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Liver parenchyma cells and methods for their preparation, identification and application

A technology of hepatic parenchymal cells and hepatocytes, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve problems such as low efficiency

Active Publication Date: 2018-05-18
PEKING UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in the liver transplantation experiment of differentiated cells, the integration of individual human cells can only be detected in individual mice, and the efficiency is very low

Method used

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  • Liver parenchyma cells and methods for their preparation, identification and application
  • Liver parenchyma cells and methods for their preparation, identification and application
  • Liver parenchyma cells and methods for their preparation, identification and application

Examples

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Embodiment Construction

[0075] In the following examples, unless otherwise specified, the methods used are conventional methods, and the reagents used can be obtained from commercial sources. Wherein, bovine serum albumin component V (U.S. Calbiochem Company, 126579), human activin-A (Activin A, U.S. Peprotech Company, 120-14E), insulin-transferrin-sodium selenite mixed supplement solution (U.S. Invitrogen Company, 51300-044), B27 supplement solution (Invitrogen Company of the United States, 12587-010), hepatocyte culture medium HCM (Lonza Company of the United States, CC-3198), human fibroblast growth factor-7 (FGF7, Peprotech Company of the United States) , 100-19), SB431542 (U.S. tocris company, 1614), human bone morphogenetic protein-2 (BMP2, U.S. Peprotech company, 120-02), human bone morphogenetic protein-4 (BMP4, U.S. R&D systems company, 314-BP), Human hepatocyte growth factor (HGF, U.S. Peprotech company, 100-39), human oncostatin-M (OSM, U.S. R&D systems company, 295-OM), dexamethasone (Dex...

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Abstract

The invention discloses a hepatic parenchymal cell and its preparation, identification and application methods. The invention provides a method for preparing liver parenchymal cells, comprising the following steps: 1) culturing pluripotent stem cells to obtain endoderm cells; 2) culturing the endoderm cells obtained in step 1) to obtain primitive intestinal tube cells; 3) culturing the endoderm cells obtained in step 1) to obtain primitive intestinal tube cells; 2) the original intestinal tube cells were cultured to obtain hepatoblasts; it is characterized in that: the cells obtained from step 2) are subcultured, and transferred to the culture medium containing fibroblast growth factor (FGF) and bone morphogenetic protein (BMP). Culture in hepatocyte induction medium I to obtain hepatoblasts; 4) culture the hepatoblasts obtained in step 3) to obtain mature hepatocytes. The invention also relates to hepatocytes prepared according to said method. The hepatic parenchymal cells prepared by the invention have higher metabolic activity than before, high preparation efficiency and wide application prospect.

Description

technical field [0001] The invention relates to hepatic parenchymal cells and methods for their preparation, identification and application. Background technique [0002] The liver plays an important role in the metabolic process in the human body, including glycogen synthesis, urea synthesis, regulation of fat metabolism, and drug detoxification, etc. Damage to liver cells including hepatitis, liver cancer, and cirrhosis will cause great harm to human life. To solve this problem, it is necessary to obtain a large number of liver cells in vitro and use these cells for transplantation. Among various cell sources, human pluripotent stem cells (PSCs), including human embryonic stem cells (embryonic stem cells, ESCs) and human induced pluripotent stem cells (induced pluripotent stem cells, iPSCs), have the most application potential. Human embryonic stem cells have the ability of unlimited proliferation and differentiation totipotency, and can differentiate into various cell t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071A61L27/38C12Q1/02C12Q1/6881C12Q1/04G01N33/68
Inventor 邓宏魁赵东昕陈松
Owner PEKING UNIV
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