42 results about "Liver Stem Cell" patented technology

The invention relates to the technical field of biomedical engineering and particularly relates to a long-term in-vitro culture and directional differentiation system and method for a liver stem cell. The long-term in-vitro culture and directional differentiation system comprises an amplification culture medium with specific chemical components, and a differential medium with specific chemical components, wherein the amplification culture medium is used for carrying out in-vitro culture of a mouse or human liver stem cell, and the differential medium is used for carrying out induced differentiation on the mouse or human liver stem cell to form a matured liver cell. By using the long-term in-vitro culture and directional differentiation system and method, a selectively-amplified liver stem cell in mother cells of the liver can be obtained from a mouse embryonic liver tissue or through human multipotential stem cell differentiation, the liver stem cell can be cultured for more than 20 generations under such a condition, and the stable molecular phenotype of the liver stem cell is maintained. By using the long-term in-vitro culture and directional differentiation system and method, the cultured mouse or human liver stem cell can be further subjected to induced differentiation to form the matured liver cell with functions of secreting albumin, metabolizing urea and the like.

The invention relates to a method for large scale preparation of liver stem cells, and uses of the liver stem cells in the fields of cell transplantation, stem cell culture, amplification, directional differentiation and tissue formation. The preparation method of the liver stem cells includes the following steps: extracting the liver stem cells from animal or human liver tissues through using a mechanical method, enzymatic method and mechanical-enzymatic method and density gradient centrifuge method combined technology; and carrying out in-vitro large scale culture, amplifying, carrying out liver stem cell identification, and freezing the liver stem cells for later use. The liver stem cell preparation efficiency of the method disclosed in the invention is about 100-10000 times higher than that of conventional methods, and the method can be used in tissue engineering and the production of liver stem cell preparations or stem cell drugs in order to provide a new method for the treatment of human diseases.

The invention provides a liver organoid model-based drug hepatotoxicity evaluation method. The evaluation method comprises the steps: 1, establishing a mouse liver organoid model: (1) taking mouse liver tissue, and sequentially carrying out cleaning, shearing, digestion, lysis and filtration treatment to obtain mouse liver stem cell precipitate; (2) preparing glue from the mouse liver stem cells,and performing culturing with a culture medium to obtain a liver organoid model; and 2, carrying out drug treatment on the liver organoid model: setting a drug to be detected into a plurality of concentration gradients, then giving drug stimulation under different concentration gradients to the liver organoid model, collecting organoid samples at different time points, observing morphological changes of the organoid samples, and detecting liver function indexes. The invention provides a more accurate method for in-vitro detection of drug hepatotoxicity, which has the advantages of short periodand simple operation, can replace the traditional cell line model and animal model, and is used for preclinical new drug development and clinical drug hepatotoxicity evaluation.

The invention relates to the field of stem cell culture, particularly a liver stem cell culture medium and culture method. The liver stem cell culture medium provided by the invention comprises a basal culture medium, HGF, SCF and LIF. The liver stem cell culture method provided by the invention comprises the following steps: separating the primary liver stem cells by immune magnetic beads, and culturing in the culture medium. The experiment indicates that the cell trauma is small in the primary cell obtaining process. The detection indicates that the activity is kept at 85% or so. The liver stem cells cultured by the method can enhance the propagation speed by 1.5-2.8 times. The flow cytometry detection indicates that the liver stem cells can keep favorable dryness.

The invention relates to a method for culturing liver stem cells. The method comprises the following steps of: a, flushing a liver tissue with PBS (Phosphate Buffered Saline) containing double antibodies for three times under the sterile condition; b, shearing the liver tissue, and digesting with a 0.25 percent g/mL trypsinase solution for 30 minutes; C, neutralizing digestive juice with a complete medium; d, beating and uniformly mixing the digested and neutralized liver tissue in a mixed liquid of the digestive juice and a neutralizing solution, centrifuging at the speed of 500 revolutions/minute for five minutes, extracting a supernatant liquid, and centrifuging the supernatant liquid at the speed of 1,500 revolutions/minute for five minutes; e, discarding the supernatant liquid, suspending precipitates with the complete medium, inoculating into a culture bottle, and culturing at the temperature of 37 DEG C in the environment of 5 percent CO2; f, culturing for four days, washing non-adherent cells, and changing the liquid with the complete medium; and g, changing the liquid with the complete medium once every four days, and performing passage in the ratio of 1:2 when the cells grow and expand to over 80 percent of the bottom area of the culture bottle.

