The invention provides culture platforms, cell media, and methods of differentiating pluriptent cells into hematopoietic cells. The invention further provides pluripotent stem cell-derived hematopoietic cells generated using the culture platforms and ethods disclosed herein, which enable feed-free, monolayer culturing and in the absence of EB formation. Specifically, pluripotent stem cell-derived hematopoietic cell of this invention include, and not limited to, iHSC, definitive hemogenic endothelium, hematopoietic multipotent progenitors, T cell progenitors, NK cell progenitors, T cells, and NK cells.

The present specification provides a spontaneous-contracting cardiac organoid, a method for manufacturing the organoid, and a method for evaluating drug toxicity by using same, the cardiac organoid comprising: a chamber in which a fluid is stored; a first pipe connected to the chamber so that the fluid flows therethrough; a second pipe connected to the chamber so that the fluid is discharged therethrough; and a valve formed on the first pipe so as to spontaneously open/close an inflow pipe.

A method for producing a cell population containing cardiomyocytes, including (1) a step of bringing a histone deacetylase inhibitor into contact with a cell population containing cardiomyocytes and cells other than cardiomyocytes, the cell population being obtained by culturing pluripotent stem cells in a medium for cardiomyocyte differentiation, and (2) a step of culturing the cell population is provided by the present invention.

The present invention provides a culture method capable of maturing an organoid through long-term culture, and suitable for producing a conformational organ. The culture method of the present invention is a method for culturing an organoid and/or cells constituting an organ immobilized in a chamber with a culture fluid perfused, and the culture fluid is perfused in such a manner as to generate a turbulent flow in the chamber.

A method for generating a heart organoid is provided. The method includes forming a cellular aggregate of pluripotent stem cells, activating Wnt signaling in the cellular aggregate to cause the cellular aggregate to differentiate into a three-dimensional cardiac mesoderm, and inhibiting the Wnt signaling in the cardiac mesoderm to form the heart organoid. The heart organoid includes myocardial tissue, endocardial tissue defining at least one chamber, and epicardial tissue disposed on at least an outer surface of the myocardial tissue. The heart organoid beats. Heart organoids prepared in accordance with the method are also provided.

The present invention provides, in order to prepare matured hepatocytes analogous in various points to primary hepatocytes, a method for preparing hepatocytes or cells that can be differentiated into hepatocytes from pluripotent stem cells, comprising the steps of: (1) culturing the pluripotent stem cells in a medium containing an activator of an activin receptor-like kinase-4,7; (2) culturing the cells obtained in the step (1) in a medium containing a bone morphogenetic factor and a fibroblast growth factor; (3) culturing the cells obtained in the step (2) in a medium containing an activator of a hepatocyte growth factor receptor and an activator of an oncostatin M receptor; and (4) culturing the cells obtained in the step (3) to obtain hepatocytes or cells that can be differentiated into hepatocytes, wherein in at least one of the steps (2), (3) and (4), cells are cultured on a high-density collagen gel membrane.

This disclosure relates generally to the differentiation of human pluripotent stem cells, and more particularly to a method of spatially adsorbing morphogens to differentiate human pluripotent stem cells.

The invention provides a method for producing a cell aggregate containing a ciliary marginal zone-like structure by culturing a cell aggregate containing a retinal tissue in which Chx10 positive cells are present in a proportion of 20% or more of the tissue in a serum-free medium or serum-containing medium, each containing a substance acting on the Wnt signal pathway for only a period before the appearance of a RPE65 gene expressing cell, followed by culturing the “cell aggregate in which a RPE65 gene expressing cell does not appear” thus obtained in a serum-free medium or serum-containing medium, each not containing a substance acting on the Wnt signal pathway and so on.

Disclosed are compositions, in particular, organoid compositions, derived from more than one donor cell. Further disclosed are methods of making compositions, for example, organoid compositions, that comprise a differentiated cell population derived from more than one donor cell. Donor cells may include, for example, a precursor cell such as an embryonic stem cell or other precursor cell. The disclosed methods use synchronization conditions to produce a synchronized pooled-precursor cell population, which may then be differentiated into an organoid composition. Methods of using the compositions are also disclosed.

A factor that is caused by a nucleic acid and influences the kinetics of an extracellular vesicle is screened. A library of barcoded extracellular vesicles is provided.