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A mouse model of circulating tumor cells, its construction method and application

A technology of tumor cells and mouse models, applied in the field of animal genetic engineering

Active Publication Date: 2021-04-13
湖南昭泰生物医药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, the methods of labeling, identifying and sorting human tumor CTC cells from human tumor mouse models also have the above problems

Method used

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  • A mouse model of circulating tumor cells, its construction method and application
  • A mouse model of circulating tumor cells, its construction method and application
  • A mouse model of circulating tumor cells, its construction method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1: Construction of lung cancer PDX model

[0064] 1) Treatment of primary lung cancer samples: Primary lung cancer tumor tissue samples were obtained from the tumor tissue sample bank of a tertiary hospital. Lung cancer samples should be soaked in RMPI-1640 medium and stored on ice for transportation. After obtaining the tumor sample, wash it 2-3 times with PBS (1% P / S) and divide the sample into three parts, one for cryopreservation; one for tumor tissue section and immunohistochemical analysis; one for tumor transplantation spare.

[0065] 2) Lung cancer tissue transplantation and detection: The appropriate tumor tissue transplantation method can be selected according to the size of the liver cancer sample. The transplant method is as follows:

[0066] (1) Subcutaneous transplantation: cut the primary tumor tissue sample into 15mm with scissors 3 Small pieces of tumor tissue, the small pieces of lung cancer tumor tissue were wrapped with Matrix Gel for l...

Embodiment 2

[0068] Example 2: Multiple passages of lung cancer PDX model

[0069] Lung cancer PDX mouse model passage: the lung cancer PDX mice of Example 1 were passed CO 2 After sacrifice, the mouse tumor tissue was taken, and the flesh-colored part of the outer layer of the tumor tissue was cut off, washed 2-3 times with PBS (1% P / S), and the sample was divided into three parts, one part was frozen for preservation; one part was used for tumor Tissue section and immunohistochemical analysis; one copy for tumor transplantation;

[0070] (1) Subcutaneous transplantation: cut the primary tumor tissue sample into 15mm with scissors 3 Small pieces of tumor tissue, the small pieces of lung cancer tumor tissue were wrapped with Matrix Gel for later use, a small opening was cut on the side of the lower abdomen of the mouse with scissors, the tumor tissue piece wrapped in MatrixGel was clamped into the small opening with tweezers, and the small opening was closed with wound clips. suture;

...

Embodiment 3

[0073] Embodiment 3: Detection of lung cancer PDX model

[0074] Within 30-60 days of tumor transplantation, CO 2 The PDX model of embodiment 1 and embodiment 2 was put to death, and the mouse peripheral blood, lung, liver, spleen and tumor tissue were taken, and the proportion (hHLA+) and migration (except tumor tissue) of tumor cells therein were detected by flow cytometry. organ hHLA+ cell ratio), the results are shown in Figure 1-2.

[0075] Result analysis: From the results of immunohistochemistry in Figure 1 (A), it was shown that tumors in PDX mice and patient tumors expressed consistent tumor markers (thyroid transcription factor 1 (TTF1), cytokeratin 7 (CK7), aspartate Acid protease A (NapsinA), indicating that after passage in immunodeficient mice, the tumor retains the original tissue structure characteristics. Figure 1 (B) immunohistochemical results show that lung cancer can metastasize to the lung, spleen and For major organs such as the liver, it can be seen t...

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Abstract

The present invention relates to a mouse model of circulating tumor cells, its construction method and application. The method comprises the following steps: (1) subcutaneous transplantation: transplanting primary tumor tissue samples into the body of immunodeficiency mice to construct primary cancer cells Xenograft model PDX; (2) collection of circulating tumor cells: collection of circulating tumor cells in the peripheral blood of the primary cancer xenograft model described in step (1); (3) kidney capsule transplantation: collection of step (2) The circulating tumor cells were transplanted into the kidney capsule of immunodeficient mice, and the mouse model of circulating tumor cells was successfully constructed. The method of the present invention adopts the method of more than two times of multi-passage transplantation, and transplants the CTCs in the primary tumor xenograft model into the body of an immunodeficiency mouse through the kidney capsule to obtain a mouse model of circulating tumor cells, which can be used to study CTCs in vivo Mechanism of transfer and spread.

Description

technical field [0001] The invention belongs to the technical field of animal genetic engineering, and in particular relates to a circulating tumor cell mouse model, its construction method and application. Background technique [0002] Lung cancer is the cancer with the highest death rate in China, and the 5-year survival rate of lung cancer patients is only 8%-10%. In recent years, many new discoveries have been made in the diagnosis and treatment of lung cancer. At present, surgical treatment, chemotherapy and radiotherapy can get good results for early lung cancer. However, these methods are not very suitable for patients with stage IV lung cancer that has been widely metastasized. The dissemination and metastasis of lung cancer is an important factor leading to the high mortality rate of lung cancer. Therefore, research on the mechanism and process of lung cancer metastasis is an urgent need for the current lung cancer treatment research and development platform. [0...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09
CPCA01K67/0271A01K2207/12A61D7/00A61K35/13C12N5/0693C12N2509/00G01N33/57484
Inventor 不公告发明人
Owner 湖南昭泰生物医药有限公司
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