42 results about "Neural crest" patented technology

The invention belongs to the field of drug preparation, and particularly relates to a preparation method and application of a drug delivery system based on hollow mesoporous polydopamine nanoparticles. The drug delivery system comprises hollow mesoporous polydopamine nanoparticles, a drug loaded inside and / or outside the hollow mesoporous polydopamine nanoparticles, and pectin wrapping the hollow mesoporous polydopamine nanoparticles, the medicine is a medicine with a nerve protection effect and an acetylcholin esterase inhibitor. The drug delivery system can reduce proinflammatory cytokines, regulate functions of target immune cells, synergistically reduce necrosis of neurons in intestinal tracts and induce neural crest stem cells in the intestinal tracts to be differentiated into neurons.

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.

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 invention discloses a preparation method and application of mesenchymal stem cells based on neural crest organics.The preparation method comprises the following steps that 1, human induced pluripotent stem cells are added into digestive juice, and after digestion, a culture medium is added for dilution; 2, collecting cells, centrifugally separating the cells, removing supernatant, suspending the cells in a culture medium, and adjusting the size of a cell block to 50-80 microns; 3, culturing the cells, and then carrying out induced differentiation on neural crest organs; 4, digesting the neural crest organs into single cells, transferring the single cells into a serum-free mesenchymal stem cell culture medium for adherent culture, and then performing amplification to obtain the mesenchymal stem cells. According to the invention, the induced pluripotent stem cells ihPSCs3D are induced and differentiated to obtain neural crest organs, the neural crest-derived mesenchymal stem cells are obtained, and the obtained NCC-MSCs have improvement and repair effects on pancreatitis.

The present invention relates to a method for selectively differentiating and separating neural crest cells containing human axial specialization information from pluripotent stem cells, and according to the method of the present invention, early neural crest cells corresponding to the embryonic neural tube formation stage and not having formed human axial information can be prepared. The neural crest cells with specific human body axial information can be prepared and subjected to specific separation and in-vitro culture, so that various neural crest cells can be obtained at a time, and the development process of tissues and organs can be revealed by researching the biological process of the neural crest cells with the axial information; thus, it is possible to develop various cell therapeutic agents optimized for body part characteristics.

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.

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.

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 invention belongs to the technical field of gene engineering, and particularly relates to a neural crest stem cell tracing system based on P75NTR-T2A-eGFP and application. The invention focuses on construction and application of a tracing system for differentiating pluripotent stem cells into neural crest stem cells in vitro. By constructing a Crispr / Cas9 vector, a termination codon region of a P75NTR gene expression cassette is precisely targeted. Meanwhile, a gene knock-in vector containing an (LHA) left homologous arm, T2A-eGFP-right homologous arm (RHA) is constructed. The two vectors are introduced into pluripotent stem cells (hPSCs) by means of electric shock transfection. Through the operation, the P75NTR-T2A-eGFP gene knock-in pluripotent stem cell line is successfully obtained, a foundation is laid for building a follow-up tracing system, and the P75NTR-T2A-eGFP gene knock-in pluripotent stem cell line has important application value in the field of neural crest stem cells.

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 relates to the technical field of cell biology, in particular to application of adult stem cells containing neural spine sources and a composition of the adult stem cells in treatment of pulmonary fibrosis. The composition has high loading capacity, pharmacodynamic experiments show that the composition can reduce the content of TGF-beta1, increase the percentage of SaO2 and reduce pCO2, can efficiently enter cells to play a role, inhibit and reverse pulmonary fibrosis caused by lung diseases and improve the pulmonary respiratory function, has a good treatment effect, is high in development value and has good application prospects. The method is suitable for further research and development and productization.

The presently disclosed subject matter provides for in vitro methods of inducing differentiation of stem cells into enteric neural crest lineage cells, and enteric neural crest lineage cells by such methods. The presently disclosed subject matter also provides for uses of such enteric neural crest lineage 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).

The present invention relates to a method for regulating the degree of differentiation of cells, comprising a step for forcibly expressing an MYC family gene and a BMI1 gene in a cell selected from the group consisting of (i) mesenchymal stem cells, vascular endothelial cells, smooth muscle cells, neural crest cells, and lymphocytes having an arbitrary degree of differentiation, and (ii) megakaryocytes and megayoung granulocytes having a high degree of differentiation.

