The present invention relates to novel compositions and methods to produce 3D organ equivalents of the brain (i.e. mini-brains). The invention also relates to methods of using human induced pluripotent stem cells, a combination of growth and other soluble factors and gyratory shaking. Cells from healthy or diseased donors or animals can be used to allow testing different genetic backgrounds. The model can be further enhanced by using genetically modified cells, adding micro-glia or their precursors or indicator cells (e.g. with reporter genes or tracers) as well as adding endothelial cells to form a blood-brain-barrier.

Described herein are tissue cultures and methods for culturing stem-cell derived neurons in three-dimensional culture. Also provided are methods for screening therapeutic agents, developing cell line models, identifying genes associated with a neurological disease, and identifying genes associated with neuronal cell phenotypes.

A three dimensional (3D) model of a tumor made of a synthetic material and a plurality of cell types, including malignant cells and non-malignant cells of the tumor, having a full HLA match, such that the synthetic material and the plurality of cell types are arranged in high matchability to a 3D image of the tumor, is provided. Methods of forming the 3D tumor model by bioprinting are also provided, as well as systems in which the 3D tumor model can be perfused and fluidly connected to a medium containing immune cells and/or other cells and factors present in the tumor's microenvironment. Methods utilizing the 3D tumor model or the system in, for example, personalized therapy, are also provided.

Functional, brain region-specific neural spheroids comprising neuronal cells and optionally glial cells at varying ratios are disclosed, as are methods of making such spheroids and methods for their use, such as for modeling particular brain regions that may be implicated in diseases, or for observing drug effects.

A three dimensional (3D) model of a glioblastoma tumor made of a synthetic material and a plurality of cell types, including malignant cells and non-malignant cells of the tumor. Methods of forming the 3D tumor model are also provided, as well as systems in which the 3D tumor model can be perfused and fluidly connected to a medium containing immune cells and/or other cells and factors present in the tumor's microenvironment. Methods utilizing the 3D tumor model or the system in, for example, personalized therapy, are also provided.

Compositions and methods are provided for biologically relevant in vitro screening of neural function, including determination of the effects of an agent on neural cells. The compositions of the invention useful in such screening methods include a neural co-culture system comprising human pluripotent stem cell (PSC)-derived neurons and human glial cells, which may be derived by culture methods allowing for rapid and robust development of highly mature neuronal activity, particularly spontaneous synchronous network bursts.

The purpose of the present invention is to efficiently produce microglia from pluripotent stem cells. Provided is a method for producing microglia from pluripotent stem cells, comprising the following steps: (a) a step of co-culturing a pluripotent stem cell together with a feeder cell for 7 days or longer, and obtaining a blood progenitor cell; (b) a step of co-culturing the blood progenitor cell obtained in step (a) together with a feeder cell in the presence of IL-3 and/or GM-CSF, and obtaining an embryonic monocyte; and (c) a step of, in the presence of M-CSF, co-culturing the embryonic monocyte obtained in step (b) together with an astrocyte, or culturing the embryonic monocyte using an astrocyte supernatant.

Provided herein is an in vitro model of the blood brain barrier. In some embodiments, the model includes: an endothelial cell layer, and brain tissue layer comprising neuronal cells, and optionally one or more of astrocytes, pericytes, oligodendrocytes, and microglia. In some embodiments, the model further comprises a porous membrane between said endothelial cell layer and the neuronal cell layer. A microfluidic device comprising the same and methods of use thereof are also provided.

The present invention relates to: a cell therapeutic agent including ventral midbrain-derived astrocytes and dopamine neural progenitor cells; a method of treating a neurodegenerative disorder including administering a pharmaceutical composition comprising ventral midbrain-derived astrocytes and dopamine neural progenitor cells into a subject; and a method for differentiation into dopamine neurons, which includes preparing a mixture of ventral midbrain-derived astrocytes and dopamine neural progenitor cells and co-culturing or co-grafting the mixture. The co-grafting of ventral midbrain-derived astrocytes and dopamine neural progenitor cells (neural stem cells) improves the survival and differentiation of dopamine neurons and thus dramatically improves therapeutic outcomes in a neurodegenerative disorder including Parkinson's disease.

Disclosed is a method for producing an in vitro model for blood-brain barrier, including (a) a culturing conditionally immortalized astrocytes on one surface of a porous membrane and culturing conditionally immortalized brain pericytes on the other surface of the porous membrane, until both of the cells become a sheet; (b) culturing conditionally immortalized brain microvascular endothelial cells in a culture vessel, until the cells become a sheet; (c) peeling off the sheet of conditionally immortalized brain microvascular endothelial cells; (d) allowing the sheet of conditionally immortalized brain microvascular endothelial cells to come into contact with the sheet of conditionally immortalized brain pericytes, so that the sheets are arranged in layers; and (e) co-culturing a cell culture comprising three layers consisting of the sheet of conditionally immortalized brain microvascular endothelial cells, the sheet of conditionally immortalized brain pericytes, and the sheet of conditionally immortalized astrocytes.