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Circulating tumor cell mouse model and construction method and application thereof

A tumor cell, mouse model technology, applied in the field of animal genetic engineering

Active Publication Date: 2017-09-08
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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  • Circulating tumor cell mouse model and construction method and application thereof
  • Circulating tumor cell mouse model and construction method and application thereof
  • Circulating tumor cell mouse model and construction method and application thereof

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 invention relates to a circulating tumor cell mouse model and a construction method and application thereof. The method includes the following steps of 1, subcutaneous transplantation, wherein a primary tumor tissue sample is transplanted into the body of an immunodeficient mouse to construct a primary cancer heterotransplantation model PDX; 2, collection of circulating tumor cells, wherein the circulating tumor cells in peripheral blood in the primary cancer heterotransplantation model obtained in the step 1 are collected; 3, renal sac membrane transplantation, wherein the circulating tumor cells collected in the step 2 are transplanted into the renal sac membrane of the immunodeficient mouse, and then the circulating tumor cell mouse model is successfully constructed. According to the method, the mode of transplantation with two or more times of passages is adopted, the CTCs in the primary cancer heterotransplantation model are transplanted into the body of the immunodeficient mouse through renal sac membrane, and the circulating tumor cell mouse model is obtained; the circulating tumor cell mouse model can be applied to research on the in-vivo metastasis mechanism and proliferation condition of CTCs.

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 Applications(China)
IPC IPC(8): A61K35/13A61D7/00C12N5/09G01N33/574A01K67/027
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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