19 results about "Cholangiocyte" patented technology

The invention relates to a composite hydrogel artificial bile duct as well as a preparation method and application thereof, the artificial bile duct is prepared by taking sodium alginate (SA) loaded with human umbilical cord mesenchymal stem cells and methylacryloylated gelatin (GelMA) composite hydrogel as raw materials, and the artificial bile duct can be used for biological materials or instruments for treating bile duct defects. The composite hydrogel artificial bile duct has better biocompatibility, can be tightly connected with an in-vivo tissue mucous membrane, and reduces stimulation to a local mucous membrane, so that bile duct obstruction caused by local tissue granulation hyperplasia is avoided, and adverse reactions are reduced; in addition, the composite hydrogel artificial bile duct can be automatically degraded after being implanted into a human body; the structure is compact, the mechanical property is good, and a good environment is provided for cell proliferation; the human umbilical cord mesenchymal stem cells can be directionally differentiated into bile duct cell-like cells, and bile duct injury repair is promoted.

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 present invention provides a method for generating hepatocyte and / or cholangiocyte lineage cells from pluripotent stem cells, the method comprising (a) specializing a prolonged nodal agonist-treated induced endoderm cell population by contacting the prolonged nodal agonist-treated induced endoderm cell population with a specialized medium to obtain a cell population including hepatic progenitor cells and / or cholangiocyte progenitor cells, the specialized medium comprising an FGF agonist and a BMP4 agonist and / or their active conjugates and / or fragments; (b) inducing maturation of the hepatic progenitor cells and / or cholangiocyte progenitor cells of the cell population and optionally further lineage specialization and / or expansion to obtain a population including hepatocyte lineage cells such as hepatoblasts, hepatocytes and / or cholangiocytes, the inducing maturation step comprising generating aggregates of the cell population. Optionally, the method also comprises activating the cAMP pathway within the aggregates and forming co-aggregates.

The present invention relates to an artificial bile duct comprising ursodeoxycholic acid (UDCA), and due to comprising UDCA, the artificial bile duct of the present invention is effective in improving differentiation into cholangiocytes and preventing formation of bile stones.

The invention belongs to the technical field of biological medicines, and particularly relates to an intestinal microbial marker for distinguishing a bile duct cell carcinoma patient from a healthy person and application of the intestinal microbial marker. The invention provides an enteric microbiological marker for distinguishing a bile duct cell carcinoma patient from a healthy person. The enteric microbiological marker consists of 23 enteric microbiological strains. The intestinal microbial marker disclosed by the invention realizes good distinguishing ability in cholangiocellular carcinoma and healthy people. And the feasibility and applicability of the distinguishing model are verified in cross-regional samples in China. In addition, the distinguishing model achieves good disease specificity in a common data set queue of 11 diseases from the world.

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.

Disclosed herein are improved pluripotent stem cell-derived biliary organoids, having extrahepatic cholangiocytes. In various embodiments, the biliary organoids can additionally include intrahepatic cholangiocytes. Also disclosed are methods of producing the biliary organoids having extrahepatic cholangiocytes. The disclosure also relates to methods of studying or treating a biliary-related disease or disorder using the described biliary organoids, including surgical methods such as in transplants, engraftments, ligations, and / or bililary bypass. The disclosure also includes surgical implants for biliary bypass. The disclosure additional describes methods of producing tubular organoids and cell culture devices for performing said methods.

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 invention belongs to the technical field of biological medicines, and particularly discloses an induced pluripotent stem cell (iPSC) strain carrying ABCB4 gene mutation and application thereof. The stem cell strain is preserved in the China Center for Type Culture Collection (the preservation number is CCTCC NO: C2025125), and carries ABCB4 gene composite heterozygous mutation c.992Ggt; a is (p.G331E), and c is 3152Tgt; the invention relates to the field of biomarkers (p.V1051A, C (p.V1051A), which can stably express pluripotent markers (OCT4, SOX2, NANOG and SSEA4), and has the capability of differentiating towards trigerm layers. The stem cell strain can be used for constructing a research model of progressive familial intrahepatic cholestasis type 3 (PFIC3), simulates disease phenotypes by differentiating hepatic cells or bile duct cells, is suitable for drug screening and gene therapy research, and provides a precise humanized tool for mechanism analysis and treatment development of ABCB4 mutation related diseases.

