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Preparation and extraction method of liver stem cells

A technology for liver stem cells and extraction methods, applied in cell dissociation methods, hepatocytes, biochemical equipment and methods, etc., can solve the problems that affect the research and use performance of stem cells, the complex stem cell process, and the high screening cost, so as to reduce the screening cost. , Improve the value of research and use, and facilitate the observation of the effect

Inactive Publication Date: 2018-11-16
安徽
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the deficiencies of the prior art, the present invention provides a method for the preparation and extraction of liver stem cells, which solves the problem that the existing stem cell extraction method has a complicated screening process for stem cells, high screening costs, and impurities that are easy to remain in the stem cell extraction process, which affects the research of stem cells. and performance issues

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] A method for preparing and extracting liver stem cells, comprising the following steps,

[0028] (1) Clean and disinfect, wash the surface of the liver tissue with clean water, wipe it with alcohol cotton, and then soak it in normal saline for 3 minutes;

[0029] (2) Enzymatic hydrolysis, take out the liver tissue and rinse it with PBS buffer, cut the liver tissue into small pieces and put it in the protease solution for 6 hours, and filter the enzymatic solution with a filter;

[0030] (3) Separation, pour 5 times of distilled water to dilute the enzymatic solution, stir and mix evenly with a clean glass rod, and pour into a differential centrifuge;

[0031] (4) Screening, inoculate the separated enzymatic solution into the basal medium until cell clusters appear, observe the shape of the cells to determine the liver stem cells, and screen out the liver stem cells;

[0032] (5) Preservation. Store the liver stem cells in a low-temperature room at a temperature of 4°C....

Embodiment 2

[0043] A method for preparing and extracting liver stem cells, comprising the following steps,

[0044] (1) Clean and disinfect, wash the surface of the liver tissue with clean water, wipe it with alcohol cotton, and then soak it in normal saline for 3 minutes;

[0045] (2) Enzymatic hydrolysis, take out the liver tissue and rinse it with PBS buffer, cut the liver tissue into small pieces and put it in the protease solution for 15 hours, and filter the enzymatic hydrolysis solution with a filter;

[0046] (3) Separation, pour 5 times of distilled water to dilute the enzymatic solution, stir and mix evenly with a clean glass rod, and pour into a differential centrifuge;

[0047] (4) Screening, inoculate the separated enzymatic solution into the basal medium until cell clusters appear, observe the shape of the cells to determine the liver stem cells, and screen out the liver stem cells;

[0048] (5) Preservation. Store the liver stem cells in a low-temperature room at a tempera...

Embodiment 3

[0058] A method for preparing and extracting liver stem cells, comprising the following steps,

[0059] (1) Clean and disinfect, wash the surface of the liver tissue with clean water, wipe it with alcohol cotton, and then soak it in normal saline for 3 minutes;

[0060] (2) Enzymatic hydrolysis, take out the liver tissue and rinse it with PBS buffer, cut the liver tissue into small pieces and put it in the protease solution for 24 hours, and filter the enzymatic solution with a filter;

[0061] (3) Separation, pour 5 times of distilled water to dilute the enzymatic solution, stir and mix evenly with a clean glass rod, and pour into a differential centrifuge;

[0062] (4) Screening, inoculate the separated enzymatic solution into the basal medium until cell clusters appear, observe the shape of the cells to determine the liver stem cells, and screen out the liver stem cells;

[0063] (5) Preservation. Store the liver stem cells in a low-temperature room at a temperature of 4°C...

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PUM

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Abstract

The invention discloses a preparation and extraction method of liver stem cells. The method comprises the following steps: cleaning and disinfecting, washing the surface layer of liver tissue by usingclean water, scrubbing by using an alcohol pad, and then dipping in normal saline for 3min; performing enzymolysis, taking out the liver tissue, then washing clean by using a PBS buffer solution, cutting the liver tissue into small pieces to put in a protease solution, dissolving for 6-24h, and filtering an enzymolysis solution by using a filter screen; separating; screening, inoculating a basalculture medium with the separated enzymolysis solution until a cell cluster appears, observing cell shapes to determine the liver stem cells, and screening the liver stem cells; preserving. The invention relates to the technical field of bioengineering. By the preparation and extraction method of the liver stem cells, the screening process of the stem cells is simpler, the screening cost of the stem cells is lowered, meanwhile, impurities which are possible to appear in stem cell tissue fluid are effectively removed, and the research and use value of the extracted stem cells is improved.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for preparing and extracting liver stem cells. Background technique [0002] Stem cells are a kind of pluripotent cells with self-replication ability. Under certain conditions, they can differentiate into a variety of functional cells. According to the developmental stage of stem cells, they can be divided into embryonic stem cells and adult stem cells. According to the developmental potential of stem cells, they can be divided into three types: Classes: totipotent stem cells, pluripotent stem cells and unipotent stem cells (multipotent stem cells), stem cells are not fully differentiated, immature cells with the potential to regenerate various tissues and organs and the human body. with cells". However, the screening process of stem cells in the existing stem cell extraction method is complicated, the screening cost is high, and impurities are easy to remain in ...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0672C12N2500/90C12N2509/00
Inventor 安徽
Owner 安徽
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