A separation culture method for lens epithelium stem cells is disclosed. The method includes following steps: (1) cutting a lens anterior capsule into small pieces, digesting, and paving the obtained cells to a culture flask coated with a medium, a culture plate coated with a medium or a culture dish coated with a medium; and (2) adding an MEM culture solution containing 15-25% of FBS, 50-150 mug/L of a human epidermal growth factor (FGF), 5-10 mg/L of insulin, 5-10 mg/L of hydrocortisone, 5-10 mg/L of a vibrio cholera toxin, 0.01-0.05 mg/L of 3,3',5-iodo-L-para-hydroxyphenylalanine, 1-5 mg/L of adenine, 5-10 mg/L of glycine, 6-12 mg/L of alanine, 10-16 mg/L of asparagines, 10-16 mg/L of aspartic acid, 12-18 mg/L of glutamic acid, 8-15 mg/L of proline and 7-14 mg/L of serine, putting into an incubator having a temperature of 37 DEG C, culturing for 2-3 days to form clone, and forming lens stem cells with monolayer activity after 10-14 days. A separation culture solution for the lens epithelium stem cells is also disclosed. The method can separate and obtain the primary lens epithelium stem cells. The stem cells are high in activity. The method is simple, convenient, easy in operation and good in application prospect.

The invention provides a mouse liver organoid model, an establishing method therefor and application of the mouse liver organoid model. The establishing method comprises the following steps: withdrawing liver tissue of a mouse, and carrying out cleaning, shearing, digesting and filtering treatment sequentially, so as to obtain liver stem cells of the mouse; and subjecting the liver stem cells of the mouse to primary culture, propagation culture and differentiation culture sequentially, thereby obtaining the mouse liver organoid model. The liver organoid model obtained by employing the method is applied to pharmacologic, pharmacodynamic and toxicologic analysis. Compared with the conventional methods, the liver organoid model establishing method provided by the invention has the advantagesthat on one hand, the culture cycle is shortened greatly: the time cycle from material obtaining to model obtaining is about 14 to 20 days, the consumed time of the conventional culture method is 30 days, and thus, the feasibility of model application is improved obviously; and on the other hand, by the method provided by the invention, the success rate of mouse liver organoid culture is increasedobviously, and the growth speed of organoids of primary culture is accelerated.

The invention discloses a nutrition compound and application in preparing a drug for promoting stem cell proliferation. The nutrition compound is prepared from the following materials in proportion: lysine, methionine, phenylalanine, threonine, tryptophan, arginine, histidine, glycine, aspartic acid, leucine, isoleucine, valine, serine, glutamine, taurine, orotic acid, nucleotide, Vitamin A, Vitamin D, Vitamin B2, Vitamin B6, Vitamin B12, niacin, folic acid, Vitamin C, iron, zinc, manganese, copper, selenium, chromium, potassium, calcium, magnesium, inositol and granulesten. The nutrition compound has the effects of boosting the proliferation of hematopoietic stem cells, liver stem cells, mesenchymal stem cells, kidney stem cells and gastric mucosa stem cells. The nutrition compound can be used for preparing the drug for promoting stem cell proliferation, is free from toxic and side effect and can avoid various problems caused by introduction of exogenous stem cells for treating diseases.

The method of activating liver stem cell includes combined ethyl thioamino butyric acid and partial liver excision method, and conventional collagenase digesting and density gradient centrifugation method to separate oval liver cell. The method of the present invention can activate liver stem cell in short period without damage, and present invention makes it possible to provide great amount of cell for the preclinical research of liver stem cell and preclinical animal transplantation experiment.

The invention discloses application of nutrition combination in the preparation of a medicine for promoting the proliferation of liver stem cells. The nutrition combination can promote the proliferation of hemopoietic stem cells and the synthesis of hemoglobin, can provide conditions for awakening dormant adult liver stem cells and can further promote the proliferation of the adult liver stem cells. The invention solves all kinds of problems existing in the existing orthotopic liver transplantation and the donator liver stem cell transplantation, and has the advantages of high effectiveness, high safety, great convenience, economy, no immunological rejection and the like.

The invention relates to a liver stem cell injection and a preparation method. The injection is prepared by composing liver stem cells, umbilical cord blood stem cells, benzyl alcohol, asparagine, glutamine, isoleucine, proline, hyaluronic acid, transferrin, polygonatum polysaccharide, taurine, and progesterone according to a certain weight ratio. In the liver stem cell injection provided by the invention, the liver stem cells and umbilical cord blood stem cells are coordinated mutually to have better therapeutic effects on treating diabetes; the injection formula provided by the invention canmaintain the activity of liver stem cells for a long time, and the injection has no toxic and side components, is safe and effective, and low in cost, and can be directly applied to the clinical injection about diabetes.

The invention relates to the technical field of medical bioengineering, provides a liver stem cell specific surface molecular marker CD63 and a use thereof and especially relates to a use of CD63 in preparation of a kit for liver disease diagnosis or a drug for preventing or treating liver diseases. A method for fast separating, culturing and amplifying liver stem cells from liver by the molecular marker comprises fast separating liver stem cells from a liver cell suspension through the liver stem cell specific surface molecular marker CD63 through a flow cytometer. The liver stem cell with a CD63 positive result has the characteristics of self-renewal and bidirectional differentiation in vitro and has the effects of treating a damaged mouse liver. The separated liver stem cells can be used as seed cells for liver drug screening, tissular engineering liver and liver disease cell therapy and provide an ideal cell model for liver development and liver stem cell biological characteristic research.

