39 results about "Human hepatocyte" patented technology

The present invention relates to methods for the in vitro production of hepatocellular organoids from primary human hepatocytes (PHHs). The invention also relates to a cell culture system for the in vitro expansion of functional fetal, neonatal, pediatric and adult PHHs and the mass production of organoids, and to the use of said cell culture system for the in vitro mass production and expansion of functional adult PHHs and / or the formation of hepatocellular organoids.

A method of inhibiting a cancer cell growth includes contacting the cancer cell with a Cu2(OH)3NO3 / CaSiO3@g-C3N4 nanocomposite material containing graphitic carbon nitride (g-C3N4), copper hydroxide nitrate (Cu2(OH)3NO3) and calcium silicate (CaSiO3), achieving an inhibition efficiency on human breast carcinoma (MCF-7) and human hepatocellular carcinoma (HepG-2) cell growth of greater than 95% in an in-vitro cellular viability assay.

The present disclosure provides an immunodeficient or immune-impaired chimeric non-human animal with a deletion or impairment of adeno-associated virus receptor (AAVR), comprising human hepatocytes, methods for preparing the chimeric non-human animal comprising human hepatocytes and methods of utilizing the chimeric non-human animal comprising human hepatocytes to evaluate transduction efficiency of adeno-associated viruses (AAV), and determine mechanism of inhibition / modification of AAV transduction in human hepatocytes.

This invention belongs to the field of biotechnology and pharmaceutical technology, and discloses an ACLY-targeting PROTAC chimera with anti-MASH activity using a glycol chain as the linking chain. Its general structural formula is Formula 1: R is -(CH2-O-CH2). n -, n is 2-6. The PROTAC compound synthesized in this invention is a novel compound. In a high-fat model established using L02 and HepG2 cells, the synthesized PROTAC compound exhibits activity in reducing TG content. Compared with the warhead, the synthesized PROTAC compound shows low toxicity activity in human hepatocellular carcinoma cells HepG2, normal human hepatocytes L02, and human umbilical vein endothelial cells HUVEC, making it a novel, low-toxicity, and highly effective drug for treating non-alcoholic steatohepatitis (NAH). This invention expands the application areas of PROTAC technology while developing a highly effective treatment for NHA.

A method of inhibiting cell growth in a cancer includes contacting a graphite-phase carbon nitride copper oxide and magnesium aluminum oxide (g-C3N4@CuO / MgAl2O4) nanocomposite with the cancer to inhibit growth of the cancer. The g-C3N4@CuO / MgAl2O4 nanocomposite comprises a graphite-phase carbon nitride (g-C3N4) in an amount of 2 to 18 percent by weight (wt. %), copper oxide in an amount of 1 to 10 wt. %, and magnesium aluminum oxide (MgAl2O4) in an amount of 75 to 95 wt. % based on a total weight of the g-C3N4@CuO / MgAl2O4 nanocomposite. The cancer includes cells from a cell line selected from a group consisting of a human hepatocellular carcinoma (HepG-2) cell line and a human breast carcinoma (MCF-7) cell line.

The invention discloses application of ginsenoside Rg1 in preparation of a product for treating oxaliplatin-induced liver injury, and belongs to the technical field of medicines. Experimental verification shows that after ginsenoside Rg1 is administrated, the survival rate of HL-7702 human hepatocytes induced by oxaliplatin is increased, the content of transaminase indexes AST and ALT in the cells is obviously reduced, the content of active oxygen is obviously reduced, and the condition of mouse liver injury can be obviously improved. The experimental results show that the ginsenoside Rg1 can improve the liver injury induced by oxaliplatin, and a new technical thought is provided for preparing the product for treating the liver injury induced by oxaliplatin.

