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Construction method of magnetic control in-vitro tumor array

A construction method and magnetron technology, applied in the field of construction of tumor arrays in vitro, can solve the problems of high cost, complicated operation, difficult high throughput, etc., and achieve the effect of low production cost and simple operation

Active Publication Date: 2021-03-02
HARBIN INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to provide a method for constructing a magnetically controlled in vitro tumor array in order to solve the problems of difficulty in high-throughput production of tumor spheres with uniform size, complex operation and high cost in the prior art.

Method used

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  • Construction method of magnetic control in-vitro tumor array
  • Construction method of magnetic control in-vitro tumor array
  • Construction method of magnetic control in-vitro tumor array

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Experimental program
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specific Embodiment approach 1

[0019] Specific implementation mode 1: This implementation mode records a method for constructing a magnetically controlled tumor array in vitro, and the specific steps of the method are:

[0020] Step 1: Simple device preparation and cleaning: ultrasonically clean the braided nickel mesh, the container with the upper opening, and the NdFeB magnet in detergent, distilled water and absolute ethanol for 10-30 minutes, blow dry with nitrogen, and then fix the nickel mesh Put it on the bottom of the container and fix it in the center of the magnet, and finally put it under the ultra-clean table UV lamp for 2-4 hours; the nickel mesh is magnetic, and after placing a round magnet at the bottom, the nickel mesh and the magnet will attract each other, so the container can be fixed.

[0021] Step 2: Preparation of cell solution: collect the pre-cultured tumor cells in a centrifuge tube, concentrate to 1×10 8 / mL, take 3mL of the above cell suspension and mix evenly with 50-100μL gadobu...

specific Embodiment approach 2

[0024] Embodiment 2: The method for constructing a magnetron tumor array in vitro described in Embodiment 1, in step 1, the radiation intensity of the ultraviolet lamp is >100 μW / cm 2 . Ultraviolet lamp irradiation plays a role in sterilization and prevents the subsequent cultured tumor spheres from being infected with bacteria.

specific Embodiment approach 3

[0025] Embodiment 3: In the method for constructing a magnetically controlled in vitro tumor array described in Embodiment 1, in step 2, the tumor cells are brain glioma cells C6 or liver cancer cells Hepg2.

[0026] Embodiment 4: The method for constructing a magnetically controlled in vitro tumor array described in Embodiment 1, in step 3, the cell culture medium contains 50 μg / mL penicillin, 50 μg / mL streptomycin and 10% fetal bovine serum DMEM, the culture fluid provides energy to the cells.

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Abstract

The invention discloses a construction method of a magnetic control in-vitro tumor array, belongs to the field of biological medicines. and aims to solve the problems that a relatively uniform tumor ball is difficult to produce from a three-dimensional tumor ball and the operation is complex. The method comprises the following steps: ultrasonically cleaning and blow-drying a woven nickel net, a container and a neodymium iron boron magnet in a detergent, distilled water and absolute ethyl alcohol in sequence, fixing the nickel net at the bottom of the container, then fixing the nickel net at the center of the magnet, and irradiating under an ultraviolet lamp for 2-4 hours; tcollecting tumor cells in a centrifuge tube, carrying out centrifugal concentration, and mixing 3mL of cell suspensionwith gadobutrol; adding 100-200 [mu] L of the cell culture solution into a container, wetting a nickel net, adding 1-2 mL of the mixed solution into the container, and allowing cells to form a cell aggregate array on the nickel net; transferring the device into a cell culture box, replacing the cell culture solution every 1-2 hours, and forming a tumor array after 24-48 hours. The method is easyto operate and low in manufacturing cost, and can be used for producing tumor balls with uniform sizes in a high-throughput manner. The prepared tumor ball can be used for medical experiments.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a method for constructing a magnetically controlled tumor array in vitro. Background technique [0002] Scientists have found that there is still a big gap in the use of two-dimensional tumor cells to mimic tumor tissue and tumor microenvironment in vivo. Evidence shows that the gene expression levels in three-dimensional cultures in vitro can more accurately reflect the real tumor than those observed in two-dimensional cultures, and the establishment of three-dimensional tumors in vitro overcomes the shortcomings of animal models that cannot control a single variable , so tumorspheres are recognized as one of the best cell models for tumor research and have great application potential in the development of new anticancer therapeutic strategies. Regarding the establishment of three-dimensional tumor spheres, there are several relatively mature methods, such as suspension c...

Claims

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Application Information

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IPC IPC(8): C12N5/09C40B40/02C40B50/06
CPCC12N5/0693C40B40/02C40B50/06C12N2511/00
Inventor 韩晓军张祥祥
Owner HARBIN INST OF TECH