The present disclosure relates to a neural cell population, a neural cell-containing preparation, and a method for producing the population and preparation. More particularly, the present invention relates to a neural cell population derived from intraoral mesenchymal cells, wherein a proportion of normal diploid cells is 80% or more, a preparation containing the neural cell population, and a method for producing the population and the preparation.

Neurosupportive materials that possess strong tissue adhesion were synthesized by photocrosslinking two polymers, gelatin methacryloyl (GelMA) and methacryloyl-substituted tropoelastin (MeTro). The engineered materials exhibited tunable mechanical properties by varying the GelMA/MeTro ratio. In addition, GelMA/MeTro hydrogels exhibited 15-fold higher adhesive strength to nerve tissue ex vivo compared to traditionally used fibrin-based materials. Furthermore, the composites were shown to support Schwann cell (SC) viability and proliferation, as well as neurite extension and glial cell participation in vitro, which are essential cellular components for nerve regeneration. Finally, subcutaneously implanted GelMA/MeTro hydrogels exhibited slower degradation in vivo compared with pure GelMA, indicating its potential to support the growth of slowly regenerating nerves. Thus, GelMA/MeTro composites may be used as clinically relevant biomaterials to regenerate nerves and reduce the need for microsurgical suturing during nerve reconstruction.

The present invention provides a method for producing neural cells or a neural tissue, including the following steps (1)-(3):

(1) a first step of culturing pluripotent stem cells in the absence of feeder cells and in a medium containing 1) a TGFβ family signal transduction pathway inhibiting substance and/or a Sonic hedgehog signal transduction pathway activating substance, and 2) a factor for maintaining undifferentiated state,
(2) a second step of culturing the cells obtained in the first step in suspension to form a cell aggregate, and
(3) a third step of culturing the aggregate obtained in the second step in suspension in the presence or absence of a differentiation-inducing factor to obtain an aggregate containing neural cells or a neural tissue.

[Object]

Provided is a method of producing a nerve bundle including efficiently extending axons of neural cells.

[Solution]

Neural cells are cultivated in the presence of feeder cells including at least one type of cells selected from vascular component cells and perivascular cells.

The present invention relates to an axon bundle obtained by a method of culturing a nerve cell aggregation having one spheroid and an axon bundle extending from the spheroid wherein the nerve cell aggregation consisting of a plurality of neurons having a cell body and an axon obtained by a method comprising the following step: supplying a culture medium to one first chamber, one second chamber and one channel having the length of at least 1 mm, the width of 100 to 150 μm and the height of 100 to 200 μm, which connecting said first chamber and second chamber, wherein said first chamber, second chamber, and channel are contained in one of the modules disposed in a culture plate, seeding the first chamber with a nerve cell derived from a stem cell or a spheroid of primary cultured nerve cells; and culturing said nerve cell, thereby growing an axon bundle from said spheroid and extending them into said channel, wherein the length of the axon bundle obtained is 1 mm or more, and the diameter of the axon bundles is 50 μm or more.

A composition for regeneration of skeletal muscle includes a nerve cell secretome or an isolate thereof. An implantable therapeutic material for regeneration of skeletal muscle includes a hydrogel and nerve cells encapsulated within the hydrogel, wherein at least some of the nerve cells are living. A method of regenerating skeletal muscle includes applying the composition or implanting the implantable material adjacent to skeletal muscle myoblasts and/or myogenic cells to regenerate skeletal muscle therefrom.

A non-tubular material for nerve regeneration induction, which can be used for the regeneration of a damaged part in a nerve, and which comprises: (A) a crosslinked form produced by crosslinking a low-endotoxin bioabsorbable polysaccharide having a carboxyl group in the molecule with at least one crosslinkable reagent selected from a compound represented by general formula (I) and a salt thereof via covalent bonds; and (B) a bioabsorbable polymer. R.sup.1HN—(CH.sub.2).sub.n—NHR.sup.2 (I) [wherein R.sup.1 and R.sup.2 independently represent a hydrogen atom or a group represented by formula: —COCH(NH.sub.2)—(CH.sub.2).sub.4—NH.sub.2, and n represents an integer of 2 to 18]. Thus, a medical material that can induce the regeneration of a damaged part in a nerve is provided.

Provided is an isolated TrkB agonist antibody that binds to an epitope contained in one of the extracellular domains of TrkB and is capable of activating TrkB, wherein the extracellular domains comprises extracellular D1, D2, D3, D4, D5 domains and juxtamembrane domain of TrkB. Methods of using the TrkB agonist antibody in treating or reducing the risk of a TrkB associated conditions in a subject, wherein said condition is selected from cell differentiation, synaptic development, neural injury repairing and/or neurite branching.

A differential tissue-engineered nerve including motor-like nerves and sensory-like nerves. The motor-like nerve and the sensory-like nerve respectively includes a motor-like nerve outer tube and a motor-like nerve fiber in the outer tube as well as a sensory-like nerve outer tube and a sensory-like nerve fiber in the outer tube. Schwann cells and/or fibroblasts derived from motor nerves and sensory nerves are respectively contained in surfaces or pores of the motor-like and sensory-like nerve outer tubes. Transsynaptic signal molecules Neuroligin-1 and Neuroligin-2 are contained in surfaces or pores of the motor-like and sensory-like nerve fibers. Neuroligin-1 is selectively used to specifically promote synaptic remodeling of motor neurons, while Neuroligin-2 is selectively used to specifically promote synaptic remodeling of sensory neurons, so that repair efficiency of motor nerve cells and sensory nerve cells is improved.

It is a main object of the present invention to provide a process for producing a neuron-like cell from a somatic cell, a neuron-like cell obtained thereby, and a composition comprising a combination of chemical substances that can be used for said process without performing artificial gene transfer.

The invention can include, for example, a process for producing neuron-like cells, characterized by comprising a step of culturing somatic cells in the presence of two kinds of inhibitors, i.e., a TGF-β inhibitor and a BMP inhibitor, as well as any three or more selected from the group consisting of four kinds, i.e., a cAMP inducer, a GSK3 inhibitor, an Erk-inhibitor, and a p53-inhibitor, wherein the TGF-β inhibitor is a selective ALK5 inhibitor, and a neuron-like cell obtained by said process.

The neuron-like cells obtained according to the present invention are useful in regenerative medicine and the like.