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Tumor organoid preparation method and device and application thereof

An organoid and tumor technology, applied in the field of tumor organoid preparation, can solve problems such as difficulty in meeting clinical applications and scientific research, destroying the connection between cells, and a small number of organoids, achieving high value-added ability, high organoid proliferation ability, and improved Effects of Survival and Quantity

Active Publication Date: 2020-09-01
奥格诺德生物科技(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the enzymatic hydrolysis method to digest tissues and destroy intercellular connections, and the digestion time is longer, which reduces cell activity, the success rate of sample processing is low, and the number of organoids obtained is small, which is difficult to meet the needs of clinical applications and scientific research.

Method used

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  • Tumor organoid preparation method and device and application thereof
  • Tumor organoid preparation method and device and application thereof
  • Tumor organoid preparation method and device and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0037] Example 1 Tumor organoid preparation and culture experiments

[0038] 1. Sample processing

[0039] 1. After spraying alcohol on the lung cancer tissue sample tube, put it into a biological safety cabinet.

[0040] 2. Mark the sample information on two 10cm Petri dishes respectively, and take out the tissue and put them into the Petri dishes.

[0041] 3. Add 10ml pre-cooled DPBS to each petri dish to wash the samples.

[0042] 4. After cleaning, add 10ml of lung cancer organoid medium to each dish, put a ruler at the bottom, and record the size of the sample with a camera.

[0043] 5. Take out the 1.5ml EP tube and cell cryopreservation tube, mark the sample information on the tube wall, and add RNAlater 500μl and 1ml 4% paraformaldehyde to the EP tube to preserve the DNA of cancerous tissue and normal tissue , RNA and tissue section raw materials.

[0044] 6. Cut the tissue into small pieces, and keep the materials used to detect the sample DNA, RNA and tissue sectio...

Embodiment 2

[0071] Embodiment 2 The tumor sample grinding and filtering integrated device of the present invention and its application

[0072] see in conjunction Figure 3 to Figure 6 As shown, according to an embodiment of the present invention, an integrated device for grinding and filtering tumor samples is provided, including a grinding hammer 1, a rubber plug 11 is constructed at the top of the grinding hammer 1, and the rubber plug at the top of the grinding hammer 1 is connected to the rubber plug. The sample holding filter cavity 2 is in sealing fit, and the grinding hammer 1 is freely separable from the sample holding filter cavity 2 . Specifically, the 100 μm filter 21 in the sample holding filter cavity 2 is used to intercept the tumor tissue sample, and the grinding hammer 1 is in full contact with the tumor tissue to grind the tissue. The culture medium in the sample holding filter cavity 2 is fully mixed with the ground tissue. The organoid holding filter chamber 4 is det...

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Abstract

The invention provides a tumor organoid preparation method and device and application thereof. The method comprises the following steps: obtaining a tumor tissue sample, cutting the tumor tissue sample into tissue blocks suitable for mechanical grinding; grinding the tissue blocks through a mechanical grinding method, performing first-stage filtration through a first-stage filter screen with a pore diameter of 80-200 [mu]m to form first-stage filtrate; and carrying out second-stage filtration on the first-stage filtrate by a second-stage filter screen with a pore diameter of 30-60 [mu]m to obtain a solid on the second-stage filter screen, thereby obtaining the tumor organoid. The tumor organoid is obtained by a simple method, the operation is easy and convenient, the survival rate and number of the organoid are remarkably increased, and the organoid culture time is greatly shortened.

Description

technical field [0001] The invention relates to the technical field of in vitro tumor models, in particular to a method for preparing tumor organoids and its device and application. Background technique [0002] Organoids are three-dimensional structures formed by self-organization of various types of cells derived from stem cells, which can simulate the structure and function of the organ where the cells are located. Compared with traditional 2D cell culture and animal models, organoids have incomparable advantages. The cell composition and function of organoids are very similar to in vivo tissues. Many organoids can maintain the stability of their genomes after long-term culture in vitro. This huge advantage enables them to more accurately reflect the effects of drugs. Compared with animal models, organoids can reduce the complexity of experiments and be used in real-time imaging techniques, which enables people to use this model to study the mechanism of human developmen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/12C12M1/00C12M3/00C12N5/09
CPCC12M45/02C12M33/14C12M21/08C12N5/0693C12N5/0697
Inventor 王宪宁宋凡
Owner 奥格诺德生物科技(北京)有限公司
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