Low oxygen tension is a critical regulator of the developing or regenerating vasculature. The present invention is based on the determination that low oxygen tension during early stages of early vascular cell (EVC) derivation induces endothelial commitment and maturation of pluripotent stem cells. Inhibition of reactive oxygen species generation during the early stages of differentiation abrogates the endothelial inductive effects of the low oxygen environments. Methods of generating various types of cells from pluripotent stem cells (PSCs) are described, as well as compositions and methods of use thereof. In particular, generation of EVCs, bicellular vascular populations, early endothelial cells (ECs) and pericytes via culture in a low oxygen environment is described.

Low oxygen tension is a critical regulator of the developing or regenerating vasculature. The present invention is based on the determination that low oxygen tension during early stages of early vascular cell (EVC) derivation induces endothelial commitment and maturation of pluripotent stem cells. Inhibition of reactive oxygen species generation during the early stages of differentiation abrogates the endothelial inductive effects of the low oxygen environments. Methods of generating various types of cells from pluripotent stem cells (PSCs) are described, as well as compositions and methods of use thereof. In particular, generation of EVCs, bicellular vascular populations, early endothelial cells (ECs) and pericytes via culture in a low oxygen environment is described.

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.

The present invention provides methods for cellular seeding onto three-dimensional fibroblast constructs, three-dimensional fibroblast constructs seeded with muscle cells, and uses therefore.

The present invention provides methods for cellular seeding onto three-dimensional fibroblast constructs, three-dimensional fibroblast constructs seeded with muscle cells, and uses therefore.

A method of differentiating pluripotent stem cells into hemangioblasts comprising incubating the pluripotent stem cells in a first serum-free differentiation medium comprising bone morphogenetic protein 4 (BMP4), vascular endothelial growth factor (VEGF) and stem cell factor (SCF) to induce differentiation of the pluripotent stem cells into hemangioblasts or hemangioblast-containing embryoid bodies is provided. The hemangioblasts or embryoid bodies may be cultured in a second differentiation medium comprising at el least granulocyte-macrophage colony stimulating factor (GM-CSF), macrophage colony stimulating factor (M-CSF) and interleukin-3 (IL-3) for a period of time sufficient to generate alveolar-like macrophages.

In an embodiment, an object of the present invention is to provide a therapeutic and/or preventive agent for a fibrosis such as pulmonary fibrosis, an inflammatory disease and/or an aging disease. In an embodiment, the present invention relates to a therapeutic and/or preventive agent for at least one disease of a fibrosis, an inflammatory disease and an aging disease, comprising extracellular vesicle derived from cell of a tissue or a surrounding tissue thereof where disease can occur, in which the cell is differentiated cell, and relates to a pharmaceutical composition containing the therapeutic and/or preventive agent.

This application relates to a method of making a cell construct, comprising a) plating a plurality of cells on a substantially flat surface; b) growing the plurality of cells to at least 80% confluent to form a cell sheet with intercellular linkages; c) applying a culture medium having a pH of about 5 to about 6.8 to the cell sheet; d) replacing the culture medium of step c) with a culture medium having a pH of about 7.5 to about 8.5; and e) replacing the culture medium of step d) with a culture medium having a pH of about 7 to about 7.7, to obtain a substantially planar untethered cell sheet. Also provided is a cell construct formed according to the method and uses thereof.

This application relates to a method of making a cell construct, comprising a) plating a plurality of cells on a substantially flat surface; b) growing the plurality of cells to at least 80% confluent to form a cell sheet with intercellular linkages; c) applying a culture medium having a pH of about 5 to about 6.8 to the cell sheet; d) replacing the culture medium of step c) with a culture medium having a pH of about 7.5 to about 8.5; and e) replacing the culture medium of step d) with a culture medium having a pH of about 7 to about 7.7, to obtain a substantially planar untethered cell sheet. Also provided is a cell construct formed according to the method and uses thereof.

In one aspect, a method of preconditioning stem cells comprising exposing stem cells to low dose radiation (LDR) is provided. In another aspect, a population of preconditioned stem cells is provided, wherein the population of 5 preconditioned stem cells is obtained by exposing stem cells to LDR. Uses of the preconditioned stem cells are also provided. In other aspects, the stem cells are muscle stem cells.