76 results about "Neural crest" patented technology

The invention discloses an inducing method for directionally differentiating human embryonic stem cells to corneal endothelial cells. The method comprises the steps of: cultivating the human embryonic stem cells on a mouse embryonic fibroblast feed layer; sorting human embryonic stem cell clone groups in good state; grafting the groups on a human corneal stromal fibroblast layer processed by mitomycin C and cultivating for 7 days, wherein the human embryonic stem cells are differentiated to rosettes; separating and transferring the rosettes from the human corneal stromal fibroblast layer to a culture bottle; cultivating continuously for 7 days by using a neural crest stem cell culture medium; sorting the neural crest stem cells by a flow cytometry; adding the neural crest stem cells into the culture bottle; placing in a 5% CO2 incubator for incubating and cultivating at 37 DEG C by using a human corneal endothelial cell culture medium; changing the liquid every other day; and cultivating for about 10 days to obtain the corneal endothelial cells. The multiplication capacity of the corneal endothelial cells are similar to that of human corneal endothelial cells and the corneal endothelial cells can be transferred to 1-2 generations in vitro maximally. The corneal endothelial cells can be used as seed cells for cornea construction and transplant in tissue engineering.

A directional differentiation induction method of human embryonic stem cells to obtain corneal endothelial cells is provided. The method includes inducing the human embryonic stem cells to differentiate into human neural crest stem cells, inducing the human neural crest stem cells to differentiate into the human corneal endothelial cells, and other steps. The method adopts a primary human corneal stroma cell culture supernatant fluid, a human corneal endothelial cell culture supernatant fluid and a human lens cell culture supernatant fluid as a conditioned medium. Through adding retinoic acid and a plurality of recombinant proteins into the medium, and combining three-dimensional and two-dimensional culture methods, a corneal endothelial cell development process is simulated to induce the human embryonic stem cells to directionally differentiate into the human corneal endothelial cells, the corneal endothelial cells morphological structures of which are similar to those of normal human corneal endothelial cells can be obtained, and in-vitro subculture results show that proliferation of the obtained human corneal endothelial cells is consistent with proliferation of the normal human corneal endothelial cells.

The invention provides a method of efficiently producing corneal endothelial cells, particularly from corneal stroma or iPS cell-derived neural crest stem cells, a method of producing corneal endothelial cells stably in a large amount by inducing more efficient differentiation of stem cells into corneal endothelial cells, and a medicament containing corneal endothelial cells. The method of inducing differentiation of stem cells into corneal endothelial cells includes a step of culturing the stem cells in a differentiation induction medium containing a GSK3 inhibitor (preferably a GSK3β inhibitor) and retinoic acid, with the differentiation induction medium preferably further containing one or more of TGFb2, insulin, a ROCK inhibitor, and the like.

The present invention is based in part the discovery of methods for the generation of neural crest stem cells (NCSCs) from human pluripotent stem cells (hPSCs). Specifically, the present invention discloses methods for the use of a combination of rho-associated protein kinase (ROCK) inhibitors, glycogen synthase kinase 3 (GSK-3) inhibitors, activing receptor like kinase (ALK) receptor inhibitors and bone morphogenic protein (BMP) receptor inhibitors to derive NCSCs from hPSCs. The present invention also discloses methods to treat neurocristopathic diseases and disorders using NCSCs derived from hPSCs.

The present invention relates to compositions and methods employing postnatal (e.g., adult) neural crest stem cells. The stem cells are multipotent and differentiate when transplanted in vivo. Transplantation methods are provided for therapeutic, diagnostic, and research applications.

The present invention describes novel methods for isolating a substantially pure cell population of non-embryonic epidermal neural crest stem cells from the bulge-region of mammalian hair follicles. Also disclosed is the substantially pure cell population of follicular bulge-derived neural crest stem cells for medical research and therapeutic use.

The invention discloses a neural crest spectrum mesenchymal stem cell derived from pluripotent stem cells and an induced differentiation method thereof. The induced differentiation method of the neural crest spectrum mesenchymal stem cell comprises the following steps: dissociating the pluripotent stem cells to obtain cell aggregates, carrying out suspension culture to form embryoid bodies, replacing a culture solution with a neural crest cell culture solution, carrying out suspension culture of the embryoid bodies, collecting the embryoid bodies and replacing the culture solution every day; enabling the embryoid bodies to grow for a certain time by adhering to the wall after culturing for a certain time; digesting and collecting cells; culturing the collected cells by enabling the cells to adhere to the wall, replacing the culture solution with a mesenchymal stem cell culture solution, carrying out passage culture, then detecting the mesenchymal cell phenotype to obtain the neural crest spectrum mesenchymal cell. Through the method, the defects of heterogeneity and hybridity between adult-derived mesenchymal stem cells and other mesenchymal stem cells derived from the pluripotentstem cells of non-finite induction pathways can be solved; the prepared neural crest spectrum mesenchymal stem cell has higher immunoregulation and osteogenic differentiation capabilities; the standardized induced differentiation process is capable of ensuring that cell populations prepared between different batches have good consistency.

