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Human well-differentiated liver cancer cell line HL1017 and construction method thereof

A technology of liver cancer cells and a construction method, which is applied to the construction of the cell line, the field of human-derived well-differentiated liver cancer cell line HL1017, can solve the problems of poor conventional medical treatment, insufficient patient ratio, and insufficient liver source, etc. Conducive to the maintenance of cell growth and function, reducing safety concerns, and the effect of good cell growth

Inactive Publication Date: 2013-09-25
NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002]Hepatic failure (hepatic failure) is a serious clinical syndrome, the effect of conventional medical treatment is not good, and the mortality rate is as high as 70%
Liver transplantation is the only effective treatment for advanced liver failure, but there is a serious shortage of liver sources at present, and the proportion of patients who can receive liver transplantation is less than 1%
However, there are some deficiencies and defects in these technologies and products, and foreign companies enjoy patent protection for some products

Method used

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  • Human well-differentiated liver cancer cell line HL1017 and construction method thereof
  • Human well-differentiated liver cancer cell line HL1017 and construction method thereof
  • Human well-differentiated liver cancer cell line HL1017 and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] 1. Isolation and culture of primary human liver cells

[0058] For the surgically resected liver cancer marginal tissue, the primary liver cells were separated by the collagenase two-step perfusion method of small pieces of liver tissue established in our laboratory. The specific steps are as follows:

[0059] (1) The specimens were obtained from fresh isolated adult liver cancer tissue resected by hepatobiliary surgery in Nanfang Hospital. The liver cancer tissue was pathologically diagnosed as well-differentiated liver cancer tissue. Remove the connective tissue of the liver cancer tissue, use a 5ml sterile syringe to draw HBSS balanced salt solution, and perform multi-point puncture and perfusion on the liver cancer tissue until the liver cancer tissue turns pale.

[0060] (2) Use a 5ml sterile syringe to draw 0.05% type IV collagenase solution preheated at 37°C for the second multi-point puncture and perfusion until the liver cancer tissue becomes soft and inelast...

Embodiment 2

[0162] Three-dimensional culture of HL1017 in hydrogel

[0163] 1) Take 20μL Matrigel and evenly pre-embed the bottom of the 24-well plate, place at 37°C for about half an hour until it solidifies;

[0164] 2) Digestion of HL1017 cells

[0165] ① Remove the old culture medium in the culture bottle;

[0166] ② Add 2ml of PBS, shake the culture flask gently, so that the PBS flows over all the cell surfaces, then remove the PBS, and repeat the operation 1 or 2 times;

[0167] ③ Add 1ml of 0.25% trypsin digestion solution (0.25% trypsin and 0.02% EDTA), shake the culture bottle gently, so that the trypsin digestion solution flows over all the cell surfaces, let it stand for 2~3min, and observe the cell changes under an inverted microscope. After the circle floats up and the intercellular space increases, remove the digestion solution and upright the culture bottle, add a culture solution equal to the volume of trypsin to stop the digestion;

[0168] ④ Use a pipette to gently bl...

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Abstract

The present invention discloses a human well-differentiated liver cancer cell line HL1017, wherein a preservation number is CCTCC NO:C201357, and the cell line has the most important functions of synthesis, metabolism and detoxification of liver. In addition, the present invention further discloses a construction method of the human well-differentiated liver cancer cell line HL1017.

Description

technical field [0001] The invention belongs to the technical field of biological materials, and in particular relates to a human-derived well-differentiated liver cancer cell line HL1017, and also relates to a construction method of the cell line. Background technique [0002] Hepatic failure is a serious clinical syndrome, the effect of conventional medical treatment is not good, and the mortality rate is as high as 70%. Liver transplantation is the only effective treatment for advanced liver failure, but at present there is a serious shortage of liver sources, and the proportion of patients who can receive liver transplantation is less than 1%. The emergence of bioartificial liver (BAL) provides a glimmer of hope for the treatment of liver failure. It can temporarily assist and partially replace the function of the diseased liver, and help patients with liver failure recover their own liver function or make them wait until liver transplantation. [0003] However, the bio...

Claims

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Application Information

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IPC IPC(8): C12N5/09
Inventor 刘思德燕群李爱民邓丽娟王亚东
Owner NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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