The present disclosure generally relates to a cell culturing system, and specifically to a three-dimensional cell culturing system for neuronal cells that promotes both structural and functional characteristics that mimic those of in vivo peripheral fibers, including cell myelination. Using a dual hydrogel construct and spheroids comprising neuronal cells, the present disclosure provides methods, devices, and systems for in vitro spatially-controlled, three-dimensional models that permit intra- and extra-cellular electro-physiological measurements and recordings. The three-dimensional hydrogel constructs allow for flexibility in incorporated cell types, geometric fabrication, and electrical manipulation, providing viable systems for culture, perturbation, and testing of biomimetic neural growth with physiologically-relevant results.

The present subject matter provides a microfluidic device that enables the precise and repeatable three dimensional and compartmentalized coculture of muscle cells and neuronal cells. Related apparatus, systems, techniques, and articles are also described.

The present invention is in the field of the cultivation of biological cells and tissues with organ-like function on a microphysiological scale and provides a method for the microphysiological co-cultivation of 3D organoid tissue and at least one 2D cell layer.

The invention is directed to a method for producing an edible composition, comprising incubating a three-dimensional porous scaffold and a plurality of cell types comprising: myoblasts or progenitor cells thereof, at least one type of extracellular (ECM)-secreting cell and endothelial cells or progenitor cells thereof, and inducing myoblasts differentiation into myotubes.

The invention relates to culturing motor neuron cells together with skeletal muscle cells in a fluidic device under conditions whereby the interaction of these cells mimic the structure and function of the neuromuscular junction (NMJ) providing a NMJ-on-chip. Good viability, formation of myo-fibers and function of skeletal muscle cells on fluidic chips allow for measurements of muscle cell contractions. Embodiments of motor neurons co-cultures with contractile myo-fibers are contemplated for use with modeling diseases affecting NMJ's, e.g. Amyotrophic lateral sclerosis (ALS).

The present disclosure describes the fusogenic promoting activity of the Myomixer protein. This polypeptide, when expressed in non-muscle cells with the Myomaker protein, is able to increase fusion of the cell with a muscle cell, but not with other non-muscle cells, as compared to cells only expression the Myomaker protein. The use of this protein and cells expressing it in the delivery of exogenous genetic material to muscle cells also is described.

An in vitro method of producing a population of muscle stem cells comprising co-culturing pluripotent stem cells, embryonic fibroblast cells and endothelial cells in 3D cell culture.

The present disclosure provides methods for generating myokines in vitro and compositions comprising myokines. Such compositions may be used for treatment of diseases such as cancer and fatty liver disease.

Isolated cells of a mixed character, possessing a mesenchymal stem cell phenotype and a muscle cell phenotype, as well as extracellular vesicles secreted from same, pharmaceutical compositions comprising same, and methods of treatment comprising administering same, are provided. Further, methods of increasing engraftment of foreign cells by co-administering same are provided.

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.