25 results about "Neural crest" patented technology

Disclosed are in vitro methods for the differentiation of precursor cells into a neural crest cell (NCC) primed to a neurogenic lineage. The methods may include, for example, the steps of activating a Hedgehog signaling pathway (“HH signaling pathway”) in a precursor cell, wherein the precursor cell may be contacted with a neural crest cell induction medium for differentiation of the precursor cell into a neural crest cell. Compositions for carrying out the disclosed methods are also disclosed.

The present invention relates to a technology for developing a next-generation extracellular vesicle-based therapeutic agent and, more specifically, to a composition comprising three-dimensional stem cell-derived exosomes and neural crest cell-derived exosomes for alleviating neurological diseases. When extracellular vesicles of the present invention are administered in combination, progression of neuropathy can be effectively delayed or treated.

Provided are a preparation method for and a use of a dorsal root ganglion organoid. In one aspect, a preparation method for a dorsal root ganglion organoid is provided, which comprises: (1) inducing stem cells in a neural stem cell induction medium, to obtain neural stem cells; (2) inducing the neural stem cells in a neural crest cell induction medium, to obtain neural crest cells; (3) inducing the neural crest cells in a sensory neural progenitor cell induction medium, to obtain sensory neural progenitor cells; (4) inducing the sensory neural progenitor cells in an organoid induction medium, to obtain a dorsal root ganglion organoid. In addition, a use of the dorsal root ganglion organoid obtained by said method is also provided, for example, for drug screening or disease research.

The present invention provides a method for producing neural crest cells from pluripotent stem cells, the method comprising: a step (1) of subjecting pluripotent stem cells to suspension culture in a culture medium containing an ALK inhibitor and a bone morphogenetic protein; and (2) performing suspension culture in a culture medium containing an ALK inhibitor and a GSK-3beta inhibitor.

A method is provided herein for generating a human heart organoid (hHO) with integrated neural crest cells (NCCs), including incubating a neurosphere made from neural crest cells (NCCs) with an hHO, where the neurosphere is formed in a round-bottom or angled-bottom container. Additionally, the hHO has differentiated for about 11-17 days and / or expresses at least five times more Semaphorin-3C (SEMA3C) in comparison to an earlier hHO that has been differentiating for at least 48 hours less than the hHO. Also provided is an hHO with integrated NCCs prepared by the disclosed method. A method for evaluating the effects(s) of one or more drugs on hHO development and / or function by exposing NCCs to drugs, incorporating the NCCS into hHOs, and comparing drug-treated hHOs with integrated NCCs to non-drug-treated hHOs with NCCs is also provided.

The presently disclosed subject matter provides for in vitro methods of inducing differentiation of stem cells into hindbrain progenitor cells, enteric neural crest lineage cells, as well as differentiated hindbrain lineage or enteric neural crest lineage cells obtained by such methods. The presently disclosed subject matter also provides for uses of such cells for preventing and / or treating enteric nervous system disorders (e.g, Hirschsprung's disease), and for screening compounds suitable for preventing and / or treating enteric nervous system disorders (e.g., Hirschsprung's disease).

Human embryonic stem cell (hESC)- and induced pluripotent stem cell (iPSC)-derived clinical-grade corneal endothelial cells (CECs) generation and cryopreservation through the neural crest cell (NCC) lineage using peripheral blood as a donor source and hESCs maximizes the availability of these vital cells for treating the corneal endothelial disease as an alternative to donor corneas for corneal endothelium (CE) transplantations and other applications.

The invention relates to the technical field of biomedicine, in particular to a method for promoting formation of nerve fasciae and repair and regeneration after sciatic nerve defect by using small extracellular vesicles (hfNCSCs-sEVs) derived from hair follicle neural crest stem cells and application. According to the application disclosed by the invention, the effects of the hfNCSCs-sEVs on proliferation, migration, tube formation and barrier formation of nerve fascia cells in vitro are defined; the effect of hfNCSCs-sEVs on tissue regeneration and function recovery after rat sciatic nerve defect is defined, the hfNCSCs-sEVs inhibits expression of Smad7 in nerve tract membrane cells by transferring miR-21-5p, activates a TGF-beta / Smad signal channel, up-regulates expression of HAS2, promotes proliferation and migration of HAS2 and promotes expression of tight junction protein thereafter, and the hfNCSCs-sEVs can be used for treating rat sciatic nerve defect. And a new view angle and theoretical support are provided for repair and regeneration of sciatic nerve injury.

