58 results about "Nerve stem cell" patented technology

The invention discloses a cultivated method of nervous stem cells, it includes following steps:1, Nervous stem cells substrate of human being is made up of base cultivate liquid which is compounded by mixing DMEM and F12 by rate of 1:1, and insulin, muriatic acid butyl amic, selenium natrium, ydrogen cortisone, L-glutamic acyl amic, man-turn iron albumen, flavone; 2, Noumenon serum of patient themselves is gathered immediately; 3. Noumenon serum of patient themselves is put into compounded Nervous stem cells substrate of human being, them, it is incubated by adding 3-7% CO2 warmer under conditino of 35C-38C. The method of the invention is simple, and invention has better repetition and its operation is simple and convenient; it plays an inestimable part in researching, teaching, clinic application, etc all of which relate to nervous stem cell and its application.

The invention provides icariin application in inducing stem cell in vitro orientation differentiates to several single type cells. The stem cell said includes embryonic stem cell, nerve stem cell and marrow mesenchyme stem cell. The single type cell includes nerve cell, bone cell, islet cells and endothelial cell. This invention also relates to application of the single type cell in preparing medicine of stem cell transplant curing nerve degenerative diseases, and its application in preparing cell differentiation agent that used to repair recovery injured nerve tissue, and its application in high efficiency drug effect screening model rebuilding and initial screening and estimating by using the model. The new applications of the icariin is extended in this invention, the fact of icariin contains pharmacy activity clarified, so it provides material basis for Chinese traditional medicine prevention and cure effect. The clarifying of drug effect mechanism provides reference for new medicine Chinese metical modern development with self-owned intellectual property right.

The invention relates to a nerve stem cell injection made from retina pigment epithelial cell, to treat paralysis agitans, wherein said injection at least comprises 3*106 nerve stem cells of human hRPE, while said nerve stem cells are cultivated from retina pigment epithelial cells. And its production comprises that: (1), separating and cultivating cells, that separating the hRPE cells, mixing uniformly to prepare single cell suspension; (2), hRPE cell cultivation and generation that digests original cultivation liquid, ending digestion, blowing and beating to form cell suspension, eccentrically filtering and removing the upper clear liquid, adding cultivating substrate, to disperse and form cell suspension, filtering with cell screen, transferring cell into cultivating bottle, generating to at least fifth generation; (3), adding common salt into hRPE nerve stem cell, to prepare cell suspension injection.

The invention discloses a method for preparing neural stem cells, which comprises a step of inductively culturing P19 cells in N2B27 culture medium to obtain a cell group containing neural stem cells above 94%. The method can culture the P19 cells in an environment without blood serum and retinoic acid, thus suppressing the interference of the blood serum with complex components and preventing the obtained neural stem cells from being transformed posteriorly by retinoic acid. Moreover, the method can directly transform pluripotent stem cells to neutral stem cells without resulting in selective cell apoptosis. The neutral stem cells obtained by the invention have anterior neutral plate characteristics and totipotency, and can well simulate the neurogenesis process in body. Accordingly, the neural stem cells can be used as the research model for analyzing neural induction and neural differentiation process from epiblast to neuroderm, thus providing an ideal path for researching development of embryo after nidation.

A nerve regeneration which comprises a compound is represented by formula (I):

(wherein all symbols are shown in the description), a salt thereof or a prodrug thereof. The compound of the present invention is suppresses nerve cell death as a substance for accelerating growth and/or differentiation of stem cells (nerve stem cells, embryonic stem cells, bone marrow cells, etc.), a substance for accelerating growth and/or differentiation of nerve precursor cells, a potentiator for neurotrophic factor activity, a neurotrophic factor-like substance or a neurodegenerative suppressor, and accelerates repair and regeneration of nerve tissues by neogenesis, regeneration and/or axon evolution. In addition, the compound of the present invention is useful for preparation from brain tissues, bone marrow and/or embryonic stem cells of cells for transplant (nerve stem cells, nerve precursor cells, nerve cells, etc.) and also accelerates grafting, growth, differentiation and/or function expression of cells for transplant whereupon it is useful for prevention and/or treatment of neurodegenerative diseases.

The invention discloses a bioreactor system of nerve stem cell's long-term expanding in big scale for bio-engineering technology field. This invention including: the air feed unit, the unit of nutrient fluid providing, the unit of cell expanding and increasing and as soon as wriggles pumping, its connection way is: The air feed unit terminal expands through the silica gel air conduit with the cell increases unit connected, the nutrient fluid supplies unit's terminal expands through the silica gel infusion tube with the cell increases unit connected, as well as wriggles pump in the silica gel infusion. This invention get out of the limit of raising box, and has advantageous for a long time greatly expanding and increasing scale and synchronous detect, and provides most suitable environment for nerve stem cell's raising in high density and long-term rasing.

