Method for establishing colorectal cancer patient-derived xenograft model through three-dimensional culture system based on thermo-sensitive biogel

A xenotransplantation and colorectal cancer technology, applied in the field of biomedicine, can solve problems such as gene mutation, destruction of tumor tissue structure, and cell heterogeneity, and achieve the effects of simple preparation methods, reduced labor costs, and mature processes

Active Publication Date: 2018-06-12
ZHONGSHAN HOSPITAL FUDAN UNIV
View PDF5 Cites 12 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 Tiss

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for establishing colorectal cancer patient-derived xenograft model through three-dimensional culture system based on thermo-sensitive biogel
  • Method for establishing colorectal cancer patient-derived xenograft model through three-dimensional culture system based on thermo-sensitive biogel
  • Method for establishing colorectal cancer patient-derived xenograft model through three-dimensional culture system based on thermo-sensitive biogel

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] The results of a patient with colorectal cancer were cultured in a thermosensitive biogel three-dimensional culture system. The specific steps are as follows:

[0067] (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.

[0068] (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 to obtain the precipitated tissue fragments.

[0069] (3) Add 5ml of digestion solution, resuspend the pellet, shake vigorously, and incubate at 37°C, 5% CO2 incubator for 30min. After shaking vigorously again, add 5ml FCS (fe...

Embodiment 2

[0075]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:

[0076] (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.

[0077] (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 to obtain the precipitated tissue fragments.

[0078] (3) Add 5ml of digestion solution, resu...

Embodiment 3

[0085] 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:

[0086] (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.

[0087] (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 to obtain the precipitated tissue fragments.

[0088] (3) Add 5ml of digestion solution,...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to a three-dimensional culture system based on thermo-sensitive biogel. The three-dimensional culture system is prepared by combining the following reagents: thermo-sensitive biogel, digestive fluid required for separating and culturing tumor cells, a basal culture medium and a human intestinal stem cell culture medium. The invention also provides a method for establishing acolorectal cancer patient-derived xenograft model through the three-dimensional culture system based on thermo-sensitive biogel. The three-dimensional culture system has the advantages that colorectalcancer cells can be further amplified and grown in the three-dimensional culture system based on thermo-sensitive biogel, and the tumor formation capability of the colorectal cancer cells in the immune deficient mice can be enhanced. Polyclonal tumor cells cultured in the system have high subcutaneous tumor formation capability in nude mice. The three-dimensional culture system has the advantagesof simple preparation method and mature technology, and is capable of greatly reducing the time and the cost for establishing the human-derived colorectal cancer xenograft model.

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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
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
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap