The present disclosure provides a method of preparing mimicking angiogenic co-spheroids, including: co-cultring a neural related cell and a cultured cell on hyaluronan-grafted chitosan (CS-HA) substrates to form a co-spheroid of neural related cell/cultured cell, and encapsulating the co-spheroid of neural related cell/cultured cell into a hydrogel to form a mimicking angiogenic co-spheroid. The mimicking angiogenic co-spheroid of the present disclosure can be formed by 3D printing model as a 3D mini-neurovascular unit, which is applicated to a high-throughput angiogenesis screening platform.

Described here are systems and methods for deriving both spinal motor neurons and brain microvascular endothelial cells from induced pluripotent stem cells using distinct methods and combining them in a chip format. Neurons cultured alone in chip microvolume displayed increased calcium transient function and chip-specific gene expression. When seeded with endothelial cells, interaction further enhanced neural function, elicited vascular-neural interaction, niche gene expression with enhanced in vivo-like signatures arising from the chip co-cultures. Development of novel media formulations further allow for improved readout of differentiation process, by eliminating additives that otherwise confound differentiation processes and resulting phenotypes.

The invention provides a method of co-culturing mammalian muscle cells and mammalian motoneurons. The method comprises preparing one or more carriers coated with a covalently bonded monolayer of trimethoxysilylpropyl diethylenetriamine (DETA); suspending isolated fetal mammalian skeletal muscle cells in serum-free medium according to medium composition 1; suspending isolated fetal mammalian spinal motoneurons in serum-free medium according to medium composition 1; plating the suspended muscle cells onto the one or more carriers at a predetermined density and allowing the muscle cells to attach; plating the suspended motoneurons at a predetermined density onto the one or more carriers and allowing the motoneurons to attach; covering the one or more carriers with a mixture of medium composition 1 and medium composition 2; and incubating the carriers covered in the media mixture.

The invention relates to a cell sheet construct for neurovascular reconstruction. The cell sheet construct has a vascular endothelial cell layer and a neural stem cell layer, and the two layers are physically in direct contact with each other, where the vascular endothelial cell layer forms branching vasculatures, and the neural stem cell layer differentiates into neurons. The invention also relates to a method for manufacturing the cell sheet construct, having the following steps: culturing vascular endothelial cells on a substrate to form a vascular endothelial cell layer, seeding neural stem cells on the vascular endothelial cell layer to make the neural stem cells be physically in direct contact with the vascular endothelial cell layer, and culturing the neural stem cells and the vascular endothelial cell layer to differentiate into neurons and branching vasculatures to form a cell sheet construct.

Methods are disclosed for forming tissue engineered, tubular gut-sphincter complexes from intestinal circular smooth muscle cells, sphincteric smooth muscle cells and enteric neural progenitor cells. The intestinal smooth muscle cells and neural progenitor cells can be seeded on a mold with a surface texture that induces longitudinal alignment of the intestinal smooth muscle cells and co-cultured until an innervated aligned smooth muscle sheet is obtained. The innervated smooth muscle sheet can then be wrapped around a tubular scaffold to form an intestinal tissue construct. Additionally, the sphincteric smooth muscle cells and additional enteric neural progenitor cells can be mixed in a biocompatiable gel solution, and the gel and admixed cells applied to a mold having a central post such that the sphinteric smooth muscle and neural progenitor cells can be cultured to form an innervated sphincter construct around the mold post. This innervated sphincter construct can also be transferred to the tubular scaffold such that the intestinal tissue construct and sphincter construct contact each other, and the resulting combined sphincter and intestinal tissue constructs can be further cultured about the scaffold until a unified tubular gut-sphincter complex is obtained.

Human pluripotent stem cells are differentiated in vitro into forebrain subdomain structures, which are then fused to generate an integrated system for use in analysis, screening programs, and the like.

Disclosed herein are compositions of gastrointestinal organoids comprising cells originating from all three primary germ layers and methods of making and use thereof. These gastrointestinal organoids exhibit complex cellular organization and functions resembling naturally occurring organ tissue, and serve as excellent three dimensional models for studying gastrointestinal physiology.

Disclosed are methods that include separately differentiating stem cells to form cortical neural progenitor cells and differentiating stem cells to form dopaminergic neural progenitor cells. The cortical progenitor cells are transferred to a first compartment of a culture dish and the dopaminergic progenitor cells are transferred to a second compartment of the culture dish, separated by a removable barrier. The cortical progenitor cells and dopaminergic progenitor cells are cultured in medium that promotes differentiation of cortical neurons and dopaminergic neurons, respectively. The barrier is then removed, creating a single chamber containing the cortical progenitor cells and the dopaminergic progenitor cells, separated by a gap. The cortical progenitor cells and the dopaminergic progenitor cells are then cultured in a single medium, which promotes further differentiation of the neurons and also supports formation of connections (such as synapses or synapse-like structures) between at least some of the cortical neurons and dopaminergic neurons.

The present invention relates to substantially planar vascularized brain cancer organoid and methods of using such vascularized brain cancer organoids in anti-cancer drug discovery screen. In particular, provided herein are methods of producing and using complex, highly uniform vascularized brain cancer organoids that comprise physiologically relevant human cells and have the high degree of sample uniformity and reproducibility required for use in high-throughput screening applications.

A composition including adult pluripotent olfactory stem cells is provided. The adult pluripotent olfactory stem cells are obtained by culturing a cell mixture from an olfactory tissue of a mammal in media containing growth factors and then isolating cells which express B-lymphoma moloney murine leukemia virus insertion region-1 (Bmi-1).