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A method for establishing a xenograft tumor model derived from colorectal cancer patients based on a temperature-sensitive biogel three-dimensional culture system

A xenotransplantation and colorectal cancer technology, applied in the field of biomedicine, can solve the problems of destroying tumor tissue structure, cell heterogeneity, gene mutation, etc., achieve the effect of reducing labor cost, simple preparation method, and enhancing tumor formation ability

Active Publication Date: 2022-03-01
ZHONGSHAN HOSPITAL FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are many defects in the tumor model established by using cell lines: the process of establishing tumor cells in vitro is separated from the natural tumor microenvironment, resulting in mutations in their genes; selective pressure and digestive enzyme treatment during in vitro culture destroy the tumor Tissue structure, cell heterogeneity; animal model established by one cell line can only reflect one cancer patient
However, there is no report on a method of establishing a xenograft tumor model derived from colorectal cancer patients based on a thermosensitive biogel three-dimensional culture system of the present invention.

Method used

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  • A method for establishing a xenograft tumor model derived from colorectal cancer patients based on a temperature-sensitive biogel three-dimensional culture system
  • A method for establishing a xenograft tumor model derived from colorectal cancer patients based on a temperature-sensitive biogel three-dimensional culture system
  • A method for establishing a xenograft tumor model derived from colorectal cancer patients based on a temperature-sensitive biogel three-dimensional culture system

Examples

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Effect test

Embodiment 1

[0064] The results of a patient with colorectal cancer were cultured in a thermosensitive biogel three-dimensional culture system. The specific steps are as follows:

[0065] (1) Put the fresh colorectal cancer tissue (within 15 minutes from the body) surgically removed in a patient into a centrifuge tube containing 20ml DMEM (200U / ml penicillin / streptomycin), place it in an environment of 4°C, and transfer it rapidly Go to the ultra-clean table and set aside.

[0066] (2) Place the tumor tissue in a petri dish on ice, wash it with PBS until it is clean, and cut the tissue into 2-4mm 3 Fragments of different sizes were placed in a 15ml centrifuge tube (containing about 10ml of ice-cold PBS); centrifugation: 3min, 1500rpm; the supernatant was sucked off, and the tissue fragments were precipitated.

[0067] (3) Add 5ml of digestion solution, resuspend the pellet, shake vigorously, 37°C, 5% CO 2 Incubate in the incubator for 30min. After shaking vigorously again, add 5ml FCS (...

Embodiment 2

[0073] Based on the temperature-sensitive biogel three-dimensional culture system method compared with the subcutaneous embedding method of tissue pieces in 6 patients with colorectal cancer, a xenograft tumor mouse model derived from colorectal cancer patients was established. The establishment method is as follows:

[0074] (1) Put fresh colorectal cancer tissues (within 15 minutes from the body) of 6 patients into centrifuge tubes containing 20ml DMEM (200U / ml penicillin / streptomycin), place them in an environment of 4°C, and transfer them rapidly Go to the ultra-clean table and set aside.

[0075] (2) Place the tumor tissue in a petri dish on ice, wash it with PBS until it is clean, and cut the tissue into 2-4mm 3 Fragments of different sizes were placed in a 15ml centrifuge tube (containing about 10ml of ice-cold PBS); centrifugation: 3min, 1500rpm; the supernatant was sucked off, and the tissue fragments were precipitated.

[0076] (3) Add 5ml of digestion solution, resus...

Embodiment 3

[0083] Based on the three-dimensional culture system method of colorectal cancer cell biogel compared with the subcutaneous embedding method of tissue pieces in 6 patients with colorectal cancer, a xenograft tumor mouse model derived from colorectal cancer patients was established. The establishment method is as follows:

[0084] (1) Put fresh colorectal cancer tissues (within 15 minutes from the body) of 6 patients into centrifuge tubes containing 20ml DMEM (200U / ml penicillin / streptomycin), place them in an environment of 4°C, and transfer them rapidly Go to the ultra-clean table and set aside.

[0085] (2) Place the tumor tissue in a petri dish on ice, wash it with PBS until it is clean, and cut the tissue into 2-4mm 3 Fragments of different sizes were placed in a 15ml centrifuge tube (containing about 10ml of ice-cold PBS); centrifugation: 3min, 1500rpm; the supernatant was sucked off, and the tissue fragments were precipitated.

[0086] (3) Add 5ml of digestion solution,...

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Abstract

The invention relates to a three-dimensional culture system based on a temperature-sensitive biogel. The three-dimensional culture system is prepared by combining the following reagents: a temperature-sensitive biogel, a digestive juice required for separating and culturing tumor cells, a basal medium, Human intestinal stem cell culture medium. The invention also provides a method for establishing a xenograft tumor model derived from a colorectal cancer patient based on a temperature-sensitive biogel three-dimensional culture system. The advantages are shown in: colorectal cancer cells can be further expanded and grown in the temperature-sensitive biogel three-dimensional culture system of the present invention, and their tumorigenic ability in immunodeficient mice can be enhanced. The polyclonal tumor cells cultured in this system have good subcutaneous tumorigenic ability in nude mice. The preparation method of the invention is simple and the technology is mature, and the man-hour cost for the construction of the human-derived colorectal cancer xenograft tumor model can be greatly reduced.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a method for establishing a xenograft tumor model derived from colorectal cancer patients based on a temperature-sensitive biogel three-dimensional culture system. Background technique [0002] Colorectal cancer (colorecal cancer, CRC) is one of the most common malignant tumors worldwide, and its incidence ranks third in the incidence of malignant tumors, and the third in mortality. About 25% of patients with colorectal cancer are found to have synchronous liver metastases at the first diagnosis, and 25% to 50% of patients will develop metachronous liver metastases. The median survival time of patients with unresectable liver metastases is only 4.2 to 21.3 months. The 5-year survival rate after surgical resection of colon cancer liver metastases is nearly 40%. Combined with our previous clinical research findings, after chemotherapy and targeted therapy for patients with li...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09A01K67/027
CPCA01K67/0271C12N5/0693C12N2533/90C12N2513/00A01K2267/03A01K2227/105A01K2207/12
Inventor 许剑民何国栋鉴谧常文举任黎韦烨钟芸诗牛正川冯青阳朱德祥黄文柏刘天宇杨亮亮陈竟文易拓吉美玲彭上晋林奇
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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