200 results about "Human hepatocyte" patented technology

The present invention relates to the preparation of non-human animals having chimeric livers, whereby some or substantially all of the hepatocytes present are human hepatocytes. It is based, at least in part, on the discovery that rats, tolerized in utero against human hepatocytes, were found to serve as long-term hosts for human hepatocytes introduced post-natally, and the introduced hepatocytes maintained their differentiated phenotype, as evidenced by continued production of human albumin.

One of the major hurdles of cellular therapies for the treatment of liver failure is the low availability of functional human hepatocytes. Although embryonic stem (ES) cells represent a potential cell source for therapy, current methods for differentiation result in mixed cell populations or low yields of the cells of interest. The present invention provides for a rapid, direct differentiation method that yields a homogeneous population of endoderm-like cells with 95% purity. In one embodiment, mouse ES cells cultured on top of collagen-sandwiched hepatocytes differentiate and proliferate into a uniform and homogeneous cell population of endoderm-like cells. The endoderm-like cell population was positive for Foxa2, Sox17 and AFP, and could further differentiate into hepatocyte-like cells that demonstrate hepatic morphology, functionality, and gene and protein expression. Incorporating the hepatocyte-like cells into a bioartificial liver device to treat fulminant hepatic failure improved animal survival, thereby underscoring the therapeutic potential of these cells.

Disclosed is a method for expressing hepatitis C virus in an in vivo, animal model. Viable, hepatitis C virus-infected, human hepatocytes are transplanted into a liver parenchyma of a scid/scid mouse host. The scid/scid mouse host is then maintained in a viable state, for up to five days or greater, whereby viable, morphologically intact human hepatocytes persist in the donor tissue and hepatitis C virus is replicated in the persisting human hepatocytes.

The present invention features a non-human animal model that is susceptible to infection by human hepatotrophic pathogens, particularly human hepatitis C virus (HCV). The model is based on a non-human, immunocompromised transgenic animal having a human-mouse chimeric liver, where the transgene provides for expression of a urokinase-type plasminogen activator in the liver. The invention also features methods for identifying candidate therapeutic agents, e.g., agents having antiviral activity against HCV infection. The animals of the invention are also useful in assessing toxicity of various agents, as well as the activity of agents in decreasing blood lipids.

A rotating wall vessel is used as a culture vessel and bioreactor for the cultivation of hepatocytes in the form of spheroids to generate a culture with many properties of the intact liver. These properties include enzyme activity comparable to fresh cells and long-term maintenance of viability and cellular function for periods on the order of months. The cultures may be used to produce hepatocyte products, evaluate metabolism of an agent, propagate Hepatitis C virus and test agents as inhibitors of this virus. Thus, the culture system disclosed herein makes long term functional cultivation of human hepatocytes feasible.

Anti-TRAIL-R1 and R2 antibodies or functional fragments thereof, having at least one property selected from the following (a) to (c) of: (a) having activity to induce apoptosis in carcinoma cells expressing TRAIL-R1 and/or TRAIL-R2; (b) not having effect on normal human cells expressing TRAIL-R1 and/or TRAIL-R2; and (c) not inducing human hepatocyte toxicity, and an anti-TRAIL-R1 and R2 antibodies or functional fragments thereof, having the following properties: having activity to induce apoptosis in carcinoma cells independently of exogenous factors and as a monomer of an antibody.

The present invention provides modified methods of producing human hepatocyte-like cells which exhibit phenotypes more similar to those of human hepatocytes. The present invention also provides the human hepatocyte-like cells and uses thereof. The methods of the present invention comprises a prolonged incubation period and addition of dexamethasone to the culture medium. The methods of the inventors produce human hepatocyte-like cells whose morphology is more similar to that of human hepatocytes, compared with the conventional system. The cells produced have both morphological and functional features of primary culture cells from normal human liver, including cytochrome P450 (CYP), multi-drug resistance-associated protein (MRP), and multi-drug protein (MDR). The use of human hepatocyte-like cells of the present invention enables, without using any animal model, assessment of metabolism and hepatotoxicity of test compounds, which are drug candidates, and screening for therapeutic agents for hepatic diseases, inhibitors to hepatitis virus infection, and therapeutic agents for viral hepatitis.

