Three-dimensional drug resistance model of tumor cell chemotherapy drug and construction method thereof

A tumor cell, three-dimensional model technology, applied in the field of three-dimensional tumor cell culture, to achieve the effects of wide range of use, simple cell recovery, simple and mild preparation

Inactive Publication Date: 2019-05-10
SHENZHEN LUOHU PEOPLELS HOSPITAL
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Problems solved by technology

In addition, three-dimensional culture can simulate the microenvironment of tissue center necrosis and the interaction with extracellular matrix caused by hypoxia and lack of nutrients in tumor tissue in vivo,

Method used

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  • Three-dimensional drug resistance model of tumor cell chemotherapy drug and construction method thereof
  • Three-dimensional drug resistance model of tumor cell chemotherapy drug and construction method thereof

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Embodiment Construction

[0017] In order to facilitate the understanding of the technical solution of the present invention, the application of the three-dimensional drug resistance model of tumor cell chemotherapeutic drugs and its establishment method and the screening of anti-tumor drugs will be further described below in conjunction with specific examples.

[0018] 1. Construction of a three-dimensional drug resistance model of tumor cells

[0019] (1) Preparation: Bladder cancer cells were selected, and after the cells reached the logarithmic growth phase, they were digested with 0.25% trypsin-0.02% EDTA, collected by centrifugation, and counted. Centrifuge the cell suspension to remove the supernatant, add an appropriate amount of sterile sodium alginate solution (1.5%, w / v), and adjust the cell density of the solution to 1×10 6 cells / mL. The cell suspension was dropped into 10 times the volume of 100mM CaCl2 solution by hand-held injection, and calcified for 30 minutes to form calcium alginat...

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Abstract

The invention belongs to the field of three-dimensional tumor cell culture and particularly relates to a three-dimensional drug resistance model of a tumor cell chemotherapy drug and a construction method thereof. The three-dimensional drug resistance model is obtained by dropwise adding an alginate-bladder cancer cell suspension into a divalent/trivalent metal cation salt bath for chelation and preparing cell-loaded alginate gel microspheres for three-dimensional culture. The tumor cell mass obtained by three-dimensional culture has acquired chemotherapy drug resistance, has the performance that the activity of three-dimensional cells is remarkably higher than that of planar cells under the same drug concentration, meanwhile the expression level of an ABCG2 transporter is remarkably improved compared with the expression level of the planar cells, and the chemotherapy drug resistance cell model is successfully constructed. The anti-tumor drug high-throughput screening and evaluation model can be formed by adjusting three-dimensional culture conditions of the cells, meanwhile can also be used for researching the acquired drug resistance mechanism of clinical chemotherapy drugs of tumor cells and find the target of reversal of the clinical chemotherapy drug resistance of bladder cancer cells and has a good application value.

Description

technical field [0001] The invention belongs to the field of three-dimensional culture of tumor cells, and specifically relates to a three-dimensional drug resistance model of tumor cells for chemotherapeutic drugs, its establishment method and the application of anti-tumor drug screening. Background technique [0002] To study the regulatory effect of other cells in the tumor microenvironment on the invasion and metastasis of tumor cells in vitro, it is necessary to construct a culture model that can simulate the tumor microenvironment in vivo. Although the traditional planar cell culture method is a common technical means for tumor research in vitro, it cannot simulate the complexity of the in vivo microenvironment, resulting in the loss of tissue-specific structure of cells, affecting the expression and conduction of biomechanical and biochemical signals, Affect intercellular connections, etc., so that the results obtained in planar culture are different from those in viv...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02
Inventor 吴松刘晓岑
Owner SHENZHEN LUOHU PEOPLELS HOSPITAL
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