The invention provides application of DNA tetrahedral framework nucleic acid in preparation of a medicine for preventing and / or treating fetal alcohol spectrum disorder, and belongs to the field of biological medicine. The DNA tetrahedral framework nucleic acid provided by the invention is formed by DNA single chains with sequences as shown in SEQ ID No.1-4 through base complementary pairing and self-assembly. The DNA tetrahedral framework nucleic acid can cross the placenta barrier, alleviate alcohol-induced neural crest cell apoptosis and oxidative stress injury, alleviate Et-OH-induced ROS level rise in vitro by regulating an Nrf2 / HO-1 pathway, and has a protection effect on fetal alcohol spectrum disorder craniofacial malformation and intrauterine growth retardation. The DNA tetrahedral framework nucleic acid provided by the invention has a good application prospect in preparation of drugs for preventing and / or treating fetal alcohol spectrum disorders.

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 invention belongs to the technical field of stem cell culture, and discloses a construction method and application for inducing and differentiating intestinal neural crest cells into colloidal precursor cells, the construction method comprises the following steps: S11, culturing the intestinal neural crest cells for 4 days to form neurospheres, sucking out supernatant containing the suspended neurospheres, centrifuging, discarding the supernatant, digesting with pancreatin, and collecting the supernatant; adding a DMEM / F12 culture medium to stop digestion, centrifuging, and discarding supernatant; s12, resuspending the neural spheres by using a colloidal precursor cell induction culture medium, inoculating the neural spheres into a cell culture bottle, and culturing in an incubator containing 5% of CO2 at 37 DEG C; and S13, after culturing for 16 hours, taking out the cells from the incubator to obtain the colloidal precursor cells. According to the method disclosed by the invention, intestinal neural crest cells are induced and differentiated into glial precursor cells and are further induced and differentiated into neurons, tool cells are provided for treatment of enteric nervous system diseases, a new tool is provided for understanding development and diseases of an enteric nervous system, and meanwhile, possibility is provided for developing a new treatment method.

To provide an alternative to donor corneas for corneal transplantation.SOLUTION: Human embryonic stem cell (hESC)- and induced pluripotent stem cell (iPSC)-derived clinical-grade corneal endothelial cell (CEC) 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) transplantation and other applications.SELECTED DRAWING: Figure 1

The present invention relates to a pharmaceutical composition for preventing or treating Alzheimer's disease, the composition comprising, as an active ingredient, neural crest-derived nasal turbinate stem cells (NTSCs) expressing SSEA3 and CD105. Treatment with the NTSCs expressing SSEA3 and CD105 or with an NTSC cell line including at least a predetermined proportion of the NTSCs was found to result in remarkably good therapeutic activity against Alzheimer's disease. Therefore, the present invention is expected to be effectively used not only as a composition for preventing or treating Alzheimer's disease in which the composition includes, as an active ingredient, NTSCs expressing SSEA3 and CD105 or an NTSC cell line including at least a predetermined proportion of the NTSCs, but also for uses such as screening of NTSC formulations that can be used to treat Alzheimer's disease, or prediction of the therapeutic efficacy thereof.

The present invention provides a method for producing corneal endothelial cells, the method comprising a step for culturing periocular neural crest cells by using a culture medium containing a fibroblast growth factor. The present invention makes it possible to provide corneal endothelial cells having characteristics close to that of corneal endothelial cells in a living body and having a quality enabling transplantation into a human.

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 provides a method for producing, from pluripotent stem cells, neural crest cells specialized for differentiation into a mesenchymal lineage, wherein the method includes: 1) a step for culturing pluripotent stem cells under xeno-free and feeder-free conditions in culture broth containing an ALK inhibitor and a GSK-3β inhibitor to obtain neural crest cells; and 2) a step for culturing the neural crest cells under xeno-free and feeder-free conditions in culture broth that includes an ALK inhibitor, EGF, and FGF and is substantially free of GSK-3β inhibitor.

The invention belongs to the technical field of genetic engineering, and particularly relates to a neural crest stem cell with high expression of KIF1BP as well as a preparation method and application of the neural crest stem cell. According to the invention, the KIF1BP is overexpressed in the NCSCs cells, so that the NCSCs KIF1BP can obtain more excellent migration ability and neuronal differentiation ability. According to the invention, high expression of KIF1BP is associated with migration of neural crest stem cells and neuronal differentiation capacity for the first time, and the limitation that the prior art depends on growth factors or physical induction to directionally differentiate NCSCs is broken through. Meanwhile, the autonomous migration ability of NCSCs is enhanced through high expression of KIF1BP, dependence on in-vitro pre-differentiation can be reduced, the colonization efficiency of transplanted cells in a focus area can be improved, and the treatment targeting and effectiveness can be remarkably enhanced when the neural crest stem cells are used for treating diseases, so that the effect of NCSCs in cell treatment is effectively improved.

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.