The invention discloses a preparation method and application of a personalized induced neural stem cell exosome. The preparation method comprises the following steps: taking skin fibroblasts, carryingout subculture on the skin fibroblasts by using a liquid culture medium, and inducing the skin fibroblasts to become personalized induced neural stem cells by using a somatic cell reprogramming technology; and collecting a large number of exosomes secreted by the induced neural stem cells. The prepared personalized induced neural stem cell exosome has good skin penetrability and can directly permeate into the corium layer to improve the local regeneration microenvironment. Therefore, a skin anti-aging product prepared from the skin anti-aging composition has the effects of naturally keeping skin moist, whitening, resisting wrinkles and resisting pigmentation on normal skin, has the effects of promoting growth and recovering tissue integrity on aged and ultraviolet-damaged skin, has the effect of resisting scar formation on traumatic skin, does not irritate the skin, and is high in safety.

The invention belongs to biological medical and biological technology field. It relates to a vascular endothelia growth factor(VEGF) plasmid-liposome complex and its usage in the preparation of the pharmaceutical composition, in particular to the usage of VEGF plasmid-liposome complex in preparing pharmaceutical composition for treating cerebral ischemia injury, promoting differentiation of the adult cranial nerve stem cells and regenerating of the nerve cells. The complex of the VEGF and liposome is proved by the local cerebral ischemia animal model that said complex expresses highly of VEGF with only once injection, having no need of infusing for long period of time, having biological effects of brain protection and adult cerebral nerve stem cell differentiation promotion and regeneration of the nerve cell without the virus infection.

The invention provides a method for differentiation of amniotic fluid mesenchymal stem cells into neural stem cells, and the method comprises the following steps: centrifugally separating stem cells in amniotic fluid tissues to obtain the amniotic fluid mesenchymal stem cells; culturing the amniotic fluid mesenchymal stem cells by using an amniotic fluid mesenchymal stem cell serum-free proliferation culture medium; performing passage to P3-generation amniotic fluid mesenchymal stem cells; digesting the P3-generation amniotic fluid mesenchymal stem cells by using trypsin to obtain a single-cell suspension, and culturing the single-cell suspension overnight; removing the amniotic fluid mesenchymal stem cell serum-free proliferation culture medium, and then cleaning, and then adding the cells into a first induction culture medium for induction culture; and removing the first induction culture medium, then digesting the cells with trypsin to obtain a single-cell suspension, and carrying out inoculation culture on the single-cell suspension by using a second induction culture medium to obtain the cell ball formed by the neural stem cells. According to the invention, a variety of specific cell factors are used, and the amniotic fluid mesenchymal stem cells are induced by a two-step method to obtain the neural stem cells, wherein the obtained neural stem cells have the ball formationgrowth characteristics of neural stem cells.

The invention provides a neural stem cell specific gene delivery carrier and application thereof, and relates to the technical field of molecular pharmacology and molecular medicine. The delivery vector is a novel rAAV vector obtained by carrying out random gene mutation on the basis of a recombinant adeno-associated virus 9 and carrying out multiple rounds of in-vitro screening by taking neural stem cell spheres as a model. The amino acid sequence of the capsid protein of the delivery carrier is shown as SED ID NO: 1, the delivery carrier can efficiently deliver transgenic elements to neural stem cells in a targeted mode, compared with an rAAV9 carrier commonly used for transduction of a nervous system, the delivery carrier has the advantages that the transduction efficiency of the neural stem cells is remarkably improved, and the delivery carrier can be used for preparing drugs for diseases related to the nervous system.

The invention provides a medicine used for curing nerve damage and a method used for preparing the medicine. The invention belongs to the technical field of a biologic medicine. The medicine is prepared with snake venom nerve growth factors NGF, neurotrophy factors NTF, nerve stem cells and Schwann cells according to the following method: the entire spinal cord of a newborn animal or a fetus which can not be conceived because of natural and man-made calamities is implanted in aseptic PBS liquid or culture fluid; required ganglia are cleaned and shorn again and again. After the required ganglia are cultured in a thermostat of 37 DEG C for 48 hours, little culture fluid is added; the required ganglia are cultured for eight days before the required ganglia are frozen and deposited for passage. Before operation, the frozen and deposited cells are revived, and are implanted in the pia mater spinalis of the damaged spinal cord. The medicine can provide the cultured cells and the damaged neurocyte with new nutriments and NGF required for promoting the growth. The implanted cells can be separated and added to the damaged neural parts to repair the damaged, dead or feeble cells with safe and remarkable cure effect.

