Means which enables preparation of a thick cell aggregate by a simple process without an operation of detaching and stacking of cells is disclosed. The method for preparing a three-dimensional cell aggregate by the present invention comprises: a cell encasing step of placing a cell suspension in a cell container; and a pressure application step of applying pressure to cells in the container. The cell encasing step and the pressure application step may be carried out a plurality of times. By the present invention, a thick, robust cell aggregate can be obtained by a simple operation of applying pressure to a cell suspension or a medium containing cells.

Since the method does not require an operation of stacking a plurality of cell sheets, the cells are hardly damaged, and the conditions of the cells can be favorably maintained, so that the cells can be advantageously used as a tissue piece for transplantation.

The tissue regeneration platform of one embodiment may exhibit excellent biocompatibility and tissue culture property on various supports, comprising a support and a coating layer disposed on the support, and comprising a hyaluronic acid-catechol compound.

A method for enhancing activity selected from the group consisting of cytokine production capacity, proliferation capacity, engraftment capacity, angiogenesis-inducing capacity, and tissue regeneration capacity in a graft, particularly in a sheet-shaped cell culture containing a somatic cell, involves incubating the graft at a temperature of 25° C. or higher.

Methods of using human airway stem cells in lung epithelial engineering, optionally wherein the cells are contacted with a gamma secretase inhibitor, bioartificial airway organs produced thereby, and the use thereof, e.g., for transplantation. Also methods of treating a bio-artificial matrix with Tenascin-C and/or fibrillin 2.

The invention provides a method of producing a connective tissue graft suitable for correcting a connective tissue defect, comprising determining the size and shape of a tissue defect, obtaining a fat tissue from a patient modelled to fit the size and shape of the tissue defect, contacting the fat tissue with one or more connective tissue specific growth or differentiation factors; and kits for such a method.

The invention provides a method for producing a ciliary marginal zone stem cell induced to differentiate from a pluripotent stem cell, including either the following step (1) or step (2), or both of these steps: (1) a step of floating culturing cells obtained from a cell aggregate containing a ciliary marginal zone-like structure induced to differentiate from pluripotent stem cells, thereby obtaining a retinosphere; and (2) a step of collecting stage specific embryonic antigen-1 positive cells from cells obtained from a cell aggregate containing a ciliary marginal zone-like structure induced to differentiate from pluripotent stem cells.

The present invention provides a cell culture substrate containing a nanofiber composed of a biodegradable polymer on a support composed of a biodegradable polymer. It also provides a method of culturing cells, which includes seeding cells on the substrate, and stationary culture of the cells. Furthermore, the present invention provides an agent for cell transplantation therapy, which contains the substrate and cells cultured on the substrate.

There is disclosed a method of combining mesenchymal stem cells (MSCs) with a bone substrate. In an embodiment, the method includes obtaining tissue having MSCs together with unwanted cells. The tissue is digested to form a cell suspension having MSCs and unwanted cells. The cell suspension is added to the substrate. The substrate is cultured to allow the MSCs to adhere. The substrate is rinsed to remove unwanted cells. In various embodiments, the tissue is adipose tissue, muscle tissue, or bone marrow tissue. In an embodiment, there is disclosed an allograft product including a combination of MSCs with a bone substrate in which the combination is manufactured by culturing MSCs disposed on the substrate for a period of time to allow the MSCs to adhere to the substrate, and then rinsing the substrate to remove unwanted cells from the substrate. Other embodiments are also disclosed.

A bioreactor for preferably three-dimensional cell culturing comprises a scaffold chamber, a first tube, a second tube and a first valve with a scaffold adapter, a tube adapter and a medium adapter. The first valve has a housing with a longitudinal female portion ending in an opening and a longitudinal actuator being arranged through the opening of the female portion of the housing such that the actuator is arranged partially inside the housing and partially outside the housing, wherein the actuator of the first valve is axially moveable relative to the housing of the first valve between a first position in which the first valve is in the operation position and a second position in which the first valve is in the medium change position. By providing the actuators in the first valve which is applied by axial movements, operation of the bioreactor can be comparably simple and safe.

The present invention provides: a method for producing mixed cells comprising cardiomyocytes, endothelial cells and mural cells from induced pluripotent stem cells, the method comprising (a) a step of producing cardiomyocytes from induced pluripotent stem cells and (b) a step of culturing the cardiomyocytes in the presence of VEGF; and a therapeutic agent for heart diseases, comprising the mixed cells produced by the method.