Osaka University

Kyoto University

Eisai R&d Management Co., Ltd.

Mandom Corporation

Healios K.k.

Japan Institute of Leather Research

Kiyotoshi Sekiguchi

Megumu Saito

Ryuichiro Kurata

Fumitaka Fujita

Sugiko Futaki

Ko Tsutsui

Ryoko Sato

Yukimasa Taniguchi

Kohji Nishida

Yoshiki Sawa

Yuichi Ono

Norio Nakatsuji

Jun Takahashi

Shigeru Miyagawa

Tatsutoshi Nakahata

Shunji Hattori

Ryuhei Hayashi

Masanori Sawada

Akira Niwa

Masato Nakagawa

Hirofumi Suemori

Fumihiro Okada

Daisuke Doi

Yoh-ichi Koyama

Eihachiro Kawase

Hidetoshi Sakurai

Masahiko Katayama

Bumpei Samata

Takamichi Miyazaki

Shun Shibata

Ichiro Katayama

Maria Hayashi

Hitomi Fujisaki

Noriko Sanzen

Mingming Zhao

Fumi Ebisu

Antti Markus Siltanen

Hiroyuki Murota

Ryo Ota

Yukiko Ochi

Atsushi Tanemura

Shaoliang Li

Kie Nakashima

Tomohisa Hayakawa

Provided is a method for inducing pluripotent stem cells to differentiate into somatic cells in a culture medium containing a heparin binding growth factor, the method comprising bringing cells into contact with a conjugate of a laminin E8 fragment and a growth factor binding domain-containing fragment of a heparan sulfate proteoglycan. According to the present invention, pluripotent stem cells can be induced to differentiate into any desired somatic cells in a highly efficient manner.

Method for inducing pluripotent stem cells to differentiate into somatic cells

The present invention relates to a method for controlling differentiation of pluripotent stem cells, which method comprises selecting a laminin or a fragment thereof based on binding affinity for the pluripotent stem cells and inducing differentiation of the pluripotent stem cells in the presence of the laminin or a fragment thereof. Here, the binding affinity for cells can be assessed by time-course observation of the survival rate and motility of the cells. According to the present invention, a cell population containing any desired proportion of differentiated cells can be produced from pluripotent stem cells in a simple manner. The cell population obtained by this production method is very useful for cell therapy-based treatment strategies for diseases.

Method for controlling differentiation of pluripotent stem cells

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.

Method for purification of retinal pigment epithelial cells

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&#x3b2; inhibitor, SHH signal-stimulating agent, FGF8, and GSK3&#x3b2; 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.

Method for inducing dopaminergic neuron progenitor cells

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.

Method for observing dynamics of sweat glands

An immortalized sweat gland myoepithelial cell which expresses &#x3b1;-SMA and pan-cytokeratin and has a sphere forming ability after subculture at least 5 times. A method for producing immortalized sweat gland myoepithelial said method comprising: while culturing a cell structure, wherein sweat gland myoepithelial cells are exposed on a surface, in a state of being suspended in a medium, transferring an immortalizing gene into the cells; and then culturing the transgenic structure thus obtained in a state of being suspended in the medium to thereby obtain immortalized sweat gland myoepithelial cells.

Immortalized sweat gland myoepithelial cell

The present invention provides a method for preparing a clinically applicable, safe and less damaged cardiomyocyte population through a brief and simple procedure from a cell population obtained by induced differentiation of pluripotent stem cells into cardiomyocytes. The present invention relates to a method for preparing a cardiomyocyte population, the method comprising the steps of:

Method for preparing pluripotent stem cell-derived cardiomyocyte population

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.

Method for inducing vascular endothelial cells

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.

Method for enhancing activity of laminin fragments as cell culture matrix

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 &#x3b1;6&#x3b2;1 binding activity or a modified form thereof in a dry state,

Cell culture vessel coated with laminin fragment in dry state

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 &#x3b1; chain N-terminus, the &#x3b2; chain N-terminus and the &#x3b3; 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.

Modified laminin containing collagen binding molecule and use thereof

Provided are a modified laminin having a cell-growth regulatory molecule bound to at least one site selected from the &#x3b1; chain N-terminus, the &#x3b1; chain C-terminus, the &#x3b2; chain N-terminus and the &#x3b3; 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.

Modified laminin and use thereof

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&#xd7;10to 10&#xd7;10cells/cmonto a culture substrate coated with human laminin &#x3b1;5&#x3b2;1&#x3b3;1 E8 fragment or human laminin &#x3b1;3&#x3b2;3&#x3b3;2 E8 fragment preferably at a concentration of 0.5 to 25 &#x3bc;g/cm, the human pluripotent stem cells can be rapidly expanded in a pluripotent state.

Culture substrate for human pluripotent stem cells and use thereof

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.

Type I-type IV collagen hybrid gel

A method of determining a laminin 5 antigen in a biological sample, comprising the steps of bringing an antibody reactive to a laminin 5 &#x3b3;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.

Reagent for determining laminin 5 antigen in biological sample and assay method

Growing community of inventors

Osaka, Japan

Kiyotoshi Sekiguchi