The invention discloses an induction method for directionally differentiating hESC-melanocytes based on a neural crest lineage, and belongs to the technical field of cell engineering and cytobiology. According to the induction method, a complex differentiation process of embryonic stem cells-ectoderm-neural ectoderm-neural crest cells is abandoned, and the neural crest cells are directly obtained from embryonic stem cells, so that the differentiation process is simplified; the differentiation efficiency and controllability of the neural crest cells are remarkably improved, the obtained melanocytes have higher tyrosinase activity and melanin synthesis capacity, and a new way is provided for treating leucoderma by adopting a melanocyte transplantation method.

This invention discloses a method for preparing and applying mesenchymal stem cells based on neural crest organoids, comprising the following steps: Step 1: Adding human induced pluripotent stem cells to a digestive solution, digesting, and then diluting with culture medium; Step 2: Collecting cells, centrifuging to separate the cells, discarding the supernatant, and suspending them in culture medium, adjusting the cell block size to 50-80 micrometers; Step 3: Culturing the cells, and then inducing differentiation into neural crest organoids; Step 4: Digesting the neural crest organoids into single cells, transferring them to serum-free mesenchymal stem cell culture medium for adherent culture, and then expanding to obtain mesenchymal stem cells; This invention utilizes induced pluripotent stem cells (ihPSCs)3D to induce differentiation into neural crest organoids, obtaining neural crest-derived mesenchymal stem cells, and the obtained NCC-MSCs have a beneficial and repairing effect on pancreatitis.

The present invention aims to provide a culturing method and a method for producing a cell population containing a neural crest cell, the methods being able to stably mass produce the neural crest cell from a pluripotent stem cell with quality that allows the cell to be used in a therapy with a cell, and a method for producing a mesenchymal stem cell and the like using the neural crest cell. The method for selectively culturing a neural crest cell in a cell population containing a neural crest cell and / or a neural crest progenitor cell of the present invention comprises a culturing step of adherent culturing the cell population containing the neural crest cell and / or the neural crest progenitor cell, in the presence of one or more extracellular matrices selected from the group consisting of laminin with its α chain being an α1 chain and its β chain being a β1 chain, laminin with its α chain being an α2 chain and its β chain being a β1 chain, laminin with its α chain being an α2 chain and its β chain being a β2 chain, and laminin with its α chain being an α5 chain and its β chain being a β1 chain, and extracellular matrices containing integrin-binding sites of these laminins.

The invention provides a parasympathetic neuron differentiation method which is characterized by comprising the following steps: (1) culturing iPSC (induced pluripotent stem cells) to form EB (Epstein-Barr) small bodies; (2) differentiating the EB small body into neurospinal cells; and (3) continuously culturing to obtain parsympathetic neurons. The parasympathetic nerve differentiation method is provided for the first time, and an effective tool and platform are provided for human parasympathetic nerve function and dysfunction research.

The invention relates to a preparation method and application of dorsal root ganglion organoid. On one hand, the invention provides the preparation method of the dorsal root ganglion organoid, which comprises the following steps: (1) inducing stem cells in a neural stem cell induction culture medium to obtain neural stem cells; (2) inducing the neural stem cells in a neural crest cell induction culture medium to obtain neural crest cells; (3) inducing the neural crest cells in a sensory nerve progenitor cell induction culture medium to obtain sensory nerve progenitor cells; and (4) inducing the sensory nerve progenitor cells in the organoid induction culture medium to obtain the dorsal root ganglion organoid. Moreover, the invention also provides application of the dorsal root ganglion organoid obtained by the method, for example, the dorsal root ganglion organoid is used for drug screening or disease research.