The present application is used in the field of food and discloses a method for evaluating the degree of influence of trace components in wine on the level of cellular autophagy, comprising the following steps: Step 1, preparing a single cell suspension of human hepatocytes (LO2) stably expressing GFP-RFP-LC3 protein; Step 2, adding 1.5×10 4 The LO2 cells with a cell / mL were inoculated into a 6-well plate and covered with a glass for 24 hours until they adhered to the wall; Step three, 2 mL of wine containing 80 μL of pure alcohol or wine samples containing different trace components of wine were added to the cells and cultured at 37°C and 5% CO2 for 24 hours. Step four, fix in 4% paraformaldehyde for 15 minutes. Step five, wash the slides with PBS and dry them, then add an anti-fading agent containing DAPI, and place the side covered with cells on the anti-fading agent. Step six, observe the red and green fluorescence using a laser confocal microscope, and judge the effect of the trace components of the wine on the level of cellular autophagy based on the different fluorescence. The present application can quickly evaluate the degree of influence of the trace components in the wine on the level of cellular autophagy by observing the different fluorescence as mentioned above.

Disclosed herein are human hepatocyte culture media and methods that better support the fate restoration and maintenance of cultured human hepatocytes. Culturing human hepatocytes is improved with the supplementation of DMSO or DMSO2-supplemented hepatocyte clonal growth media or hepatocyte maintenance media with any one or a combination of the factors of fibronectin, laminin, vitronectin, serpin, and coagulation factors.

A method for evaluating the excretion of a substance of interest by a human hepatocyte-like cell comprises: providing a bladderlike-material-containing solution which contains a bladderlike material produced in vitro and having a membrane of a human hepatocyte-like cell and 0 to 10% by volume of a suspended extracellular matrix; adding the substance of interest to the bladderlike-material-containing solution to bring the substance of interest into contact with the bladderlike material; and removing the bladderlike material from the bladderlike-material-containing solution and measuring the concentration of the substance of interest or a metabolite thereof excreted into an inner cavity of the bladderlike material.

The present disclosure provides a chimeric non-human animal comprising human hepatocytes, methods for preparing the chimeric non-human animal comprising human hepatocytes and methods of utilizing the chimeric non-human animal comprising human hepatocytes to screening and identifying metabolites for any type of drugs, typically small molecule drugs, which might affect human liver functions and any other bodily function.

The invention provides a method for efficient site-directed gene knock-in in hepatocytes and application of the method. The invention provides a technical scheme for carrying out gene knock-in operation by taking proliferated human hepatocytes (Pro HHs) as target cells, which is characterized in that the gene knock-in operation is carried out by taking the proliferated human hepatocytes (Pro HHs) as the target cells. Through the optimization strategy of the system, the Pro HHs subjected to gene editing can be effectively recolonized and mature, and the treatment of diseases is successfully realized. According to the invention, powerful concept verification is provided for autologous gene edited proliferative cell treatment of human hereditary liver diseases, and a new way is opened up for treatment of liver diseases.

The invention discloses a benzopyran enantiomer and its preparation method and application. The present invention extracts and separates a pair of benzopyran enantiomers from the rhizome of Gentiana macrophylla, and evaluates the hepatoprotective activity of Gentiana macrophylla benzopyran by detecting the inhibitory effect of the benzopyran enantiomer on acetaminophen-induced human hepatocyte (L-O2) damage in vitro. It was found that the benzopyran enantiomer has good hepatoprotective activity. The preparation method of the benzopyran enantiomer provided by the present invention is simple to operate and low in cost, and the obtained benzopyran has high purity, a clear structure, and a variety of potential application values, which lays the foundation for the development of the medicinal value of Gentiana macrophylla benzopyran.

The present invention provides a product comprising plated human hepatocytes on a surface and at least some of the plated hepatocytes are in one or more hepatocyte clusters on feeder cells, which are attached to the surface. A method of preparing plated human hepatocytes is also provided. The preparation method comprises applying human hepatocytes to a surface in the presence of feeder cells, co-culturing the applied hepatocytes with the feeder cells, and forming one or more hepatocyte clusters by the co-cultured hepatocytes on the feeder cells, which are attached to the surface. The plated hepatocytes may be used for various purposes, including the preparation of a hepatitis B virus (HBV) infected hepatocyte culture model and drug testing.