The present invention is directed to methods for treatment of melanoma using an inhibitor of dihydroorotate dehydrogenase (DHODH) and to combination therapies that involve administering to a subject an inhibitor of oncogenic BRAF (e.g. BRAF(V600E)), as well as an inhibitor of dihydroorotate dehydrogenase (DHODH). Assays for identifying compounds useful for the treatment of melanoma are also provided. The methods herein are directed to screening for compounds or agents that inhibit neural crest progenitor formation in a zebra fish model of melanoma.

The invention relates generally to polymorphisms or mutations of the PHOX2B gene. More particularly, the invention relates to polymorphisms or mutations of the PHOX2B gene that are responsible for the disease Hirschsprung's disease (HSCR), which is a neural crest-associated developmental disorder. Specifically, the invention relates to the detection of a single base-pair polymorphism in the PHOX2B gene that is associated with HSCR. The invention also relates to methods and kits for screening for carriers of mutations of the PHOX2B gene and the diagnosis of increased risk of HSCR. The invention further relates to diagnosing predisposition or susceptibility to increased risk of developing HSCR by screening for the presence of a polymorphism associated with HSCR. The invention also relates to compositions for screening for the polymorphism and treatment choices for patients having the polymorphism of the present invention. The invention further relates to providing polymorphisms in the PHOX2B gene for forensic use and in paternity test. The invention also relates to screening assays and therapeutic and prophylactic methods.

This invention relates in general to a mesenchymal stem cell (MSC) therapy. This invention further relates to the isolation and applications of gingiva derived mesenchymal stem cells. More particularly, this invention relates to the isolation and applications of the neural crest derived gingiva mesenchymal stem cells and/or mesoderm derived gingiva mesenchymal stem cells. This invention also relates to a composition comprising a neural crest derived gingiva mesenchymal stem cell and/or a mesoderm derived gingiva mesenchymal stem cell. This composition may be used for wound healing and/or in the treatment of inflammatory and/or autoimmune diseases.

The present invention relates to the differentiation of human pluripotent cells, including human pluripotent stems cells to produce a self-renewing multipotent neural crest cell population in a single step method without the requirement of isolation of intermediate cells and without appreciable contamination (in certain preferred instances, virtually none) with Pax6+ neural progenitor cells in the population of p75+ Hnk1+ Ap2+ multipotent neural crest-like cells. The multipotent neural crest cell population obtained can be clonally amplified and maintained for >25 passages (>100 days) while retaining the capacity to differentiate into peripheral neurons, smooth muscle cells and mesenchymal precursor cells.

The invention provides a novel method for isolating, culturing and differentiating neural crest stem cells from DRG (dorsal root ganglion). The method comprises steps as follows: (1) primary isolation of the neural crest stem cells from DRG; (2) purification and culture amplification of the neural crest stem cells from DRG; (3) induced differentiation of the neural crest stem cells from DRG. The obtained neural crest stem cells have higher multiplication capacity and still have multiplication capacity after being continuously cultured for generations in vitro.

Embodiments of the present invention are directed to methods for treatment of melanoma using an inhibitor of dihydroorotate dehydrogenase (DHODH) and to combination therapies that involve administering to a subject an inhibitor of oncogenic BRAF (e.g. BRAF(V600E)), as well as an inhibitor of dihydroorotate dehydrogenase (DHODH). Assays for identifying compounds useful for the treatment of melanoma are also provided. The methods comprise screening for compounds or agents that inhibit neural crest progenitor formation in a zebra fish model of melanoma.

This present invention features methods and composition for the isolation and proliferation of neural crest stem cells (NCSCs) from embryonic tissues as well as from tissues from a post-natal mammal. According to this invention, NCSCs are capable of producing non-neuronal and neuronal cells under the appropriate conditions. The cells of the invention therefore provide an accessible source for autologous and heterologous transplantation into the central nervous system, the peripheral nervous system, as well as other damaged tissues.