This invention provides methods and systems for identifying and typing toxicity of chemical compositions, as well as for screening new compositions for toxicity. The invention involves detecting alterations in gene or protein expression and hence establishing molecular profiles in isolated mammalian LSCs contacted with various chemical compositions of known and unknown toxicities, and correlating the molecular profiles with toxicities of the chemical compositions.

The invention relates to a liver stem cell injection and a preparation method of the liver stem cell injection. The injection is prepared from liver stem cell, umbilical cord mesenchymal stem cell, polyethylene glycol, arginine, lysine, methionine, valine, glacial acetic acid, collagen, glycosaminoglycan, retinoic acid, and ethanol amine according to a certain component ratio. In the liver stem cell injection, the liver stem cell is matched with the umbilical cord mesenchymal stem cell, thus the treatment effect of diabetes is better; the injection formula can maintain the activity of the liver stem cell for a long time and is free from any toxic and side effects; the injection formula is safe and effective, low in cost, and can be directly applied to the clinical injection of diabetes. The liver stem cell cultivating method and its special culture base can effectively maintain the best state of the liver stem cell, guide the cell proliferation while keep the dryness of the liver stem cell, and reduce the damage rate and death rate of the liver stem cell in the cultivating and harvesting processes, and shorten the growth cycle.

An application of Bitaien in preparing the medicines in the form of capsule, oral liquid, particle, tablet, or injection for preventing and treating fatty liver, alcoholic liver, hepatitides, rheumatism, osteoporosis, gout, diabetes, cardiovascular and cerebrovascular diseases, etc, preventing cancer, delaying sanility, improving immunity, etc is disclosed.

The invention belongs to the field of biological medicine and particularly relates to a stem-cell biological preparation, and a preparation method and an application thereof. The stem-cell biological preparation comprises liver stem cells, peripheral blood stem cells, albumin and a solvent. The stem-cell biological preparation contains the liver stem cells, the peripheral blood stem cells and the albumin, and by the synergistic effect of the liver stem cells, the peripheral blood stem cells and the albumin, the tissue regeneration of the injured part of the liver can be effectively promoted, so that better liver-injury treatment effect is realized. Shown by experimental results, when the stem-cell biological preparation provided by the invention is adopted for treating a rat with acute liver injury, ALT, AST and TBIL indexes in the blood of the rat are respectively and obviously reduced, so that the liver injury of the rat can be obviously improved by adopting the biological preparation provided by the invention.

The invention discloses a method for combined induction of increase in endogenous Lgr5+ liver stem cells and an application of the method, and in particular a method for inducing the endogenous Lgr5+liver stem cells through combination of soluble protein molecules HGF and Rspo1. The soluble ligand can reach tissue stem cells through intravenous injection, and the soluble ligand, combined with a receptor, can activate the tissue stem cells. According to the method, the Lgr5+ liver stem cells are taken as targets. According to the method provided by the invention, on the basis of self-renewal and multifunctional differentiation capacities of various tissue and organ endogenous stem cells, the tissue and organ endogenous stem cells in a body are directly stimulated via the soluble molecules,so that self-renewal/regeneration of tissues and organs can be promoted; and from the aspects of application convenience, cost saving and safety, feasibility is enhanced.

The invention provides a culture medium for elutriating and amplifying liver stem cells and the application of the culture medium. Based on the clone formation capability of stem cells, chemical smallmolecules and cytokines in an elutriation culture medium are optimally combined, flow sorting of cell surface antigen markers is skipped, and a culture medium with clear components is researched anddeveloped. The optimized culture medium can be used for directly elutriating, separating and amplifying adult liver stem cells with liver/gallbladder bidirectional differentiation potential in adult livers, and has wide application prospect and huge market value.

The invention discloses a preparation and extraction method of liver stem cells. The method comprises the following steps: cleaning and disinfecting, washing the surface layer of liver tissue by usingclean water, scrubbing by using an alcohol pad, and then dipping in normal saline for 3min; performing enzymolysis, taking out the liver tissue, then washing clean by using a PBS buffer solution, cutting the liver tissue into small pieces to put in a protease solution, dissolving for 6-24h, and filtering an enzymolysis solution by using a filter screen; separating; screening, inoculating a basalculture medium with the separated enzymolysis solution until a cell cluster appears, observing cell shapes to determine the liver stem cells, and screening the liver stem cells; preserving. The invention relates to the technical field of bioengineering. By the preparation and extraction method of the liver stem cells, the screening process of the stem cells is simpler, the screening cost of the stem cells is lowered, meanwhile, impurities which are possible to appear in stem cell tissue fluid are effectively removed, and the research and use value of the extracted stem cells is improved.