The invention discloses an NK cell killing resistant cell strain as well as a construction method and application thereof. The NK cell killing resistant cell strain is named as a human liver cancer cell HepG2-NK-R Homo sapiens, and is preserved in the China Center for Type Culture Collection on November 05, 2025, and the preservation number is CCTCC (China Center for Type Culture Collection) NO: C2025298. The human hepatoma cell line HepG2 is exposed to NK cells for a long time for co-culture to obtain the human hepatoma cell line HepG2. The cell strain has a stable biological phenotype through in-vitro simulation of NK cell mediated long-term immune editing. Compared with parental cells, the cell line shows remarkably enhanced malignant biological behaviors, can be used for researching occurrence, development and metastasis of liver cancer or preparing a tumor cell model or a tumor animal model, is greatly improved in tumor forming ability, in-vivo proliferation speed and metastasis speed, can remarkably shorten an experimental period, and can highly restore invasion characteristics of tumors after evolution.

The invention belongs to the technical field of biomedicine, and relates to a culture medium for human placenta organoid in early pregnancy as well as preparation and application of the culture medium. The culture medium is prepared from DMEM / F12, and an N-2 additive is added into the culture medium; a B-27 additive; primary cell antibiotics; and N, N-acetyl-L-cysteine; l, L-glutamine; recombinant human epidermal growth factors; cHIR99021, CHIR99021; a recombinant human R-spinal protein 1; recombining a human fibroblast growth factor; recombinant human hepatocyte growth factors; a83-01, A83-01; prostaglandin E2; y-27632, Y-27632; and fetal bovine serum albumin. By improving the components of the culture medium of the human placenta organoid, the obtained human placenta organoid is better in growth state and balling rate, the apoptosis of the organoid is reduced, and a foundation is laid for further application to research of downstream molecular biology and cytobiology.

The invention discloses an application of isoalantolactone in preparation of a medicine for treating liver cancer, and through systematic in-vivo and in-vitro experimental research, the action effect of the isoalantolactone on normal liver cells of people and three liver cancer cell lines is evaluated. Various technical methods such as cell viability detection, phenotype analysis, marker protein detection, target verification, nude mouse ectopic transplantation tumor model and the like are adopted, and it is proved that the isoalantolactone plays a role in resisting liver cancer through specific targeting PCSK9. The method has the characteristics of simplicity and convenience in operation, good repeatability, remarkable effect and the like. The research discloses the application potential of the isoalantolactone as the PCSK9 targeted inhibitor in anti-liver cancer treatment for the first time, an important lead compound is provided for developing a novel PCSK9 targeted anti-liver cancer drug, and the discovery provides a new research direction and treatment strategy for targeted treatment of liver cancer.

The invention relates to the field of biological medicines, and discloses a composition and a medicine for relieving drug-induced liver injury as well as a preparation method and application of the composition and the medicine. The composition for relieving the drug-induced liver injury is prepared from clostridium praeparatum, mannan oligosaccharide, PQQ, IP6 and isogenistein. The inner layer structure of the medicine for relieving the drug-induced liver injury is used for embedding clostridium praeparatum and mannan oligosaccharide, and the outer layer structure of the medicine comprises PQQ, IP6 and isogenistein. The inner layer structure is formed by coating sodium alginate gel microspheres with chitosan; the outer layer structure is composed of hydroxypropyl methyl cellulose and sodium alginate. The survival rate of the composition for APAP-induced human hepatocyte injury reaches 85.5%, and hepatocyte injury and inflammatory response can be effectively relieved. The microcapsule structure enables the release rate of PQQ in gastric juice to be lower than 20%, the release rate of probiotics in gastric juice to be lower than 10%, and the total release rate of probiotics in intestinal segments to be higher than 90%, so that accurate delivery and stable protection of medicinal components are realized.

