The present invention discloses, in one embodiment, a method of using human induced pluripotent stem cells to generate three-dimensional human organ tissue for therapeutic drug toxicity and discovery⋅. In one embodiment, a high throughput microtiter plate is loaded with both wild type and Rett disease 3D spheroids and exposed to a drug library, and activity is measured and analyzed for disease rescue to wild type cell behavior.

The present invention provides differentiated neural cells and methods for making differentiated neural cells from pluripotent stem cells (PSC) at an industrial scale sufficient for high-throughput assays. The methods of the invention allow billions of PSCs and/or neural cells differentiated from the PSCs to be cryopreserved and expanded at multiple steps.

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 invention relates to modeling brain neuronal disease in a microfluidic device, comprising a co-culture of iPS-derived brain endothelial cells; iPS-derived dopaminergic neurons; primary microglia; and primary astrocytes, a Blood-Brain-Barrier (BBB)-Chip and a Brain-Chip. In particular, cross-talk between glial cells (e.g. microglia and astrocytes) with neuronal cells, in further contact with endothelial cells is contemplated for use for identifying drug targets under conditions for inducing in vivo relevant neuronal inflammation, neurodegeneration and neuronal death. Thus, in one embodiment, a microfluidic Brain-Chip comprising a co-culture of brain cells is exposed to α-synuclein preformed fibrils (PFF), a type of pathogenic form of α-synuclein. Such α-synuclein PFF exposure demonstrates an in vivo relevant disease pathogenesis on a microfluidic device as a concentration- and time-controlled manner that may be used for preclinical drug evaluation for diseases related to neuronal inflammation, e.g. Parkinson's disease (PD). In some embodiments, modulation of complement in the presence of neuronal inflammation is contemplated. In some embodiments, drug delivery to brain cells across the BBB is contemplated for preclinical testing of drug efficacy for slowing or stopping neuronal inflammation and degeneration.

Provided herein are, in various embodiments, methods and compositions for differentiating olfactory mucosa-derived mesenchymal stem cells (OM-MSC). In certain embodiments, the disclosure provides for media to differentiate OM-MSCs. In still further embodiments, the disclosure provides for methods and compositions using differentiated OM-MSCs for the treatment of nerve repair. In particular embodiments, the disclosure provides for novel treatments of peripheral nerve repair.

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.

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.

A cell-containing structure is provided that allows ready-to-use nerve drug response evaluation with high reproducibility to be easily performed. The cell-containing structure for evaluating an electrical property of neurons includes: (a) a culture surface to which the neurons are able to be adhered; (b) a cell mass that is adhered to the culture surface and contains at least one of the neurons; and (c) a plurality of electrodes for measuring the electrical property of the cell mass, wherein a spontaneous firing frequency of cells contained in the cell mass is 0.25 Hz or more per electrode.

Exemplary embodiments provide in vitro brain models, such as in vitro models of a neurovascular unit or a functionally connected trineural pathway, and systems, devices and methods of use thereof. The present invention provides in vitro brain models, systems, devices and methods that mimic in vivo conditions to, for example, determine the effect of a test compound, such as a drug candidate or a toxin, on various biological responses, such as for example, cell viability, cell growth, migration, differentiation and maintenance of cell phenotype, metabolic activity, structural remodeling and tissue level pre-stress, a neural activity, such as an electrophysiological activity.

Provided is a use of a Neurog2 functional fragment. The functional fragment can induce, in vivo or in vitro, glial cell formation into functional neuron cells, and thus can not only have a transdifferentiation function in normal tissue, but also facilitate neural reconstruction of damaged neural tissue.