Parenchymal hepatic cell and preparation, identification and application methods thereof

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: 2013-10-30
PEKING UNIV
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  • Summary
  • 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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  • Parenchymal hepatic cell and preparation, identification and application methods thereof
  • Parenchymal hepatic cell and preparation, identification and application methods thereof
  • Parenchymal hepatic cell and preparation, identification and application methods thereof

Examples

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Embodiment Construction

[0074] 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 parenchymal hepatic cell and preparation, identification and application methods thereof. The invention provides the method for preparing the parenchymal hepatic cell. The method comprises the following steps of: (1) culturing a pluripotent stem cell to obtain an endoderm cell; (2) culturing the endoderm cell obtained in the step (1) to obtain an original intestinal canal cell; (3) culturing the original intestinal canal cell obtained in the step (2) to obtain an adult hepatic cell; and (4) culturing the adult hepatic cell obtained in the step (3) to obtain a mature parenchymal hepatic cell, wherein the step (3) is characterized in that the cell obtained in the step (2) is subjected to passage and then transferred to a hepatic cell induction medium I containing fibroblast growth factors (FGF) and bone morphogenetic protein (BMP) for culturing to obtain the adult hepatic cell. The invention also relates to the parenchymal hepatic cell prepared by the method. The parenchymal hepatic cell prepared by the method has higher metabolic activity, high preparation efficiency and a wide application prospect than before.

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 ty...

Claims

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Application Information

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