A method of treating a cancer by inducing apoptosis in a cancer cell includes contacting the cancer cell with a graphite-phase carbon nitride calcium metavanadate and calcium silicate (CaV2O6 / CaSiO3 / g-C3N4) nanocomposite. The cancer cell is selected from at least one cancer cell line from the group consisting of a human hepatocellular carcinoma cell line and a human breast carcinoma cell line.

Provided herein are completely defined culture conditions, including culture supplements and requirements for an extracellular matrix, for facilitating the expansion of and long-term maintenance of primary human hepatocytes, as well as culture conditions and cell culture media for maturation of hepatocytes.

The invention discloses an experimental method for verifying the effect of an anti-fatty liver compound, which comprises the following steps: S1, constructing a multi-level fatty liver model: respectively constructing a human hepatocyte line fatty degeneration model, an SPF-level mouse high fat diet induced fatty liver model and a db / db gene knockout mouse spontaneous fatty liver model, inducing fatty degeneration through a culture medium containing oleic acid and palmitic acid; s2, gradient treatment of the compound: configuring the anti-fatty liver compound to be verified into four concentration gradients of 0.1 mu mol / L, 1 mu mol / L, 10 mu mol / L and 50 mu mol / L; a multi-dimensional and multi-scale effect evaluation system is formed by integrating lipid metabolism related index detection at a cellular level, serum biochemical indexes at an animal level, liver tissue pathology and protein expression analysis and differential metabolite and pathway analysis at a metabonomics level, so that the improvement effect of a compound on the phenotype of the fatty liver can be intuitively reflected, and the effect of the compound on the phenotype of the fatty liver can be further improved. And the influence on a lipid metabolism molecular mechanism can be deeply revealed.

The invention relates to the use of a laminin (LN) matrix for hepatic differentiation. The invention also relates to a method for inducing hepatic differentiation comprising the steps of: (i) providing a population of human totipotent cells, (ii) culturing the population on a laminin-coated support in an endoderm stimulation medium to produce a population of human DE cells, (iii) culturing said population of human DE cells on a laminin-coated support in a hepatic stimulation medium to produce a population of human hepatoblast-like cells, and (iv) optionally culturing said population of human hepatoblast-like cells on a laminin-coated support in a hepatic maturation medium to produce a population of human hepatocyte-like cells.The invention further relates to a population of human hepatoblast-type cells or human embryonic hepatocyte-type cells obtained by the method of the invention. The invention further relates to a population of human hepatoblast-type cells expressing HNF4 α and expressing substantially AFP for use in a method of treating the human body.

The invention relates to a monoclonal antibody capable of recognizing a human Hepatocyte antigen, a preparation method of the monoclonal antibody and application of the monoclonal antibody in immunodetection. According to the technical scheme, 628-731 loci, 813-882 loci and 628-731 loci are selected to be connected in series to form the antigen peptide, codon optimization is carried out, the antigen peptide becomes a gene segment suitable for being expressed in escherichia coli BL21, and finally the obtained recombinant protein contains a Hepatocyte protein segment and a histidine protein tag. The recombinant protein is used for immunizing a mouse, and through cell fusion, screening and subcloning, a mouse hybridoma cell strain capable of secreting the anti-Hepatocyte protein monoclonal antibody and the anti-Hepatocyte protein monoclonal antibody secreted by the cell strain are obtained. The antibody obtained by the scheme has high specificity and sensitivity, can specifically recognize cells expressing Hepatocyte protein, and is suitable for immunological detection, especially immunohistochemical detection.

Provided are genetically modified non-human animals that are immunodeficient and optionally comprise xenotransplanted hepatocytes such as human hepatocytes, wherein the genetically modified non-human animal and / or the transplanted hepatocytes are modified to restore, activate, or increase interleukin-6 (IL-6) / interleukin-6 receptor (IL-6R) signaling pathway activity or interleukin-6 receptor subunit beta (GP130) signaling pathway activity and to restore, activate, or increase hepatocyte growth factor (HGF) / hepatocyte growth factor receptor (MET) signaling pathway activity in the transplanted hepatocytes. Also provided are methods of measuring or assessing the activity of human-liver-targeting reagents in such non-human animals and methods of making animals with a humanized liver (e.g., with reduced steatosis). Also provided are genetically modified, immunodeficient, non-human animals comprising a humanized HGF gene and optionally a humanized IL6 gene and methods of using and making such animals. Also provided are methods of increasing or accelerating engraftment of xenotransplanted hepatocytes in non-human animals.