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 invention relates to the technical field of bile duct type organ culture, and particularly discloses a method for culturing a bile duct type organ based on postoperative drainage bile, which comprises the following steps: collecting bile, detecting cell survival rate, filtering, enriching stem cells by a multi-step centrifugation method, culturing the bile duct type organ and the like. The problems that direct surgical resection or biopsy specimen acquisition is limited, repeated sampling cannot be carried out for multiple times, and potential ethical disputes are avoided. By optimizing bile sampling and treatment processes, the method effectively solves the problem that excessive digestion or insufficient digestion is easy to occur when primary tissues are digested by traditional digestive enzymes, so that the purity and activity of bile duct cell separation are improved, the success rate of organoid culture is further improved, meanwhile, the operation steps are more concise and clear, and the method is suitable for large-scale popularization and application. And the technical requirements on operators are reduced, so that green hands can quickly grasp and successfully implement, and wide popularization and application of the technology are facilitated.

The present invention relates to methods for the expansion of cholangiocytes in vitro that comprise culturing the cholangiocytes in the presence of a farnesoid X receptor (FXR) agonist, such as chenodeoxylic acid (CDA) or obeticholic acid (OCA). The FXR-treated cholangiocytes and organoids obtained by the methods may be useful for example for the treatment of biliary disorders and compound screening. Also provided are kits and uses of culture media for the production of FXR-treated cholangiocyte organoids.

This invention relates to a method for the rapid and large-scale preparation of liver organoids, and to methods for screening drugs related to liver diseases and screening drug in vitro toxicity using these methods. According to the above-described preparation method, liver organoids can be rapidly and massively prepared by continuously undergoing a three-dimensional differentiation process on the same microplate. The massively prepared liver organoids using the above-described method include hepatocytes (solid core part), cholangiocytes (cystic part), hepatic stellate cells, and Kupffer cells, possessing a cellular composition, structure, and function similar to actual liver tissue, and can stably proliferate in vitro. Furthermore, the in vitro drug toxicity screening method for liver organoids of this invention allows for the pre-screening of drugs that may have hepatotoxicity, shortening the time and cost of new drug development and improving its efficiency.

The application provides a method for establishing a liver cancer immunotherapy efficacy prediction model based on the expression of paracancer bile duct cell chemokines and application thereof, and relates to the field of bioinformatics analysis. The application selects bile duct cells in liver cancer paracancer tissue as a key observation object, detects the expression intensity and range of three chemokines CXCL8, CCL2 and CCL20 in the bile duct cells by immunohistochemistry or immunofluorescence, quantifies the expression of CXCL8, CCL2 and CCL20 by using a semi-quantitative index such as H-score, quantitatively evaluates the expression of CXCL8, CCL2 and CCL20, and establishes a comprehensive scoring model by statistical analysis, which is used for predicting the efficacy benefit of a liver cancer patient receiving postoperative adjuvant immunotherapy, so as to make up for the deficiency of the lack of such specific molecular prediction method in the prior art, and provide a reliable basis for postoperative individualized immunotherapy decision-making.

This invention relates to the expansion of primary cholangiocytes in the form of cholangiocyte organoids (COs) using culture conditions in which canonical Wnt signalling is inhibited and non-canonical Wnt / PCP signalling is potentiated. Methods of expanding primary cholangiocytes, expanded populations of cholangiocytes and medical applications of expanded cholangiocytes are provided.

The present invention pertains to hepatocytes, liver progenitor cells, cholangiocytes, liver sinusoidal endothelial progenitor cells, liver sinusoidal endothelial cells, hepatic stellate progenitor cells, hepatic stellate cells, and liver cellular tissue models, as well as to methods for preparing these cells. The present invention also pertains to a cell fraction comprising liver progenitor cells, liver sinusoidal endothelial progenitor cells, or hepatic stellate progenitor cells. The present invention also pertains to a pharmaceutical composition or kit comprising the above-mentioned cells, a liver cellular tissue model, or a cell fraction. The present invention also pertains to: a method for screening liver disease treatment agents; a method for evaluating the hepatotoxicity of drugs, hepatocytes for infectious disease models, and a method for preparing the same; infectious disease model tissues and a method for preparing the same; as well as a method for screening infectious liver disease treatment agents.

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.