13 results about "Hepatic stem cell" patented technology

The invention provides a method for detecting the influence of drug-containing serum of Sijunzi decoction on hepatic stem cells. The method comprises the steps of cell culture, drug-containing serum preparation, cell proliferation experiment, cell damage model establishment and treatment, oxidative stress related index detection, protein expression analysis and the like. According to the method, cell activity is detected through a CCK-8 method, cell nucleus morphological change is observed through Hoechst 33342 staining, the apoptosis rate is detected through Annexin V / PI double staining in combination with flow cytometry, apoptosis-related protein expression is detected through Western blot, and the action mechanism of the drug-containing serum is comprehensively evaluated. Experimental results show that the drug serum of the Sijunzi decoction can promote proliferation of liver stem cells, reduce oxidative stress injury induced by H2O2, improve cell viability and oxidation resistance, reduce cell apoptosis rate, up-regulate expression of anti-apoptotic protein Bcl-2 and inhibit expression of pro-apoptotic protein Bax and Cleaved Caspase-3 at the same time.

The present invention provides a method for producing hepatic stem cells, the method including a step for culturing hepatocytes in a medium containing a Wnt signaling promoting substance and an FGF signaling promoting substance. The present invention also provides a method for producing hepatic stem cells, the method including a step for culturing hepatic stem cells in a medium containing a Wnt signaling promoting substance, an FGF signaling promoting substance, and an ALK inhibitor.

The invention relates to the technical field of cell culture, in particular to nano liver stem cell peptide as well as a preparation method and application thereof. The preparation method of the nano liver stem cell peptide comprises the following steps: (1) mixing induced pluripotent stem cells with a first induced differentiation culture medium, and culturing for 3-5 days to obtain an intermediate cell 1; (2) inoculating the intermediate cell 1 into a second induced differentiation culture medium, and culturing for 3-5 days to obtain an intermediate cell 2; and (3) inoculating the intermediate cell 2 into a third induced differentiation culture medium, and culturing for 3-5 days to obtain the liver stem cell, and (4) after the liver stem cells are subjected to ultrafiltration, taking substances of which the molecular weight cutoff is less than or equal to 50kDa, centrifuging, taking precipitates, and drying to obtain the liver stem cell peptide. The liver stem cell peptide prepared by the method has the particle size of 1-20nm, can target the liver, has a good effect in treatment of liver injury, and does not generate drug resistance.

The invention discloses application of Rab8a in preparation of drugs for treating or preventing or improving individual hepatic fibrosis, and belongs to the technical field of drugs and disease treatment. The invention provides application of Rab8a in preparation of a medicine for treating or preventing or improving individual hepatic fibrosis. The Rab8a can enhance autophagy by down-regulating the expression level of the I-type TGF-beta receptor on the HSCs membrane, so that the activation of the HSCs is reduced, the hepatic fibrosis is favorably retarded or improved, and the Rab8a has the potential of serving as a target spot for treating the hepatic fibrosis.

The invention provides a support system for preserving an in-vitro liver through bidirectional driving of a liver and stem cell tank, which comprises an organ preserving device, a first circulating pipeline and a second circulating pipeline, and the first circulating pipeline and the second circulating pipeline are mutually independent; the organ storage device is used for storing the in-vitro liver; the first circulating pipeline is provided with a power part and a circulating purification unit, one end of the first circulating pipeline is connected with the inferior vena cava of the in-vitro liver in the organ storage device, and the other end of the first circulating pipeline is connected with the portal vein of the in-vitro liver; the second circulating pipeline is provided with a circulating pump and a bioreactor inoculated with mesenchymal stem cells, one end of the second circulating pipeline is connected with the inferior vena cava of the in-vitro liver in the organ preservation device, and the other end of the second circulating pipeline is connected with the hepatic artery of the in-vitro liver. The in-vitro liver is perfused through two channels, the purposes of detoxifying and supplementing essential factors can be achieved at the same time, and the in-vitro preservation time of the in-vitro liver can be effectively prolonged.

The invention relates to an exosome-containing composition, which is prepared by blending a modified silicon dioxide agent and an exosome matrix according to a weight ratio of (2-5): 15, the exosome matrix is composed of a mesenchymal stem cell source exosome and a liver stem cell source exosome, and the mass ratio of the mesenchymal stem cell source exosome to the liver stem cell source exosome is 1: (0.5-2). The exosome is a core functional component which plays a role in treating hepatic failure, the biological activity of the exosome is greatly improved through double-source selection and hypoxia pretreatment, the double-stem-cell exosome has a synergistic effect, proliferation and repair of damaged hepatic cells can be directly promoted, and liver functional cells can be supplemented; through compounding of the two exosomes, triple synergy of inhibiting necrosis, promoting regeneration and regulating immunity can be realized, and the problems of single function and limited repair efficiency of a single exosome are solved.