The invention discloses a channelrhodopsin-2 (ChR2)-green fluorescence protein (GFP) gene engineered nerve stem cell line and a construction method of the channelrhodopsin-2-green fluorescence protein (GFP) gene engineered nerve stem cell line. The cell line is the recombined nerve stem cell line C 17.2 of a channelrhodopsin-2 (ChR2) which is capable of stably expressing and a green fluorescence protein (GFP). The recombined ChR2-GFP gene engineered nerve stem cell line C 17.2 is obtained by constructing recombined slow virus, transducing C 17.2 nerve stem cell line and sifting. The channelrhodopsin-2 (ChR2)-green fluorescence protein (GFP) gene engineered nerve stem cell line can be used for in vivo and in vitro studies on the functional integration of a nerve neuron of a transplanted stem cell differentiation source and a nervous system of a host and provides a favorable platform for the study on the functional integration of a nerve neuron of a transplanted stem cell differentiation and a nervous system of a host. Especially with the utilization of a filter paper digestion method, the stem cell recombining method improves the rate of GFP positive cells, is beneficial to streaming Fluorescence Activated Cell Sorting (FACS) separation, and greatly improves the efficiency of an experiment.

The invention discloses a drug composition for treating cerebral injury and neurodegenerative symptoms in a recovery period and sequelae. Salvianolic acids and baicalin extracts are combined to be used in a compatibility manner, and effects of obviously improving local microenvironments and improving limb exercise and cognitive functions are realized. A pharmacology experiment proves that the composition can regulate and control expression of correlation factors of angiogenesis, and improves the local microenvironments. Multiplication of neural stem cells is promoted, the neural stem cells can be induced to be directionally differentiated towards a neuron direction, and functional nerve regeneration is promoted. The Composition containing effective ingredients of traditional Chinese medicines can be prepared into any drug preparations for curing the cerebral injury and the neurodegenerative symptoms.

The invention discloses application of components in promoting in-vitro proliferation of neural stem cells and further in neural stem cell transplantation. The Neural Stem Cells (NSCs) as the most promising cell type in stem cell treatment of neurological diseases have broad treatment prospects in treatment of Parkinson's disease, spinal cord injury, cerebral apoplexy and other diseases. ExogenousNSCs transplantation has high requirements for the number of neural stem cells. However, if only relying on NSCs conventional culture, the speed is limited. The invention finds that isonaringin has the activity of promoting in-vitro proliferation of NSCs, and the high concentration isonaringin has stronger promotion effect on the in-vitro proliferation activity of NSCs.

A method for co-culturing with nerve stem cells to inducing rASCs to be DA Neurons by Lmx1a mediation, belongs to a method for treating Parkinson's disease by inducing stem cells to replacing defective Neuron cells. The method of the invention comprises steps of: building Lmx1a gene clone, recombining plasmid, building, packaging and recombining adenovirus Ad-Lmx1a, and co-culturing rASCs and NSCs which are infected by Ad-Lmx1a to differentiating them to be DA Neurons; and culturing rASCs and NSCs passage cells. Protein marker for expressing dopaminergic neuron cells can be differentiated from inducted neuron-like cells in the invention, and the inducted cells can transmit electric signal or secrete neurotransmitters, to prove that the inducted cells are the Neuron cells.

The invention relates to a programmed cooling method for cells, in particular to a programmed cooling method for a human neural stem cell working cell bank. The programmed cooling method of the human neural stem cell working cell bank comprises the following steps of: (1) preparing a cryopreservation solution, namely adding a protective agent into a centrifuge tube, shaking a culture medium to achieve uniform mixing, adding a nutritional agent, and performing pre-cooling; (2) performing cryopreservation of neural stem cells,namely, collecting the cells in the centrifuge tube, performing counting, adding a pre-cooled cryopreservation solution, performing resuspending, sucking a cell suspension into a cryopreservation tube, and performing programmed cooling in a programmed cooling instrument; and (3) after finishing the programmed cooling, transferring the cell suspension into a liquid nitrogen tank for storing the suspension. According to the method of the invention, under the condition of programmed cooling, the proper cryopreservation solution is selected, the survival rate of cells under cryopreservation is not influenced when the temperature during cell resuscitation is within a relatively large range, the activity of the cells is higher after the cells are cryopreserved and resuscitated, and the survival rate of the cells is more than 95%.