The invention discloses a method for separating primary adult hepatocytes. The method comprises the following steps: (1) carrying out multi-point puncture on surface of isolated adult hepatic tissue through a needle syringe, and injecting a preperfusate, wherein connective tissues are removed from the isolated adult hepatic tissue; (2) carrying out the multi-point puncture on the surface of the isolated adult hepatic tissue through the needle syringe again, and injecting a IV collagenase solution; (3) separating the isolated adult hepatic tissue, followed by adding the IV collagenase solution and carrying out digesting through vibration at a temperature of 37 DEG C to obtain digest; (4) carrying out filtering for the digest, followed by centrifuging and collecting cell aggregate in the underlayer, then resuspending the adult hepatocyte aggregate through a hepatocyte wash buffer, followed by filtering and centrifuging, then abandoning supernatant and collecting the adult hepatocyte aggregate in the underlayer; (5) washing the adult hepatocyte aggregate in the underlayer from the step (4) through a serum-free DMEM medium to obtain the primary adult hepatocytes. The invention further discloses a disposable special sterile apparatus box for separating the primary adult hepatocytes. With the present invention, the disposable special sterile apparatus box is adopted, the primary adult hepatocytes are separated through the multi-point puncture on the surface of the tissue and the injection, such that operation is simplified, cost is reduced, and the method and the apparatus box are applicable for extracting the hepatocytes from small pieces of the irregular isolated adult hepatic tissues of recovery of liver resection.

The invention provides an inula wissmannian extract and preparation thereof and application in preparation of an antitumor drug. The extract is gemma alkane type sesquiterpene lactone inula wissmannian lactone methyl, ethyl, propyl, butyl and amyl or sheepear inula herb lactone. The extract has the following chemical structural formula and absolute configuration that: due to the toxicity killing function experiment of inula wissmannian lactone methyl, ethyl, propyl, butyl and amyl or sheepear inula herb lactone on human hepatoma cells HepG2, human prostatic cancer cells PC-3, human gastric carcinoma cells MGC-803, human leukemia cells K562, human nasopharynx cancer cells KB and normal human hepatocytes LO2, the result shows that the gemma alkane type sesquiterpene lactone has strong tumor cell killing function, and proliferation of tumor cells can be inhibited, so that the tumor cells are killed. The animal in-vivo experiment proves that the inula wissmannian plant extract and monomeric compounds have the anti-tumor effect and can serve as active ingredients for preparing the antitumor drug. A novel cancer treatment medicine is provided clinically, and high clinical application values and social benefits are generated.

Disclosed is a mouse having human hepatocytes transplanted therein. Specifically disclosed is a mouse having human hepatocytes transplanted therein.  In the mouse, a foreign thymidine kinase gene or an urokinase-type plasminogen activator gene is retained so that the gene can be expressed specifically in the liver of the mouse, and hepatocytes of the mouse are substituted by human hepatocytes.

The present invention relates to biomedicine. Human hepatocyte growth factor full-length code area cDNA is inserted to defect duplicated adenovirus vector GT4050 to obtain recombinant adenovirus with carried human hepatocyte growth factor. When it is injected to rat body, the recombinant adenovirus has treating effect on rat's liver fibrillation. When it is applied to wound, the recombinant adenovirus can prevent scar.

The invention provides sgRNA and a vector pair for orientated knockout of an Nrf2 gene in human hepatocyte and application. The sgRNA consists of sgRNA1 for specifically recognizing the Nrf2 gene and sgRNA2 for specifically recognizing the Nrf2 gene; the vector pair consists of an Nrf2-CAS9-1 vector and an Nrf2-CAS9-2 vector; the Nrf2-CAS9-1 vector is a vector containing an sgNRA1 fragment; the Nrf2-CAS9-2 vector is a vector containing an sgRNA2 fragment; the vector pair can be applied to orientated knockout of the Nrf2 gene in the human hepatocyte and establishment of an Nrf2 gene knockout type human hepatocyte. By adopting the sgRNA, the vector pair and the application, the purpose of completely knocking out the Nrf2 gene at the genetic background that the length of a genome is not greatly changed can be achieved, and thus no Nrf2 protein can be retained in liver cells.

Niacin prevents and/or reverses the development of experimental hepatic steatosis, deposition of triglycerides in liver and serum of individuals, deposition of total cholesterol in liver and serum in individuals, and inhibit liver lipid peroxidation and oxidative stress in individuals at risk of developing fatty liver disease or NAFLD. In cultured human hepatocytes, niacin inhibits alcohol-induced fat accumulation.