The present invention provides a method for establishing a hepatic stem cell line HepSC with bidirectional differentiation potential using pluripotent stem cells and its application. Specifically, the present invention provides a method for inducing human pluripotent stem cells to differentiate into HepSC, comprising the steps of: culturing pluripotent stem cells in a culture system to obtain functional HepSC, and continuing to culture HepSC to obtain functional hepatocytes or hepatocytes; and / or bile duct epithelial cells or bile duct epithelial cells. The functional HepSC, hepatocytes or hepatocytes; and / or bile duct epithelial cells or bile duct epithelial cells of the present invention have a very high differentiation rate and purity, and the HepSC, hepatocytes or hepatocytes; and / or bile duct epithelial cells or bile duct epithelial cells of the present invention can be used as an in vitro model for drug screening.

The invention relates to the technical field of cell culture, in particular to hepatic stem cells obtained by directional induced differentiation of mesenchymal stem cells as well as a preparation method and application of the hepatic stem cells. The induction method comprises the following steps: (1) inoculating mesenchymal stem cells into an induction culture medium, and culturing for 5-7 days to obtain intermediate cells 1; (2) inoculating the intermediate cell 1 into a differential medium, and culturing for 3-6 days to obtain an intermediate cell 2; and (3) inoculating the intermediate cell 2 into a mature culture medium, culturing for 3-6 days, filtering, centrifuging, collecting precipitate, and drying to obtain the liver stem cell. According to the method disclosed by the invention, the mesenchymal stem cells can be directionally converted into the liver stem cells. The liver stem cell obtained by the invention can be used for treating liver cirrhosis, has a good curative effect and no side effect, and provides a basis for treating liver cirrhosis.

There is disclosed a preparation of extracellular vesicles derived from human liver stem cells, preferably from a non-oval human progenitor cell line that expresses hepatic cell markers, which is capable of reducing the senescence of a population of senescent cells, as measured in a SA-β-galactosidase-based cellular senescence assay. Also disclosed are therapeutic applications of the preparation of extracellular vesicles derived from human liver stem cells according to the invention. Therapeutic applications include the reduction of cellular senescence, e.g. in an vitro or ex vivo method, as well as the therapeutic treatment of diseases and conditions known to be related to ageing and cellular senescence, such as for example atherosclerosis, diabetes mellitus type 2, asthenia, and others.

The present invention provides, inter alia: a method for inducing hepatic stem cells or hepatic progenitor cells from non-hepatic stem cells or non-hepatic progenitor cells; a method for producing hepatic stem cells or hepatic progenitor cells; and a method for producing stem cells or bile duct epithelial cells.

The invention discloses a method for three-dimensional multiplication culture of a hepatic stem cell line. The method comprises the following steps: inoculating cells of the liver stem cell line into a low-adsorption culture plate containing a HepSC culture medium, and culturing at the rotating speed of 100-180 rpm to enable the cells to form cell spheres. According to the invention, the dependence of the HepSC cell line on mouse matrigel and adherent culture is effectively solved, a new three-dimensional culture way without matrigel is provided for large-scale amplification, and clinical transformation of the liver stem cell line can be promoted.

The application discloses a method for constructing a tool mouse model for specifically expressing Cre in liver stem cells, and relates to the technical field of genetic engineering. The method comprises the following steps: after female mice are mated with male mice, fertilized eggs in a single cell stage are collected, and then a transgenic vector, sgRNA and Cas9 protein are microinjected into a male pronucleus; the fertilized eggs which develop into two cells are transplanted into the uterus of a pseudopregnant female mouse, and a transgenic F0 generation mouse is obtained through foster breeding; a transgenic founder mouse is screened out; the transgenic founder mouse is mated with a wild-type mouse, and a line is established through subculture to obtain an F1 generation mouse; the F1 generation mouse is subjected to genotype identification, and the tool mouse model is obtained. The tool mouse model constructed by the application can be applied to inducing LoxP recombination in liver stem cells, so as to conditionally knock out a target gene in liver stem cells, and the function of the target gene can be researched or a disease model can be constructed.

According to the present invention, methods are provided for inducing liver stem cells or liver progenitor cells from non-liver stem cells or non-liver progenitor cells, methods for manufacturing liver stem cells or liver progenitor cells, and methods for manufacturing hepatocytes or bile duct epithelial cells.