Abstract: A method for observing the dynamics of sweat glands and a method for evaluating a substance of interest, which are useful for development of a preparation for external application, such as a cosmetic. In each of the methods, an observation sample is used, which is prepared by staining all sweat glands, which are isolated alive, with a staining reagent, and then holding the all sweat glands on a support using at least one material selected from the group consisting of collagen, agarose, basement membrane matrix, poly-D-lysine and a membrane.

Abstract: Provided is a method for producing vascular endothelial cells from pluripotent stem cells, the method comprising the following steps (i) to (iii): (i) a step of culturing pluripotent stem cells in a culture medium comprising a BMP, on a culture vessel coated with a first matrix, to produce mesodermal progenitor cells; (ii) a step of dissociating the resulting cells into single cells; and (iii) a step of culturing the resulting cells in a culture medium comprising VEGF, on a culture vessel coated with a second matrix selected from the group consisting of laminin-411 or a fragment thereof, laminin-511 or a fragment thereof, Matrigel, type IV collagen and fibronectin.

Abstract: A modified laminin characterized in that a laminin or a heterotrimeric laminin fragment has a collagen binding molecule conjugated to at least one site selected from the ? chain N-terminus, the ? chain N-terminus and the ? chain N-terminus, and an extracellular-matrix material comprising the modified laminin, and collagen and/or gelatin serve as an alternative to Matrigel and are useful as an extracellular-matrix material for the formation of a safe three-dimensional tissue structure for regenerative medicine in humans.

Abstract: Provided are a modified laminin having a cell-growth regulatory molecule bound to at least one site selected from the ? chain N-terminus, the ? chain C-terminus, the ? chain N-terminus and the ? chain N-terminus of laminin or a heterotrimeric laminin fragment, a method for culturing cells in the presence of the modified laminin, a method for establishing iPS cells in the presence of the modified laminin, and a culture substrate coated with the modified laminin. Human stem cells cultured in a xeno-free environment with the use of the modified laminin of the present invention can be provided as highly safe human stem cells applicable to regenerative medicine.

Abstract: The present invention provides a novel technique in a cell culture method using a cell culture vessel coated with a laminin fragment, which novel technique achieves cell culture as in the case of using a recommended coating concentration even when the coating concentration is lower than the recommended coating concentration. The present invention relates to a method for enhancing an activity for mammalian cultured cells of a laminin fragment or a variant thereof each having integrin binding activity.

Abstract: The present invention provides a method of purifying highly pure retinal pigment epithelial cells from a cell population obtained by induction of differentiation of pluripotent stem cells into retinal pigment epithelial cells, by a simple and easy operation in a short period. The purification method of the present invention includes a step of introducing a cell population containing retinal pigment epithelial cells obtained by differentiation induction of pluripotent stem cells on laminin or a fragment thereof on a filter, and obtaining a cell population that passed the filter.

Abstract: Provided are a production method of retinal pigment epithelial (RPE) cells that improves differentiation induction efficiency of pluripotent stem cells into RPE cells, and can provide highly pure RPE cells by a simple and easy operation in a short period, a culture method of RPE cells that can stably grow and culture a cell, a toxicity/efficacy evaluation method using RPE cells useful for transplantation therapy, and a therapeutic drug for a retinal disease. The invention relates to a production method of RPE cells, comprising adhesion culture of human pluripotent stem cells using a culture substrate coated with a laminin-E8 fragment, a culture method of RPE cells, comprising adhesion culture of RPE cells using a culture substrate coated with a laminin-E8 fragment, a toxicity or efficacy evaluation method using RPE cells obtained by producing or culturing by the method, and a therapeutic drug for a retinal disease, containing the RPE cells.

Abstract: The present invention provides a method for producing dopaminergic neuron progenitor cells from pluripotent stem cells, which method comprises the steps of: (i) performing adherent culture of pluripotent stem cells on an extracellular matrix in a medium containing a reagent(s) selected from the group consisting of BMP inhibitor, TGF? inhibitor, SHH signal-stimulating agent, FGF8, and GSK3? inhibitor; (ii) collecting Corin- and/or Lrtm1-positive cells from the cells obtained in Step (i) using a substance which binds to Corin and/or a substance which binds to Lrtm1; and (iii) performing suspension culture of the cells obtained in Step (ii) in a medium containing a neurotrophic factor.

Abstract: Provided is a cell culture vessel characterized in that a surface to be in contact with cells is coated with a laminin fragment having integrin ?6?1 binding activity or a modified form thereof in a dry state, the laminin fragment being derived from at least one kind selected from laminin ?5?1?1 and laminin ?5?2?1, the cell culture vessel being any of the following: (1) a cell culture vessel of which a surface to be in contact with cells is coated only with a laminin fragment having integrin ?6?1 binding activity or a modified form thereof in a dry state; (2) a cell culture vessel of which a surface to be in contact with cells is coated with a laminin fragment having integrin ?6?1 binding activity or a modified form thereof in combination with a laminin fragment having no integrin ?6?1 binding activity in a dry state; and (3) a cell culture vessel of which a surface to be in contact with cells is coated with a laminin fragment having integrin ?6?1 binding activity or a modified form thereof in combination

Abstract: A modified laminin characterized in that a laminin or a heterotrimeric laminin fragment has a collagen binding molecule conjugated to at least one site selected from the ? chain N-terminus, the ? chain N-terminus and the ? chain N-terminus, and an extracellular-matrix material comprising the modified laminin, and collagen and/or gelatin serve as an alternative to Matrigel and are useful as an extracellular-matrix material for the formation of a safe three-dimensional tissue structure for regenerative medicine in humans.

