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Three-dimensional engineering breast cancer lung metastasis model, construction method and application

A technology of metastasis model and construction method, which is applied in the fields of biochemical equipment and methods, tumor/cancer cell, compound screening, etc., to achieve good application prospects and improve accuracy.

Active Publication Date: 2021-08-20
WEIFANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the 3D scaffolds based on natural and synthetic materials are not the same as the extracellular matrix of lung tissue in vivo, which leads to the difference between the 3D tissue engineering model of breast cancer lung metastasis and the real environment of lung metastasis in vivo, which makes the research and understanding of breast cancer difficult. After transfer to the lung, there are large discrepancies in the series of biological behaviors and mechanisms

Method used

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  • Three-dimensional engineering breast cancer lung metastasis model, construction method and application
  • Three-dimensional engineering breast cancer lung metastasis model, construction method and application
  • Three-dimensional engineering breast cancer lung metastasis model, construction method and application

Examples

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Embodiment 1

[0029] This embodiment provides a method for constructing a three-dimensional engineered breast cancer lung metastasis model, and the specific steps are as follows:

[0030] S1: Take pig-derived whole lung tissue, completely remove the bronchi, and grind the remaining part into 0.2cm 3 Wash the minced powder with PBS buffer solution with a pH of 7.4 for 5 times until the blood streaks are completely removed, and the washing time for each time is 0.5 h;

[0031] S2: Add SDS solution with a mass fraction of 0.5% to the minced powder treated in step S1 and stir for 10 hours to perform decellularization treatment;

[0032]S3: Add TritonX-100 solution with a volume fraction of 0.25% to the minced powder treated in step S2 and stir for 10 h to further remove cells, and then wash 5 times with PBS buffer solution with pH 7.4 to wash away residual For cell reagents, each washing time is 0.5h;

[0033] S4: Pre-freeze the minced powder obtained in step S3 at -20°C for 12 hours, then fr...

Embodiment 2

[0039] This embodiment provides a method for constructing a three-dimensional engineered breast cancer lung metastasis model, and the specific steps are as follows:

[0040] S1: Take pig-derived whole lung tissue, completely remove the bronchi, and grind the remaining part into 0.2cm 3 Wash the minced powder with PBS buffer solution with a pH of 7.4 for 5 times until the blood streaks are completely removed, and the washing time for each time is 0.5 h;

[0041] S2: Add SDS solution with a mass fraction of 0.5% to the minced powder treated in step S1 and stir for 10 hours to perform decellularization treatment;

[0042] S3: Add TritonX-100 solution with a volume fraction of 0.25% to the minced powder treated in step S2 and stir for 10 h to further remove cells, and then wash 5 times with PBS buffer solution with pH 7.4 to wash away residual For cell reagents, each washing time is 0.5h;

[0043] S4: Pre-freeze the minced powder obtained in step S3 at -20°C for 12 hours, then f...

Embodiment 3

[0050] In this example, the whole porcine lung tissue is excised from the bronchi, cut into small pieces, and crushed into pieces about 0.5 cm in size using a grinder. 3 Put the minced powder in a beaker, add 1000mL of ultrapure water, magnetically stir for 30min, replace with new ultrapure water, repeat three times, add 1000mL of PBS and wash for 3 times, magnetically stir for 30min each time, until there is no After obvious blood color, add 0.5% (wt%, the same below) SDS solution 1000mL, after magnetic stirring for 5h, replace the new 0.25% SDS solution and continue stirring for 5h to remove the cells; then, add 0.25% (v%, the same below ) Stir 1000 mL of TritonX-100 solution for 5 hours to further remove the remaining cells; after the cells are removed, add 1000 mL of PBS to wash 5 times, each time lasting 30 minutes, for washing the decellularization reagent. The decellularized powder was pre-frozen in a refrigerator at -20°C for 5 hours, and then dried in a freeze dryer a...

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Abstract

The invention relates to the field of tumor tissue engineering materials, and discloses a construction method of a three-dimensional engineering breast cancer lung metastasis model. The construction method includes construction of a primary diffuse leptomeningeal melanomatosis (PDLM)-collagen-chitosan scaffold, construction of a 3D breast cancer lung metastasis model of a single cell-composite scaffold, construction of a 3D breast cancer lung metastasis model of a multi-cell-composite scaffold and construction of a 3D breast cancer lung metastasis model of a multi-cell-composite scaffold-cytokine co-culture system. The invention further discloses the three-dimensional engineering breast cancer lung metastasis model and application. The model scaffold constructed by the invention is closer to components, mechanical strength and biochemical information of a lung extracellular matrix (ECM), the composite scaffold-based multi-cell co-culture system can better simulate a breast cancer lung metastasis microenvironment, constructed engineered tumor tissue is similar to biological characteristics of an in-vivo lung metastasis focus, and the model scaffold has important application value and prospect in research on the occurrence and development mechanism of breast cancer lung metastasis and an anti-tumor strategy.

Description

technical field [0001] The invention relates to the field of tumor tissue engineering materials, in particular to a three-dimensional engineered breast cancer lung metastasis model, a construction method and an application. Background technique [0002] Breast cancer is the malignant tumor with the highest incidence rate in women, and has become the number one killer of women's health. The distant metastasis of breast cancer is considered to be the direct cause of the high mortality rate and recurrence rate of breast cancer. It is the most likely target organ for metastasis. 60-70% of breast cancer patients have lung metastases in the end. As one of the main reasons affecting the survival of breast cancer patients, lung metastasis has become extremely important to study its occurrence, development and related molecular mechanisms. [0003] At present, it is extremely difficult to study lung metastasis of breast cancer in individual patients. In addition, there is a lack of ...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02
CPCC12N5/0693C12N5/0631G01N33/5011G01N33/5044C12N2513/00C12N2533/90C12N2533/54C12N2533/72C12N2501/15C12N2501/2306C12N2501/231C12N2503/02G01N2500/10
Inventor 李文芳孙同毅李磊王国娇王乐郑志高
Owner WEIFANG MEDICAL UNIV
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