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Preparation method for hepatic progenitor cells from different sources and application of hepatic progenitor cells

A technology of hepatocytes and hepatic parenchyma cells, applied in the field of cell models, can solve problems such as limited exploration of in-depth cytological characteristics and functions

Active Publication Date: 2020-03-24
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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Problems solved by technology

In the field of liver regeneration research, in recent years, many studies have traced different cell surface markers and found that when the liver is in a state of chronic injury, both parenchymal cells and non-parenchymal cells may transform into liver precursor cells. Cytometry uses fluorescent markers to separate hepatic precursor cells from different sources, and finds that they all have the potential of hepatic and biliary differentiation in vitro. In-depth cytological characterization and functional exploration of hepatic precursor cells from different sources is still quite limited

Method used

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  • Preparation method for hepatic progenitor cells from different sources and application of hepatic progenitor cells
  • Preparation method for hepatic progenitor cells from different sources and application of hepatic progenitor cells
  • Preparation method for hepatic progenitor cells from different sources and application of hepatic progenitor cells

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Embodiment 1

[0026] 1. Experimental materials

[0027] 1. Reagents related to cell culture

[0028] The recombinant protein HGF used in the present invention was purchased from Peprotech; and other small molecular compounds such as Y27632, A83-01 and CHIR99021 were purchased from TargetMol; the cells used were all derived from primary hepatocytes isolated from fresh liver tissue, and Use a specific cell culture medium (i.e., TEM proliferation medium) to culture and isolate type IV collagenase used in primary cells: purchased from Sigma; double combination antibiotics of penicillin and streptomycin purchased from Bo Rui Biopharmaceutical Company; William's E culture The base and HANKs solution were purchased from Shanghai Yuanpei Biological Company; the PBS buffer (Phosphate Buffer Solution) used for washing and resuspending cells was purchased from Gibco Company; the biological reagents used for frozen cells were purchased from Xinsaimei Company; The medium Dulbecco's modified Eaglemedium...

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Abstract

The invention relates to the technical field of cell models, in particular to a preparation method for hepatic progenitor cells from different sources. After induced culture is performed on hepatocytes and / or non-parenchymal cells with a hepatocyte transition and expansion medium, the induced-cultured hepatocytes and / or non-parenchymal cells transition into the hepatic progenitor cells which can expand and passage in vitro. According to the preparation method, by applying a lineage tracing method, the hepatocytes and the non-parenchymal cells in a mouse liver are isolated; by micromolecule assembly induced culture in vitro, the hepatocytes and the non-parenchymal cells successfully transition into the respective hepatic progenitor cells having an expansion and regeneration function; and meanwhile, proved by three-dimensional culture and organoid culture, the hepatocytes and the non-parenchymal cells both keep the characteristics of the cell sources thereof. The preparation method is conducive to further researching biological and morphologic characteristics of the hepatic progenitor cells from different sources, and further explores the application prospect of the hepatic progenitor cells as new transplanted cell sources.

Description

technical field [0001] The invention relates to the technical field of cell models, in particular to a preparation method and application of liver precursor-like cells from different sources. Background technique [0002] This year's study showed that the lineage tracing model marks cells with specific fluorescent signals to study their origin, evolution and final destination in different states of the liver (Tarlow, B.D., C. Pelz, W.E. Naugler, L. Wakefield, E.M. Wilson, M.J. Finegold, and M. Grompe, Bipotential adult liver progenitors arederived from chronically injured mature hepatitis. Cell Stem Cell, 2014.15(5):p.605-18.). Its advantage of full visualization makes it favored by various research teams. In the field of liver regeneration research, in recent years, many studies have traced different cell surface markers and found that when the liver is in a state of chronic injury, both parenchymal cells and non-parenchymal cells may transform into liver precursor cells. ...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0672C12N2500/30C12N2500/38C12N2501/11C12N2501/12C12N2501/415C12N2501/727C12N2509/00C12N2513/00
Inventor 鄢和新黄伟健王红阳
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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