7 results about "Cholangiocyte" patented technology

The invention discloses a preparation method and application of exosome-loaded Cas13d-RNP, and belongs to the technical field of biliary atresia research. The method comprises the following steps: extracting exosomes: culturing human bile duct cells H69 in a DMEM / F12 culture medium until the cell density reaches 90%, collecting supernatant of the culture medium, and extracting the exosomes by an ultracentrifugation method; cas13d-RNP loading: the Cas13d protein and crRNA are mixed for 15 minutes at the room temperature, Cas13d-RNP is obtained, the exosome and the Cas13d-RNP are mixed, and the Cas13d-RNP is loaded into the exosome in a circulating freezing and thawing mode. According to the invention, Cas13d-RNP is accurately delivered through the exosome vector, specific targeting is carried out on bile duct cell CTGF mRNA, and expression of fibrosis-related factors in bile duct cells is significantly reduced. Compared with traditional medicine treatment, bile duct reaction and fibrosis processes can be more effectively inhibited. As a natural vector, the exosome has the advantages of low immunogenicity and high delivery efficiency, and immunoreactions possibly caused by a traditional virus vector are reduced.

The invention belongs to the technical field of biological medicines, and particularly relates to application of an MFN1 expression promoter in preparation of a medicine for treating mixed liver cancer. The MFN1 expression promoter provided by the invention is a recombinant expression vector containing MFN1. It is proved for the first time that MFN1 is highly expressed in hepatocellular carcinoma-like components of mixed liver cancer and is lowly expressed in cholangiocellular carcinoma-like components, and the MFN1 expression promoter can eliminate phenotypes of the mixed liver cancer by inhibiting cholangiocellular carcinoma-like tumor components in the mixed liver cancer, so that prognosis of tumor-bearing mice is improved. Theoretical basis and potential targets can be provided for treatment of the mixed type liver cancer, and certain guiding significance is achieved for research and development of targeted drugs for the mixed type liver cancer.

The present invention relates to a method for preparing a liver tissue-like organoid (LTO) from a stem cell-derived liver organoid, and relates to a method for preparing LTO which comprises culturing the liver organoid in medium M1 comprising human bFGF, human VEGF-A, human BMP4 and CHIR99021, and the LTO prepared by the method. The LTO of the present invention contains liver tissue-like biliary and vascular structures, as well as various hepatic cell types, including hepatocytes, cholangiocytes, hepatic stellate cells, endothelial cells, and immune cells, all of which maintain their specific functions. Therefore, the LTO can be usefully utilized in liver disease modeling, evaluation of toxicity and efficacy of drugs, and potential in vivo therapeutic applications.

The present invention relates of an ex vivo method for determining a reference proteomic profile of extrahepatic cholangiocarcinoma in the presence of bile duct stricture in a subject, comprising steps of : a) quantification of the protein abundances of all proteins identified in at least one bile duct cytological sample from at least one subject for which a diagnosis of extrahepatic cholangiocarcinoma has been previously established, or having benign stricture, b) quantification of the protein abundances of all proteins identified in human bile duct cells standard reference, c) determining the proteomic profile of each of said at least one sample by comparing protein abundances of each protein in the at least one sample with respect to the standard reference, d) determining a reference proteomic profile for extrahepatic cholangiocarcinoma condition, by means of a statistical test established with the proteomic profiles determined in step c). The present invention also relates to a reference proteomic profile of extrahepatic cholangiocarcinoma in the presence of bile duct stricture, and to an ex vivo method for differential diagnosis of extrahepatic cholangiocarcinoma in the presence of bile duct stricture of a subject.

The present disclosure provides a liver organoid model that recapitulates the hallmarks of non-alcoholic fatty liver disease (NAFLD), comprising both parenchymal and non-parenchymal liver cell types. The organoid is formed from primary hepatocytes isolated from healthy individuals, NAFLD patients, or a combination thereof, and expresses molecular markers of hepatocytes, cholangiocytes, stem cells, stellate cells, and Kupffer-like cells, displaying steatosis, inflammation, and fibrosis. The method for generating this model includes isolating primary hepatocytes, culturing them in defined initiation, expansion, and differentiation media, and performing functional characterization and passaging. The resulting organoid serves as a platform for developing in vitro models of steatohepatitis, high-throughput screening of candidate compounds, and gene expression and imaging analyses, with further validation in animal models. This approach enables physiologically relevant disease modeling and evaluation of therapeutic candidates for NAFLD and steatohepatitis.

The present invention relates to a method for preparing a liver tissue-like organoid (LTO) from a stem cell-derived liver organoid, and relates to a method for preparing LTO which comprises culturing the liver organoid in medium M1 comprising human bFGF, human VEGF-A, human PDGF-AB, human BMP4, human M-CSF and CHIR99021, and the liver tissue-like organoid (LTO) prepared by the method. The liver tissue-like organoid (LTO) of the present invention contains liver tissue-like biliary and vascular structures, as well as various hepatic cell types, including hepatocytes, cholangiocytes, hepatic stellate cells, endothelial cells, and immune cells, all of which maintain their specific functions. Therefore, the liver tissue-like organoid (LTO) can be usefully utilized in liver disease modeling, evaluation of toxicity and efficacy of drugs, and potential in vivo therapeutic applications.

The invention discloses application of HER-2 combined with a three-level lymph structure in diagnosis and treatment of mixed hepatocyte-bile duct cell carcinoma. A prognosis evaluation system and a treatment decision mode constructed on the basis of HER-2 detection are beneficial to establishment of a standard cHCC-CCA prognosis system, and the HER-2 detection can adopt a clinical conventional immunohistochemical method and is directly in butt joint with an existing pathological examination process, so that the clinical transformation threshold is effectively reduced, and the clinical treatment efficiency is improved. The clinical practicability and operability are obvious. Meanwhile, the expression level of HER-2 can reflect whether the patient can directly benefit from adjuvant chemotherapy, so that treatment schemes can be quickly generated for different patients after pathological examination. Moreover, the invention also discloses the association between the HER-2 and the tumor immune microenvironment, and provides a new thought for developing HER-2 and TLS combined prognosis products and targeted drug combined immunotherapy strategies.