The present invention provides a simple and robust human liver cell-based system in which persistent hepatitis C infection, persistent hepatitis B infection or ethanol exposure induces a clinical Prognostic Liver Signature (PLS) high-risk gene signature. The cellular model system for hepatocellular carcinoma (HCC) / cirrhosis development and progression may be used in the screening of compounds useful in the treatment and / or prevention of cirrhosis and / or HCC as well as in the identification biomarkers for the prediction of liver disease (especially cirrhosis) progression and HCC. The present invention also relates to specific compounds that have been identified, using such screening methods, as useful in the treatment and / or the prevention of HCC / cirrhosis.

The invention discloses an OX-Dex NPs preparation based on pH and spermine double cascade response, a preparation method and application, and belongs to the technical field of medicine preparation. The preparation method comprises the following steps: firstly, activating glucan by using excessive N, N '-carbonyldiimidazole, and then reacting with ethylenediamine to synthesize an amino-modified glucan intermediate; the preparation method comprises the following steps: firstly, preparing an amino-modified dextran intermediate, then connecting L-phenylalanine to the amino-modified dextran intermediate through a coupling reaction, and then self-assembling with oxaliplatin in the presence of cucurbit [8] uril and glutaraldehyde, so that the formed OX-Dex NPs preparation shows excellent cascade response characteristic to pH and spermine. Meanwhile, the OX (at) Dex NPs shows extremely low toxicity to human normal hepatocytes (LO-2) with low spermine expression and has remarkable toxicity to mouse breast cancer cells (4T1) with high spermine expression, and an efficient and safe technical scheme is provided for selection of breast cancer chemotherapy drugs.

The present invention provides an antibody that binds to an NTCP, capable of inhibiting infection of human hepatocytes with hepatitis B virus (HBV) particles. The present invention also provides an antibody that binds to an NTCP, capable of inhibiting infection of human hepatocytes with hepatitis B virus (HBV) particles, the antibody having a reduced effect on bile acid transport by NTCP.

The instant disclosure relates to genetically modified non-human animals comprising deletions of the Il2rg gene, the Fah gene and the Prkdc gene, as well as methods of making and methods of using the same.

The present invention relates to a nucleic acid construct comprising a UTR and use thereof. Specifically, the UTR can significantly improve the expression efficiency of a target gene in the nucleic acid construct. The nucleic acid construct may comprise a nucleic acid sequence expressing, for example, a human hepatocyte growth factor (hHGF), and can be used in a gene therapy for serious lower limb ischemia, diabetic foot, and other diseases.

This invention discloses the application of acyl-resorcinol derivatives in the preparation of drugs or health products for the prevention and treatment of non-alcoholic steatohepatitis (NASH). Studies have shown that in an in vitro model of free fatty acid-induced normal human hepatocytes, these compounds can exert a hepatoprotective effect by reducing lipid accumulation in cells. The representative compound, hyperacmotone A, significantly reduces lipid accumulation in cells in a dose-dependent manner and regulates the expression of genes such as Pparα, Cpt1, and Fapp1. In a methionine-choline deficiency-induced mouse model, administration of hyperacmotone A can inhibit hepatic steatosis, reduce serum alanine aminotransferase (ALT) and / or aspartate aminotransferase (AST) levels, while simultaneously reducing hepatic triglyceride levels and increasing hepatic glutathione activity. This suggests a certain preventive and therapeutic effect on the occurrence and development of NASH.

Provided herein are completely defined culture conditions, including culture supplements and requirements for an extracellular matrix, for facilitating the expansion of primary human hepatocytes.