Establishment and application of mmhrl3 transgenic mouse liver tumor cell line

A liver tumor and transgenic technology, applied in the fields of biology and oncology, can solve the problems of difficult establishment of mouse liver tumor cell lines, complicated influencing factors, time-consuming and labor-intensive, etc.

Inactive Publication Date: 2020-05-15
DALIAN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the high cost, time-consuming and labor-intensive experiments in mice, and the complex influencing factors, it is urgent to establish corresponding mouse liver tumor cell lines
However, in contrast, the establishment of mouse liver tumor cell lines is more difficult

Method used

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  • Establishment and application of mmhrl3 transgenic mouse liver tumor cell line
  • Establishment and application of mmhrl3 transgenic mouse liver tumor cell line
  • Establishment and application of mmhrl3 transgenic mouse liver tumor cell line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Establishment of Liver Tumor Cell MMHRL3 in H-ras12V Transgenic Male Mice

[0062] (a) source

[0063] H-ras12V transgenic male mouse liver tumor cell line comes from H-ras12V transgenic male mice, and its specific

[0064] The information is shown in the table below.

[0065] Table 1

[0066]

[0067] The primary liver tumor tissue of the H-ras12V transgenic male mouse was obtained, and the liver tumor tissue was directly cultured in vitro.

[0068] Cultured, repeatedly digested and passaged, and finally obtained a stable and passaged H-ras12V transgenic male mouse liver tumor cell line. Specifically include the following processes:

[0069] The above-mentioned H-ras12V transgenic male mice were sacrificed painlessly, and the liver tumors were excised by aseptic technique. Place the liver tumor in a 10cm culture dish filled with sterilized normal saline at normal temperature, fully clean the tumor capsule, and use a scalpel to cut the liver tumor tissue collect...

Embodiment 2

[0073] Biological characteristics of liver tumor cell MMHRL3 in H-ras12V transgenic male mice

[0074] In this example, conventional methods were used to detect the biological characteristics of the purebred H-ras12V transgenic male mouse liver tumor cell MMHRL3 with the preservation number CCTCC NO: C2016192, and the results are as follows:

[0075] 2.1 Cell morphology

[0076] Observing H-ras12V transgenic male mouse liver tumor cells MMHRL3 under an ordinary optical microscope, it can be seen that most of the liver tumor cells are round or polygonal, and the cell shape is uniform ( figure 2 A). Cell Counting Kit-8 (CCK8 kit) was used to detect the proliferation state of the cell line, and the operation was carried out in full accordance with the operation guide of the kit ( figure 2 B). The cells proliferated stably.

[0077] 2.2 Genotype identification of H-ras12V transgene

[0078] Take the H-ras12V transgenic male mouse liver tumor cell MMHR L3 (preservation numbe...

Embodiment 3

[0091] Establishment of liver tumor animal model using H-ras12V transgenic male mouse liver tumor cell MMHR1

[0092] The H-ra s1 2V transgenic male mouse liver tumor cell MMHR L3 (preservation number: C2016192) obtained in Example 1 was digested with digestive solution (0.25% trypsin, 0.02% EDTA, pH7.3), collected and count, according to 2 x 10 6 or 1×10 8 The number of cells / mouse was inoculated into the subcutaneous parts of the left and right axilla of the mice, and the inoculated mice were 5 4-week-old nude mice.

[0093] Between 15 and 40 days after inoculation, tumors occurred successively in 5 mice. The same method as in Example 2 was used for biological detection, and it was confirmed that hepatocellular liver tumors were induced in animal models.

[0094] Cell line deposit

[0095] The H-ras12V transgenic male mouse liver tumor cell MMHRL3 of the present invention was deposited in the China Center for Type Culture Collection (Wuhan, China) on November 08, 2016, a...

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Abstract

The invention provides establishment and application of an H-ras12V transgenic male mouse hepatoma cell line MMHRL3. Specifically, the character of the H-ras12V transgenic male mouse hepatoma cell line MMHRL3 is stable, passage can be achieved stably for multiple times, and the H-ras12V transgenic male mouse hepatoma cell line MMHRL3 is suitable for building a hepatoma animal model, and is an ideal cell line for basic and pre-clinical application research of liver cancer. Besides, the H-ras12V transgenic male mouse hepatoma cell line MMHRL3 expresses the H-ras mutation oncogene and has a highly activated Ras / MAPK and PI3K / AKT / mTOR signal channel. The invention further provides application of the cell line to animal model building and drug screening.

Description

technical field [0001] The present invention relates to the fields of biology and oncology, and more specifically relates to a novel H-ras12V transgenic male mouse liver tumor cell line MMHRL3, its establishment method and application. Background technique [0002] The occurrence and development of human diseases is a very complex process. In-depth study of the pathogenesis of various diseases and the effect of clinical drug treatment in order to achieve the purpose of preventing and curing various diseases is the unremitting goal of human beings. However, based on humanitarianism, direct experiments on patients cannot be used. In accordance with the Animal Protection Law, animal experiments can be used to conduct in-depth research on the occurrence and development of various diseases and life phenomena, so as to achieve the purpose of controlling the occurrence and development of human diseases, relieving patients' pain, and finally curing diseases. [0003] In the long-t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10A01K67/027A61K49/00
CPCA01K67/0271A01K2207/12A01K2227/105A01K2267/0331A61K49/0008C12N5/0693C12N2510/00
Inventor 王爱国樊婷婷刘要甫王靖宇王福金
Owner DALIAN MEDICAL UNIVERSITY
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