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Inoculation liquid composition and method for building malignant pleural effusion sourced xenotransplantation tumor animal model by using inoculation liquid composition

The technology of a composition and inoculation solution is applied in the field of construction of xenograft tumor animal models, which can solve the problems of inability to construct xenograft tumor animal models of malignant pleural effusion, kill tumor cells, etc., achieve simple establishment process, increase success rate, The effect of retaining the biological activity of cells

Active Publication Date: 2017-05-31
SHANGHAI CHEST HOSPITAL +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the number of tumor cells in malignant pleural effusion is not only small, but also contains dead or near-dead tumor cells, pleural effusion also contains immune cells and other cells, inflammatory factors produced by immune cells and their itself may kill tumor cells
Due to the above limitations, it is generally believed that malignant pleural effusion cannot be used to establish xenograft tumor animal models

Method used

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  • Inoculation liquid composition and method for building malignant pleural effusion sourced xenotransplantation tumor animal model by using inoculation liquid composition
  • Inoculation liquid composition and method for building malignant pleural effusion sourced xenotransplantation tumor animal model by using inoculation liquid composition
  • Inoculation liquid composition and method for building malignant pleural effusion sourced xenotransplantation tumor animal model by using inoculation liquid composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] Example 1 Construction of xenograft tumor mouse model derived from malignant pleural effusion of lung cancer

[0080] 1. experimental animals

[0081] Order 4-5-week-old female severe combined immunodeficiency mice from the supplier and keep them in an SPF-level animal room. The animals are adaptively fed for at least three days before the start of the experiment.

[0082] 2. experiment procedure

[0083] 2.1 Collection of clinical malignant pleural effusion samples for lung cancer

[0084] Collect 200ml freshly collected malignant pleural effusion samples from non-small cell lung cancer in test tubes containing anticoagulants, transport them on ice, and transport them to the laboratory within 24 hours. A total of 15 cases of malignant pleural effusion were collected.

[0085] 2.2 Enrichment of tumor cells in malignant pleural effusion

[0086] Dilute the malignant pleural effusion sample with one volume of PBS buffer (HyClone, SH30256.01) in a biological safety...

Embodiment 2

[0097] Example 2 Drug efficacy experiment carried out on non-small cell lung cancer model CTC15012

[0098] In order to verify that the lung cancer mutation model established by the present invention is consistent with the clinical response, corresponding drug efficacy experiments consistent with the drugs taken by clinical patients are carried out on the established animal model.

[0099] Female NU / NU nude mice were subcutaneously inoculated with human ALK fusion gene model CTC15012 tumor mass. The non-small cell lung cancer model CTC15012 was derived from a 55-year-old female patient whose pathological diagnosis was bronchopulmonary adenocarcinoma. Cut the tumor into small pieces in HBSS and inoculate it under the skin of nude mice until the tumor grows to 500-700mm 3 When , the tumor was cut into small pieces of 10-15 mg in HBSS culture medium and used for subcutaneous inoculation in 20 experimental nude mice. The specific inoculation method is: disinfect the skin on both...

Embodiment 3

[0104] Example 3 Drug efficacy experiment carried out on non-small cell lung cancer model CTC15008

[0105] Female NU / NU nude mice were subcutaneously inoculated with human EGFR exon 21 L858R mutation model CTC15008 tumor mass. The non-small cell lung cancer model CTC15008 was derived from a 71-year-old female patient whose pathological diagnosis was bronchopulmonary adenocarcinoma. Cut the tumor into small pieces in HBSS and inoculate it under the skin of nude mice until the tumor grows to 500-700mm 3 When , the tumor was cut into small pieces of 10-15 mg in HBSS culture medium and used for subcutaneous inoculation in 20 experimental nude mice. The specific inoculation method is: disinfect the skin on both sides of the back of the mouse with povidone iodine, and use a No. 20 trocar to inject a total amount of 2 × 2 × 2mm 3 The tumor tissue is pierced on both sides of the back, the needle travels between the skin and the muscle, and finally the tissue is sent under the skin ...

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Abstract

The invention relates to an inoculation liquid composition and a method for building a malignant pleural effusion sourced xenotransplantation tumor animal model by using the inoculation liquid composition. The inoculation liquid provided by the invention contains the following ingredients including Hank' blanace salt buffer solution, fetal calf serum, basement membrane substrate glue and ROCK protein inhibitor. The method for building the malignant pleural effusion sourced xenotransplantation tumor animal model provided by the invention comprises the steps of collecting a clinic malignant pleural effusion sample, enriching tumor cells, using inoculation liquid suspension cells provided by the invention, and inoculating the cells into an animal so as to form xenotransplantation tumor. The model building method provided by the invention has the advantages that simplicity is easy; the implementation is easy; the tumor forming rate is high, so that the method can be used for medicine screening and mechanism study of various kinds of cancer with malignant pleural effusion.

Description

technical field [0001] The invention belongs to the field of medical models. More specifically, the present invention relates to the establishment of xenograft tumor animal models derived from malignant pleural effusion. Background technique [0002] In my country, lung cancer has replaced liver cancer as the first cause of death from malignant tumors, and the morbidity and mortality continue to rise rapidly. The high mortality rate of lung cancer is mainly due to the fact that 60-70% of lung cancer patients are found to be in the advanced stage of lung cancer, and they have lost the chance of operation and cannot be cured. [0003] Tumor animal models are an important platform for research on tumor-related mechanisms and drug screening. At present, preclinical tumor animal models mostly adopt the method of subcutaneous inoculation of tumor cell lines, which has the advantages of easy establishment and repeatability. However, due to the establishment of tumor cell lines i...

Claims

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Application Information

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IPC IPC(8): C12N5/09A01K67/027A61K49/00
CPCA01K67/0271A01K2207/12A01K2227/105A01K2267/0393A61K49/0008C12N5/0693C12N2500/60C12N2500/84C12N2501/727
Inventor 陆舜闻丹忆徐云华张菲菲徐文华杨阳陈智伟
Owner SHANGHAI CHEST HOSPITAL
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