Public University Corporation Yokohama City University

Corning Incorporated

Nikon Corporation

Shimadzu Corporation

Takeda Pharmaceutical Company Limited

Osaka University

Shibuya Corporation

Yokohama City University

Biomedica Solutions Co., Ltd.

Takanori Takebe

Hideki Taniguchi

Yoko Ejiri

Naoto Fukuhara

Satoru Ayano

Keisuke Sekine

Shinji Kobayashi

Yasujiro Kiyota

Masaki Kanai

Hirohisa Abe

Masahiro Kinooka

Masafumi Yamashita

Tetsuya Nishimura

Tomoki Ohkubo

Atsushi Nakao

Yun-Wen Zheng

Akihide Ryo

Hironobu Sunayama

Goro Kobayashi

Hiroshi Yoshikawa

Momotaro Ishikawa

Norikazu Saiki

Yoshinobu Takahashi

Ran Ran Zhang

Takashi Hiromatsu

Yunzhong Nie

Soichiro Murata

Kei Miyakawa

The present invention provides means for producing an organoid close to an organ in a living body and capable of secretion of a plasma protein and immune response. A matrix composition of the present invention provided as such means includes: (1) a first matrix containing one or more cells selected from the group consisting of vascular cells, nerve cells, and blood cells; and (2) a second matrix containing to cells constituting an organ and/or an organoid, in which the first matrix envelops the second matrix, and the first matrix has at least one opening.

Matrix composition

Provided is a culture chamber capable of preparing spheroids with a uniform size with high efficiency and having a micro-space structure which is designed to facilitate replacement of a medium and harvesting of cells. The culture chamber includes a plurality of recesses () each formed of a bottom portion () and an opening portion (). The bottom portion () has one of a hemispherical shape and a truncated cone shape and the opening portion () is defined by a wall that surrounds an area from a boundary between the opening portion () and the bottom portion () to an end of each of the recesses (), 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 μm to 2 mm and a depth from a bottom of the bottom portion () 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 () forms a surface continuous to the bottom portion () and an inclination of the continuous surface changes at the boundary.

Culture chamber and culture method

A cell evaluation method includes acquiring a first evaluation index and a first index calculated using the first evaluation index with respect to comparative target cells in a culture process including a cell differentiation-inducing process in which cell differentiation is induced, calculating a second index on the basis of the first evaluation index with respect to evaluation target cells different from the comparative target cells, and evaluating differentiation of the evaluation target cells by comparing the first index with the second index.

Cell evaluation method, cell evaluation system and program

The present invention solves the following problems [1] to [3] found in conventional methods of preparing a three dimensional structure (organ primordium) by coculturing functional cells with umbilical cord-derived vascular endothelial cells and bone marrow-derived mesenchymal cells: [1] the quality of resultant organ primordia varies greatly depending on donors; [2] the growth capacities of cell sources are limited; and [3] it is difficult to secure immunocompatibility because cells are derived from different sources. An organ bud prepared from vascular cells, mesenchymal cells and tissue or organ cells, wherein each of the vascular cell, the mesenchymal cell and the tissue or organ cell has been induced from pluripotent stem cells. A method of preparing an organ bud, comprising culturing vascular cells, mesenchymal cells and tissue or organ cells in vitro, wherein each of the vascular cell, the mesenchymal cell and the tissue or organ cell has been induced from pluripotent stem cells.

Formation of three-dimensional organ from pluripotent stem cells

Provided is a culture chamber that includes a plurality of recesses each formed of a bottom portion and an opening portion. The bottom portion has a hemispherical shape or a truncated cone shape. The opening portion is defined by a wall that surrounds an area from a boundary between the opening portion and the bottom portion to an end of each of the recesses, the wall having a taper angle in a range from 1 to 20 degrees. An equivalent diameter of the boundary is from 50 μm to 2 mm and a depth from a bottom of the bottom portion to the end of each of the recesses is from 0.6 or more times to 3 or less times the equivalent diameter, and the wall defining the opening portion forms a surface continuous to the bottom portion. An inclination of the continuous surface changes at the boundary.

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. A method of preparing a non-human chimeric animal, comprising transplanting the biological tissue described above into a non-human animal and differentiating the biological tissue into a tissue or organ in which vascular networks have been constructed. A method of evaluating a drug, comprising using at least one member selected from the group consisting of the biological tissue described above, the tissue or organ prepared by the method described above, and the non-human chimeric animal prepared by the method described above. A composition for regenerative medicine, comprising a biological tissue which has been integrated with a vascular system by the method described above.

Method for integrating biological tissues with a vascular system

Provided is a culture chamber that includes a plurality of recesses each formed of a bottom portion and an opening portion. The bottom portion has a hemispherical shape or a truncated cone shape. The opening portion is defined by a wall that surrounds an area from a boundary between the opening portion and the bottom portion to an end of each of the recesses, the wall having a taper angle in a range from 1 to 20 degrees. An equivalent diameter of the boundary is from 50 &#x3bc;m to 2 mm and a depth from a bottom of the bottom portion to the end of each of the recesses is from 0.6 or more times to 3 or less times the equivalent diameter, and the wall defining the opening portion forms a surface continuous to the bottom portion. An inclination of the continuous surface changes at the boundary.

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.

Method for producing tissue and organ

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.

Cell culture vessel

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.

Tissue structure and preparation method thereof

The present invention provides a culture method for culturing, in recesses (), 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 () together with a vascular cell or a secretor factor. Each recess () includes a space in which cells are movable. When a volume of the space is represented by V mmand 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.

Culture method and cell cluster

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.

Cell culture system

A culture chamber includes a plurality of recesses () each formed of a bottom portion () and an opening portion (). The bottom portion () has a hemispherical shape and the opening portion () is defined by a wall that surrounds an area from a boundary between the opening portion () and the bottom portion () to an end of each of the recesses (), 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 &#x3bc;m to 2 mm and a depth from a bottom of the bottom portion () 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 () forms a surface continuous to the bottom portion ().

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.

Primitive gut endoderm cells and method for producing same

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.

Growing community of inventors

Yokohama, Japan

Takanori Takebe