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Anti-tumor drug screening kit based on three-dimensional micro-tissue level and application method thereof

An anti-tumor drug, micro-tissue technology, applied in biochemical equipment and methods, tumor/cancer cell, microorganism determination/inspection, etc., can solve cell signal transduction, abnormal changes in protein expression, loss of biological characteristics of cells, Inconvenient clinical application and other problems, to achieve reliable drug susceptibility testing data, achieve drug susceptibility assessment, and short time-consuming effects

Inactive Publication Date: 2018-01-09
宋焱艳
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Second, patients have individual differences, and different individuals will show different sensitivities to different drugs
2D cultured cells grow in a single layer of adherent cells, and their morphology and the way cells interact with the environment are different from those in vivo, which leads to abnormal changes in cell signal transduction and protein expression, and eventually causes cells cultured in vitro to gradually lose their original The biological characteristics of the experimental results obtained on this basis are often difficult to be verified in vivo experiments
Chinese patent CN104111254A provides a method for culturing cells in collagen for drug susceptibility testing of tumors, but it is inconvenient for clinical application because of cumbersome operation steps

Method used

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  • Anti-tumor drug screening kit based on three-dimensional micro-tissue level and application method thereof
  • Anti-tumor drug screening kit based on three-dimensional micro-tissue level and application method thereof
  • Anti-tumor drug screening kit based on three-dimensional micro-tissue level and application method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] A three-dimensional micro-tissue level personalized drug screening kit, including the following components:

[0051] (1) Three-dimensional cell culture material: microporous 3D suspension culture plate.

[0052] (2) Tissue cell dispersing reagent: 0.05% trypsin-EDTA, refrigerated at -20°C.

[0053] (3) Cell culture medium: RMPI 1640 cell culture medium.

[0054] (4) Fetal bovine serum. Serum was added to basal cell culture medium at 10% by volume prior to use. Serum needs to be refrigerated at -20°C.

[0055] (5) Balanced salt solution: PBS.

[0056] (6) 100╳ antibiotics: 10000U / ml penicillin chain and 10mg / ml streptomycin mixture. Refrigerate at -20°C. Add to medium before use and dilute to 1╳ working concentration.

[0057] (7) Cell viability detection reagent: alamarBlue.

[0058] (8) Cell strainer: sterile plastic cell strainer with a pore size of 100 μm.

[0059] A method for using a three-dimensional microtissue-level personalized drug screening kit in br...

Embodiment 2

[0088] A three-dimensional micro-tissue level personalized drug screening kit, including the following components:

[0089] (1) Three-dimensional cell culture material: 96-well cell culture plate and 3D cell culture hydrogel.

[0090] (2) Tissue cell dispersing reagent: 0.05% trypsin-EDTA, 1000 U / mL collagenase. Refrigerate at -20°C.

[0091] (3) Cell culture medium: RMPI 1640 cell culture medium.

[0092] (4) Fetal bovine serum. Serum was added to basal cell culture medium at 10% by volume prior to use. Serum needs to be refrigerated at -20°C.

[0093] (5) Balanced salt solution: PBS.

[0094] (6) 100╳ antibiotics: 10000U / ml penicillin chain and 10mg / ml streptomycin mixture. Refrigerate at -20°C. Add to medium before use and dilute to 1╳ working concentration.

[0095] (7) Cell viability detection reagent: alamarBlue.

[0096] (8) Cell strainer: sterile plastic cell strainer with a pore size of 100 μm.

[0097] A method for using a three-dimensional microtissue-level ...

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Abstract

The invention relates to an anti-tumor drug screening kit based on a three-dimensional micro-tissue level and an application method thereof. The anti-tumor drug screening kit comprises a three-dimensional cell culture material, a tissue cell dispersion reagent, a proteinase inhibitor, a basic cell culture medium, blood serum, antibiotics, a balance salt solution, a cell activity detection reagentand a cell filter screen. The application method comprises the following steps: dispersing tumor tissues of a patient to obtain tissue blocks, tissue fragments, cell aggregates and / or dispersed singlecells; then culturing the dispersed tumor tissues of the patient and a cell sample through a three-dimensional culture method and enabling the cells to grow in a three-dimensional space to form a spherical structure or an irregular three-dimensional structure with a stereoscopic shape; finally, carrying out a drug sensitivity test on tumor micro-tissues obtained by three-dimensional culture. Theanti-tumor drug screening kit based on the three-dimensional micro-tissue level is simple to operate and is suitable for the drug sensitivity tests of various solid tumors.

Description

technical field [0001] The invention relates to a tumor individualized drug screening kit based on three-dimensional micro-tissue level and a method for using the same. Specifically, it is composed of a three-dimensional micro-tissue-level tumor personalized drug screening kit. The kit is used to collect and three-dimensionally culture tumor cells of a patient to perform drug sensitivity testing. The kit obtains tumor micro-tissues by three-dimensional culture method, which can maintain the biological characteristics of tumor cells to the greatest extent in in vitro experiments, and can obtain reliable drug susceptibility detection data, which can provide reference for the formulation of personalized treatment plans for clinical tumors. Improve the accuracy of medication. Background technique [0002] At present, there are many kinds of anti-tumor drugs commonly used in clinic, but the efficacy is still not ideal. Because, first, there is heterogeneity in the tumor itself,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02C12N5/09
Inventor 宋焱艳
Owner 宋焱艳
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