Abstract: Provided is a method of stably maintaining and culturing “hepatoblast-like cells” generated during a differentiation-inducing process from pluripotent stem cells to hepatocytes. The present invention also provides a culture product obtained by the culture method. The hepatoblast-like cells can be maintained and cultured stably by bringing a laminin into contact with the hepatoblast-like cells. The method of the present invention which uses a laminin makes it possible for the first time to culture, maintain, and proliferate the hepatoblast-like cells. Desired mature cells such as mature hepatocytes and bile duct epithelial cells can be generated in a short time period and can be acquired at a desired timing by maintaining the hepatoblast-like cells. Further, the resulting cultured hepatoblast-like cells were demonstrated to be capable of being induced to differentiate into mature hepatocytes, mature cholangiocytes, bile duct epithelial cells, and the like.

Abstract: Provided is a novel ligand for integrin ?9?1 consisting of a peptide having the following amino acid sequence: (A) EDDMMEVPY (SEQ ID NO: 1) or (B) an amino acid sequence the same as the amino acid sequence represented by SEQ ID NO: 1 except for having deletion, substitution or addition of 1 to 3 amino acids. The novel ligand for integrin ?9?1 has a higher binding affinity than those of tenascin-C and osteodontin, which are known ligands for integrin ?9?1.

Abstract: The present invention provides a culture substrate which enables maintenance culture of human pluripotent stem cells in a pluripotent state under a feeder-free culture environment, and a culture method of human pluripotent stem cells using the culture substrate. By seeding human pluripotent stem cells dissociated into single cells at a cell density of 4×104 to 10×104 cells/cm2 onto a culture substrate coated with human laminin ?5?1?1 E8 fragment or human laminin ?3?3?2 E8 fragment preferably at a concentration of 0.5 to 25 ?g/cm2, the human pluripotent stem cells can be rapidly expanded in a pluripotent state.

Abstract: Provided are a modified laminin having a cell-growth regulatory molecule bound to at least one site selected from the ? chain N-terminus, the ? chain C-terminus, the ? chain N-terminus and the ? chain N-terminus of laminin or a heterotrimeric laminin fragment, a method for culturing cells in the presence of the modified laminin, a method for establishing iPS cells in the presence of the modified laminin, and a culture substrate coated with the modified laminin. Human stem cells cultured in a xeno-free environment with the use of the modified laminin of the present invention can be provided as highly safe human stem cells applicable to regenerative medicine.

Abstract: It is an object of the present invention to provide a Type I-Type IV collagen hybrid gel, which maintains characteristics of a Type IV collagen and is superior in gel strength. It is the Type I-Type IV collagen hybrid gel obtained by mixing 100 to 500 parts by mass of the Type I collagen having fibrosis ability, relative to 100 parts by mass of the Type IV collagen having gelling ability. A three-dimensional structure is formed, where a membrane-like substance by the Type IV collagen is formed onto a fibrous substance by the Type I collagen, so as to be able to provide cell culture environment approximate to a basement membrane of a living body.

Abstract: The present invention provides a culture substrate which enables maintenance culture of human pluripotent stem cells in a pluripotent state under a feeder-free culture environment, and a culture method of human pluripotent stem cells using the culture substrate. By seeding human pluripotent stem cells dissociated into single cells at a cell density of 4×104 to 10×104 cells/cm2 onto a culture substrate coated with human laminin ?5?1?1 E8 fragment or human laminin ?3?3?2 E8 fragment preferably at a concentration of 0.5 to 25 ?g/cm2, the human pluripotent stem cells can be rapidly expanded in a pluripotent state.

Abstract: It is an object of the present invention to provide a Type I-Type IV collagen hybrid gel, which maintains characteristics of a Type IV collagen and is superior in gel strength. It is the Type I-Type IV collagen hybrid gel obtained by mixing 100 to 500 parts by mass of the Type I collagen having fibrosis ability, relative to 100 parts by mass of the Type IV collagen having gelling ability. A three-dimensional structure is formed, where a membrane-like substance by the Type IV collagen is formed onto a fibrous substance by the Type I collagen, so as to be able to provide cell culture environment approximate to a basement membrane of a living body.

Abstract: A method of determining a laminin 5 antigen in a biological sample, comprising the steps of bringing an antibody reactive to a laminin 5 ?2 chain N-terminal fragment into contact with the biological sample; measuring a reaction of the antibody; and determining an amount of the laminin 5 antigen based on a measurement result of the reaction, as well as, a method of detecting a laminin 5-producing tumor cell, a method of examining acute respiratory distress syndrome and a method of evaluating malignancy of a malignant tumor based on the assay method.

Abstract: A method of determining a laminin 5 antigen in a biological sample, comprising the steps of bringing an antibody reactive to a laminin 5 ?2 chain N-terminal fragment into contact with the biological sample; measuring a reaction of the antibody; and determining an amount of the laminin 5 antigen based on a measurement result of the reaction, as well as, a method of detecting a laminin 5-producing tumor cell, a method of examining acute respiratory distress syndrome and a method of evaluating malignancy of a malignant tumor based on the assay method.

Abstract: A method of determining a laminin 5 antigen in a biological sample, comprising the steps of bringing an antibody reactive to a laminin 5 &ggr;2 chain N-terminal fragment into contact with the biological sample; measuring a reaction of the antibody; and determining an amount of the laminin 5 antigen based on a measurement result of the reaction, as well as, a method of detecting a laminin 5-producing tumor cell, a method of examining acute respiratory distress syndrome and a method of evaluating malignancy of a malignant tumor based on the assay method.