This invention is directed to an in vitro system for rapidly and uniformly inducing immortalized neural crest cells to differentiate to vascular smooth muscle cells. As excessive proliferation of vascular smooth muscle cells is a phenotypic response to the development of occlusive arteriosclerotic disease, the in vitro system of this invention is used to identify molecular regulators of smooth muscle cell development and differentiation. As the molecular regulators of smooth muscle cell differentiation are identified, the invention also encompasses methods to isolate the genes coding for these regulators. This invention also relates to molecules identified through the use of the invention's in vitro system, as well as to compounds that inhibit or regulate the identified molecules.

The invention discloses an ERK5 kinase and application of an ERK5 inhibitor in stem cell differentiation, and relates to the technology of stem cell differentiation regulation. Research shows that ERK5 plays an important role in the differentiation of stem cells and the ERK5 inhibitor can regulate the differentiation direction of stem cells; therefore, the ERK5 inhibitor has the application in thepreparation of any of the following reagents: (1) a reagent for regulating the differentiation of stem cells toward nerve cells, and the nerve cells are neural progenitor cells or neural crest cells;(2) an agent for regulating the differentiation of stem cells toward a germ layer, the germ layer is a trophoblast or mesoderm; and (3) an agent for promoting the differentiation of stem cells into cardiomyocytes. A new idea is provided for the differentiation of the stem cells.

Methods for enhancing beta cell survival and/or for stimulating beta cell proliferation, comprise co-transplanting pancreatic islets, beta cells, and/or stem cells which can generate beta cells, with (i) neural crest stem cells (NCSCs), (ii) tetracycline-regulated gene expression system (Tet-System)-containing neural stem/progenitor cells (TetStock neural stem/progenitor cells), and/or (iii) pre-differentiated stem/progenitor neural cells. Methods for reinnervation in an organ or tissue transplant patient comprise co-transplanting with the organ or tissue (i) neural crest stem cells (NCSCs), (ii) tetracycline-regulated gene expression system (Tet-System)-containing neural stem/progenitor cells (TetStock neural stem/progenitor cells), and/or (iii) pre-differentiated stem/progenitor neural cells. Kits for conducting such methods employ at least one of (i) neural crest stem cells (NCSCs), (ii) tetracycline-regulated gene expression system (Tet-System)-containing neural stem/progenitor cells (TetStock neural stem/progenitor cells), and (iii) pre-differentiated stem/progenitor neural cells.

The invention relates to the technical field of tissue engineering, in particular to a Schwarm progenitor cell derived from a marrow neural crest cell and application of the Schwarm progenitor cell to promotion of nerve regeneration. The rat marrow-derived neural crest cell N1, namely, BM-NCC clone N1, is provided, the preservation number is CCTCC NO:C201647, and the rat marrow-derived neural crest cell N1 has the performance of the Schwarm progenitor cell (SCP). The invention further provides a subculture method of the BM-NCC clone N1 and a method for induced directional differentiation to the Schwarm cell (SC). The BM-NCC clone N1 is expected to be applied to nerve regeneration promotion therapy based on cells.

The invention specifically discloses a neural crest stem cell preparation and application thereof. The neural crest stem cell preparation provided by the invention is obtained by culturing a neural tube in vitro, wherein the neural tube is derived from a 10-day pregnant mouse embryo. The neural crest stem cell preparation disclosed by the invention and obtained by culturing the neural tube derivedfrom a 10-day pregnant mouse embryo in vitro identifies neural crest stem cells by immunofluorescence; P0 generation cells and P3 generation cells respectively express characteristics of neural crestand do not express the characteristics of neural stem, which indicates that the cultured cells successfully maintain the characteristics of mouse neural crest stem cells in vitro; and in-vitro culture experiments show that the neural crest stem cells can grow stably in a provided culture medium and normal proliferation of cells is maintained.

The invention discloses a neural crest-lineage pericyte derived from multi-potential stem cells and an induced differentiation method of the neural crest-lineage pericyte. The induced differentiationmethod comprises the following steps: dissociating the multi-potential stem cells to obtain single-cell suspension, inoculating the single-cell suspension into a matrigel-coated pore plate, and changing the suspension through an NCN2 culture medium each day; carrying out culturing for a certain period of time, carrying out digestion, and collecting cells; carrying out adherent culture on the collected cells, changing culture liquid into pericyte induced culture liquid, carrying out subculture, and detecting the cell phenotype of the pericyte. According to the induced differentiation method, the disadvantages of limited source of human cerebral vessel pericytes, and the heterogeneity and linkage heterogeniety of the pericytes derived from multi-potential stem cells obtained in other non-finite induction ways are solved, and the obtained neural crest-lineage pericyte has very strong blood vessel stabilizing capacity and contractility; and by virtue of a standardized induced differentiation procedure, cell populations obtained in different batches have good coherence.