Three-dimensional lung cancer model support, preparation method and application

A lung cancer and model technology, applied in biochemical equipment and methods, respiratory/lung cells, tumor/cancer cells, etc., can solve the problems of large demand for decellularization reagents, poor mechanical properties of natural materials, and lack of flexibility in operation. Achieve the effects of accurately predicting the curative effect in vivo, good proliferation space, and good permeability of the stent

Inactive Publication Date: 2021-08-24
WEIFANG MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The scaffold materials required for the construction of 3D tissue engineering tumor models mainly include naturally derived biomaterials and artificially synthesized polymer materials. Naturally derived materials, especially substances such as polysaccharides or proteins, have good biocompatibility and are conducive to the growth of cells. Adhesion and proliferation, but most natural materials have poor mechanical properties, and often need to be cross-linked or compounded with other materials for the construction of tissue engineering tumors
This method often requires a pe

Method used

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  • Three-dimensional lung cancer model support, preparation method and application
  • Three-dimensional lung cancer model support, preparation method and application
  • Three-dimensional lung cancer model support, preparation method and application

Examples

Experimental program
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Embodiment 1

[0037] This embodiment provides a method for preparing a three-dimensional lung cancer model bracket, which specifically includes the following steps:

[0038] S1: Take pig-derived whole lung tissue, wash it with pure water for 3-5 times, completely remove the bronchial part, and grind the remaining part into 0.2cm 3 ~0.5cm 3 the crumbs;

[0039] S2: Wash the mince in step S1 with phosphate (PBS) buffer solution with a pH of 7.4 at room temperature for 3 to 5 times to remove the blood streaks. Stir with a magnetic stirrer for each wash. 0.5~1h;

[0040] S3: Add an aqueous solution of sodium dodecylsulfonate (SDS) with a mass fraction of 0.1-0.5% to the minced powder treated in step S2 at room temperature, stir with a magnetic stirrer for 5-10 hours, and initially remove cells, Partially decellularized lung fragments were obtained;

[0041] S4: Add a TritonX-100 solution with a volume fraction of 0.1-0.25% to the minced powder treated in step S3 at room temperature, stir wi...

Embodiment 2

[0053] In this example, the whole porcine lung tissue is excised from the bronchi, cut into small pieces, and crushed with a grinder, with a size of about 0.5cm 3 , put the minced pig lungs in a beaker, add 1000mL of ultrapure water, stir magnetically for 30min, then replace with new ultrapure water, repeat three times, add 1000mL of PBS to wash 3 times, magnetically stir for 30min each time, until there is no obvious After the blood color, add 1000mL of 0.5% (wt%, the same below) SDS solution, and after magnetic stirring for 5h, replace with a new 0.25% SDS solution and continue stirring for 5h to remove cells; then, add 0.25% (v%, the same below) Stir 1000mL of TritonX-100 solution for 5h to further remove remaining cells; after the cells are removed, add 1000mL of PBS to wash 5 times, each time lasting 30min, 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 at -80°C f...

Embodiment 3

[0062] In this example, the whole porcine lung tissue is excised from the bronchi, cut into small pieces, and crushed to a size of about 0.2 cm using a grinder. 3 ; After that, put the minced pig lungs in a beaker, then add 1000mL ultrapure water to the beaker, and replace with new ultrapure water after magnetic stirring for 30 minutes. , after no obvious blood color in the sample, add 0.25% (wt%, the same below) SDS solution 1000mL, after magnetic stirring for 3 hours, replace with new 0.1% SDS solution and continue stirring for 3 hours to remove cells. Subsequently, add 1000 mL of 0.1% (v%, the same below) TritonX-100 solution and stir for 8 hours to further remove the remaining cells. After the cells are removed, add 1000 mL of PBS to wash 3 times, each time lasting 1 hour, for washing the decellularization reagent . Then, the decellularized powder was pre-frozen in a refrigerator at -30°C for 10 hours, and then dried in a freeze dryer at -75°C for 15 hours to obtain a dri...

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Abstract

The invention relates to the field of tumor tissue engineering materials, and discloses a preparation method of a three-dimensional lung cancer model scaffold, which comprises pig-derived whole lung tissue pretreatment, decellularized powder freeze-drying, nano-particle ball milling and secondary freeze-drying. The invention further discloses a three-dimensional lung cancer model support. Applications are also disclosed. The operation process is simple and flexible, the decellularization effect is good, the prepared composite scaffold simulates lung extracellular matrix parts as much as possible, influences of matrixes on tumor cell biological behaviors are better reflected, the scaffold is low in immunogenicity and good in biocompatibility, growth and proliferation of tumor cells are facilitated, transmission of nutrient substances and metabolic waste is facilitated, and the scaffold has good application prospects. A good proliferation space is provided for cell growth; the biological behaviors of cancer cells in the composite scaffold can be conveniently and intuitively researched, and a three-dimensional lung cancer model constructed by utilizing the composite scaffold can be used as an important research platform for screening anti-tumor drugs.

Description

technical field [0001] The invention relates to the field of tumor tissue engineering materials, in particular to a three-dimensional lung cancer model bracket, a preparation method and an application. Background technique [0002] At present, research models of lung cancer biology mainly include in vitro 2D cell culture models and in vivo animal models. In vitro 2D cell culture models lack the internal microenvironment and physiological correlation of tumors in vivo, and cannot reflect the high complexity, heterogeneity and biological relevance of tumor tissues. In vivo animal models have problems such as high cost of animal experiments, long cycle, complicated operation, ethical issues, and individual differences. Therefore, both in vitro 2D cell culture models and in vivo animal models have certain limitations for existing research and cannot meet requirements of modern medicine. The 3D culture model of tumor cells can simulate the growth microenvironment of tumor cells ...

Claims

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Application Information

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IPC IPC(8): C12N5/09
CPCC12N5/0693C12N5/0688C12N2513/00C12N2533/90C12N2533/54C12N2533/72
Inventor 李文芳孙同毅李磊王丹王乐窦泽民
Owner WEIFANG MEDICAL UNIV
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