The present invention addresses the problem of providing a simple and efficient method for inducing differentiation of pluripotent stem cells into neural crest cells.　Provided is a method for producing neural crest cells, said method comprising a step for culturing stem cells for at least 6 days under conditions in which the expression or activity of a novel protein kinase C (nPKC) and / or an atypical protein kinase C (aPKC) is suppressed or inhibited.

The invention belongs to the technical field of biology, and discloses a construction method and application of a PAX3-labeled fluorescent report mouse model. According to the invention, a CRISPR-Cas9 gene editing technology is used, and an endogenous termination codon of a No.8 exon of a mouse PAX3 gene is accurately replaced with a P2A-EGFP sequence. According to the strategy, under the regulation and control of an endogenous PAX3 promoter and an enhancer, the co-expression of the fluorescent protein EGFP and the endogenous PAX3 protein is realized, and meanwhile, the integrity of the original structure and function of the PAX3 protein is ensured through a P2A self-cleavage peptide. The PAX3-P2A-EGFP report mouse constructed by the invention can specifically mark a plurality of early embryonic tissues such as neural crests, body segments, central nervous systems, muscle progenitor cells and the like in a high-fidelity manner, and an efficient and reliable in-vivo research tool is provided for dynamically tracking PAX3 related cell fate and researching related developmental diseases.

The present disclosure relates to a method of selectively differentiating and isolating, from pluripotent stem cells, neural crest cells having axial specification information in the body. According to the method of the present disclosure, initial neural crest cells, which correspond to an embryo neurulation stage and have not yet formed axial information in the body, can be produced, neural crest cells having specific axial information in the human body can be produced, and the cells can be specifically isolated and cultured in vitro such that various types of neural crest cells can be obtained all at once, and biological processes of neural crest cells for retaining axial information are researched to identify developmental processes of tissues and organs, and thus various cell therapeutic agents optimized for the characteristics of locations in the human body can be developed.

The present invention relates to a technology for developing a next-generation extracellular vesicle-based therapeutic agent, and relates to a composition for alleviating neurological diseases, comprising exosomes derived from neural crest cells. When the extracellular vesicles of the present invention are used, neuropathy can be effectively prevented or treated.

A method of producing a population of mature human pluripotent stem cell-derived sensory neurons (hPSC-SNs) expressing a target gene associated with at least one of nociceptive pain, chronic pain, pruriception and a nociceptive- or pruriceptive- mediated condition, the method comprising introducing into a population of human pluripotent stem cells (hPSCs) a composition comprising at least one site-directed nuclease targeting a site within the target gene, and at least one nucleic acid comprising a nucleotide sequence encoding at least one screenable, selectable marker that is flanked by (i) a nucleotide sequence homologous with a region located upstream of the target site within the target gene and (ii) a nucleotide sequence homologous with a region located downstream of the target site within the target gene, wherein the target site is located downstream of the open reading frame of the target gene, and wherein the site-directed nuclease cleaves the target site of the target gene and the nucleic acid encoding the screenable, selectable marker is inserted at the target site; covering said population of hPSCs under an extracellular matrix comprising at least one neuronal differentiation driver to produce a sensory committed neural crest population; contacting said sensory committed neural crest population with at least one neuronal differentiation driver and at least one neurotrophic factor to produce a population of early sensory neurons (SNs); and contacting said population of early SNs with at least one neurotrophic factor to produce a population of mature hPSC-SNs expressing at least one of SN marker or one pan neuronal marker; isolating the cells expressing the at least one screenable, selectable marker, wherein said population of mature hPSC-SNs expressed said one target gene associated with nociceptive pain, chronic pain, pruriception a nociceptive- or pruriceptive- mediated condition, and wherein said population of mature hPSC-SNs responds to a nociceptive and pruriceptive stimulus.

The present invention includes a functionalized nerve guidance conduit (NGC), methods of making neurotrophic factor-expressing neural crest stem-like cells (NCSC) and / or Schwann cell precursor-like (SCP) cells, methods of making the functionalized nerve guidance conduit, and methods of treating nerve injury using the functionalized nerve guidance conduit.

