituting human cancer tissue using primary human cancer cells that retain the properties of human tumor. The present invention provides a reconstituted cancer organoid reproducing a cancer microenvironment. The present invention also provides a method for preparing a cancer organoid from cancer tissue, a xenograft prepared from the cancer organoid, a method for preparing the xenograft, a method for evaluating treatment resistance of cancer, a method for evaluating invasion or metastasis of cancer, a method for evaluating recurrence of cancer, and a method for conducting prognostic prediction of cancer.

Abstract: The present invention provides a culture method for culturing, in recesses (10), a population including two or more cells including a cell derived from a stem cell and a mesenchymal cell. The cell derived from a stem cell is a cell obtained by differentiating a stem cell in vitro. The cell is a cell of one or more types selected from the group consisting of an endodermal cell, an ectodermal cell, and a mesodermal cell. The population is cultured in the recesses (10) together with a vascular cell or a secretor factor. Each recess (10) includes a space in which cells are movable. When a volume of the space is represented by V mm3 and the number of mesenchymal cells seeded in the space is represented by N, V is 400 or less and N/V is in a range from 35 to 3000.

Abstract: The present invention provides a means for reconstituting tissues and organs having mature functions. A method of preparing a tissue or an organ, comprising coculturing an organ cell with a vascular endothelial cell and a mesenchymal cell, generating an organ bud, transplanting the organ bud into a non-human animal, and then isolating from the non-human animal the transplanted organ bud-derived tissue or organ.

Abstract: The present invention finds out find out the requirements necessary for preparing a cell condensate in vitro from a large number of cells (several ten thousand to several million cells) and provides a method of forming a cell condensate for self-organization which is capable of realizing complex higher structures (such as liver and kidney) and interactions with other organs. A method of preparing a cell condensate in vitro, comprising culturing a mixture of cells and/or tissues of a desired type in a total cell count of 400,000 or more and 100,000 to 400,000 mesenchymal cells to form a cell condensate of 1 mm or more in size. A cell condensate prepared by the above-described method. A method of preparing a three-dimensional tissue structure, comprising allowing self-organization of a cell condensate prepared by the above-described method to form a three-dimensional tissue structure that has been provided with higher structures.

Abstract: The present invention provides a method of constituting a tissue construct in vitro using a tissue without depending on scaffold materials. A method of integrating a biological tissue with a vascular system in vitro, comprising coculturing a biological tissue with vascular cells and mesenchymal cells. A biological tissue which has been integrated with a vascular system by the above-described method. A method of preparing a tissue or an organ, comprising transplanting the biological tissue described above into a non-human animal and differentiating the biological tissue into a tissue or an organ in which vascular networks have been constructed. A method of regeneration or function recovery of a tissue or an organ, comprising transplanting the biological tissue described above into a human or a non-human animal and differentiating the biological tissue into a tissue or an organ in which vascular networks have been constructed.

Abstract: A culture chamber includes a plurality of recesses (10) each formed of a bottom portion (11) and an opening portion (12). The bottom portion (11) has a hemispherical shape and the opening portion (12) is defined by a wall that surrounds an area from a boundary between the opening portion (12) and the bottom portion (11) to an end of each of the recesses (10), the wall having a taper angle in a range from 1 degree to 20 degrees. An equivalent diameter of the boundary is in a range from 50 ?m to 2 mm and a depth from a bottom of the bottom portion (11) to the end of each of the recesses is in a range from 0.6 or more times to 3 or less times the equivalent diameter, and the wall defining the opening portion (12) forms a surface continuous to the bottom portion (11).

Abstract: A tissue structure for enabling comprehensive understanding of gene patterns of mature cells and a method of preparing the tissue structure are provided. A tissue structure is obtained by co-culturing an endodermal, ectodermal, or mesodermal cell derived from a stem cell and at least one cell and/or factor selected from the group consisting of a vascular cell, a mesenchymal cell, a factor secreted by a vascular cell, a factor secreted by a mesenchymal cell, and a factor secreted when both a vascular cell and a mesenchymal cell exist. A value obtained by assay of a plurality of functions using a Pearson product-moment correlation coefficient is closer to a value of a cell or biological tissue sampled from an adult than a value of a cell or biological tissue sampled from a fetus.

Abstract: The present invention provides a method for preparing chondrocytes which enable construction of a cartilage tissue, the method being capable of solving the problems with conventional methods. A method for preparing chondrocytes, comprising co-culturing chondrogenic cells with vascular cells. A composition for cartilage regenerative therapy, comprising chondrocytes prepared by the above-described method. A method of screening for drugs effective as pharmaceuticals, comprising using chondrocytes prepared by the above-described method, a cartilage tissue formed from the chondrocytes and/or cells derived from the cartilage tissue. A method for preparing a matrix produced by chondrocytes, comprising using chondrocytes prepared by the above-described method, a cartilage tissue formed from the chondrocytes and/or cells derived from the cartilage tissue. A method for cartilage regeneration, comprising transplanting chondrocytes prepared by the above-described method into an organism to form a cartilage tissue.

Abstract: The present invention provides a means for reconstituting tissues and organs having mature functions. A method of preparing a tissue or an organ, comprising coculturing an organ cell with a vascular endothelial cell and a mesenchymal cell, generating an organ bud, transplanting the organ bud into a non-human animal, and then isolating from the non-human animal the transplanted organ bud-derived tissue or organ.