The present application specifically relates to a method for obtaining corneal endothelial cells, and the initial cells can be pluripotent stem cell-derived or neural crest cell-derived. Specifically, the present application discloses that pluripotent stem cells are efficiently induced to generate corneal endothelial cells by using a culture medium with completely clear chemical components. The present application also relates to CEC cells and their application in treating corneal endothelial diseases.

The invention discloses a method for constructing smooth muscle cells serving as a smoke disease model, and belongs to the technical field of crossing of stem cells and neuroscience. The method comprises the following steps: S1) reprogramming CD34 + cells in in-vitro PBMCs (peripheral blood mononuclear cells) of a smoke disease patient carrying RNF213p.R4810K mutation, so as to obtain induced pluripotent stem cells; s2) directionally inducing and differentiating the induced pluripotent stem cells into smooth muscle cells through a neural crest stem cell way, wherein the obtained smooth muscle cells are the smooth muscle cells capable of serving as the smoke disease model. The model can be used for researching the pathogenesis and nerve-blood vessel interaction process of smoke diseases, and can be further applied to molecular typing of diseases and in-vitro function evaluation of candidate drugs.

The invention discloses a method for detecting early neurodevelopment toxicity of pollutants based on a neural crest specific molecular marker and application, and belongs to the technical field of environmental toxicology and molecular diagnosis. The method comprises the following steps: 1) exposing an early embryo to a to-be-detected sample or pollutant; 2) detecting the change of the specific molecular marker of the neural crest and the peripheral neuron; and 3) evaluating the early neurodevelopmental toxicity according to the marker expression difference. According to the method, the problems of early warning lag and low sensitivity of the traditional neural behavior endpoint (zebra fish 6dpf) are solved, the early neurotoxicity of low-concentration pollutants can be detected within 24 hours of pollutant exposure (zebra fish sox10 and prph), and the method can be used for early toxicity early warning and risk assessment in environmental monitoring and chemical risk assessment.

The present invention relates to a method for selectively differentiating and isolating neural crest cells having human body axis information from pluripotent stem cells. According to the method of the present invention, it is possible to produce early neural crest cells that correspond to the embryonic neurulation stage and have not yet formed human body axis information, and to produce neural crest cells having specific human body axis information, which can be specifically isolated and cultured in vitro. This makes it possible to obtain various neural crest cells at once, and by studying the biological process of neural crest cells acquiring body axis information and elucidating the developmental mechanisms of tissues and organs, it is possible to develop a variety of cell therapeutic agents optimized for the location characteristics of the human body.

The invention provides a neural crest cell carrying a specific head-tail axis marker gene and a preparation method and application thereof, and relates to the technical field of biology, and the preparation method comprises the following steps: culturing stem cells to form an embryoid body; performing nerve induced differentiation on the embryoid body under the gradient driving of morphogenetic factors to form neural crest cells carrying specific head-tail axis marker genes, and promoting the formation of a neural tube-like structure with the polarity of the head-tail axis and the dorsal-abdominal axis when the embryoid body enters the nerve induced differentiation beginning by the gradient driving of morphogenetic factors. And inducing to generate neural crest cells carrying position-specific marker genes. The technical problem that neural crest cells carrying specific head-tail axis marker genes cannot be formed by an induced differentiation method in the prior art is solved.

The invention provides a craniofacial skeleton organoid controlled by sensory organs and sensory nerves and a preparation method thereof, and relates to the technical field of biologication.The preparation method comprises the steps that in a three-dimensional matrix, human pluripotent stem cells are induced through morphogenetic gradient driving to form nerve ectoderm with a nerve tube head-tail axis and a nerve tube back-abdominal axis; the neural ectoderm is respectively differentiated into a neural crest cell flow carrying head-tail axis position information, a visual cranium substrate-like structure carrying head-tail axis position information, an olfactory cranium substrate-like structure carrying head-tail axis position information and a trigeminal nerve cranium substrate-like structure carrying head-tail axis position information; and a craniofacial skeleton organ is formed under mediation of nerve cadherin. The technical problem that in the prior art, a method for forming a craniofacial skeleton organ through induction cannot form a sensory organ and a sensory nerve innervation function is solved.