Preparation method and application method of kit for fast testing efficacy of antitumor drug

An anti-tumor drug and kit technology, applied in the field of environmental science, can solve the problem that the PDX model cannot be used with immunosuppressants, the evaluation of the efficacy of immunomodulators, the time-consuming drug susceptibility test in the model building cycle, and the tumor formation rate of PDX model transplantation. Low problems, to achieve the effect of rapid and efficient detection, suitable for clinical use, and easy to promote

Active Publication Date: 2019-10-25
NANTONG UNIVERSITY
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AI Technical Summary

Problems solved by technology

[0004] The present invention provides a method for preparing and using a rapid drug efficacy test kit for anti-tumor drugs, which solves the problems in the prior art, such as the existence of certain errors, the long time spent on model building and drug sensitivity testing, and the low tumor formation rate of PDX model transplantation and other technical issues
Since none of the modeled mice has a mature immune system, the PDX model cannot be used to evaluate the efficacy of immunosuppressants and immunomodulators

Method used

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  • Preparation method and application method of kit for fast testing efficacy of antitumor drug
  • Preparation method and application method of kit for fast testing efficacy of antitumor drug
  • Preparation method and application method of kit for fast testing efficacy of antitumor drug

Examples

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Effect test

Embodiment 1

[0046] Such as Figure 1-4 Shown, a kind of preparation method of antineoplastic drug rapid efficacy test kit, the steps are:

[0047] Step 1: Take freshly collected clinical tumor samples or PDX mouse tumor samples in transport preservation tubes, which are filled with liquid nitrogen, and transported to the laboratory for subsequent operations in the shortest possible time.

[0048] The second step, preparation of single cell suspension: remove non-tumor tissue and necrotic tissue from clinical tumor samples or PDX mouse tumor samples in the laboratory, and cut tumor samples into 2mm 3 For small pieces, wash with PBS and collect the tissue pieces, centrifuge at 1000rpm for 5min to remove the supernatant; use type 2 collagenase to incubate the precipitate at 37°C for 2 to 4 hours on a shaker; filter with a 40μm nylon mesh after incubation to remove tissue residues Blocks and cell clusters; centrifuge the filtrate at 1000rpm for 5min to remove the supernatant, resuspend the p...

Embodiment 2

[0068] Such as Figure 1-4 Shown, a kind of preparation method of antineoplastic drug rapid efficacy test kit, the steps are:

[0069] Step 1: Take freshly collected clinical tumor samples or PDX mouse tumor samples in transport preservation tubes, which are filled with liquid nitrogen, and transported to the laboratory for subsequent operations in the shortest possible time.

[0070] The second step, preparation of single cell suspension: remove non-tumor tissue and necrotic tissue from clinical tumor samples or PDX mouse tumor samples in the laboratory, and cut tumor samples into 2mm 3 For small pieces, wash with PBS and collect the tissue pieces, centrifuge at 1000rpm for 5min to remove the supernatant; use type 2 collagenase to incubate the precipitate at 37°C for 2 to 4 hours on a shaker; filter with a 40μm nylon mesh after incubation to remove tissue residues Blocks and cell clusters; centrifuge the filtrate at 1000rpm for 5min to remove the supernatant, resuspend the p...

Embodiment 3

[0091] Such as Figure 1-4 Shown, a kind of preparation method of antineoplastic drug rapid efficacy test kit, the steps are:

[0092] Step 1: Take freshly collected clinical tumor samples or PDX mouse tumor samples in transport preservation tubes, which are filled with liquid nitrogen, and transported to the laboratory for subsequent operations in the shortest possible time.

[0093] The second step, preparation of single cell suspension: remove non-tumor tissue and necrotic tissue from clinical tumor samples or PDX mouse tumor samples in the laboratory, and cut tumor samples into 2mm 3 For small pieces, wash with PBS and collect the tissue pieces, centrifuge at 1000rpm for 5min to remove the supernatant; use type 2 collagenase to incubate the precipitate at 37°C for 2 to 4 hours on a shaker; filter with a 40μm nylon mesh after incubation to remove tissue residues Blocks and cell clusters; centrifuge the filtrate at 1000rpm for 5min to remove the supernatant, resuspend the p...

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Abstract

The invention discloses a preparation method and application method of a kit for fast testing the efficacy of an antitumor drug. The preparation method and application method includes: collecting a clinical tumor surgery sample or a PDX mouse tumor sample, preparing a single-cell suspension, extracting tumor stem cells and macrophages, building a 3D co-culture device, using LDH to detect tumor stem cell activity, and calculating the tumor stem cell activity increasing percentage of an administration group according to the tumor stem cell activity of the control to evaluate the efficacy of thecorresponding drug. Due to the fact that the tumor stem cells are hard to grow in an in-vitro manner, the method screens out tumor stem cells with high multiplication capacity first, tumor is formed by the aid of the tumor stem cells, and the efficacy result can be obtained in two weeks. The efficacy of the antitumor drug can be fast evaluated by combining in-vivo and in-vitro experiment technologies, and the efficacy of the antitumor drug can be fast and efficiently tested by the kit.

Description

technical field [0001] The invention relates to the field of environmental science and technology, in particular to a preparation method and use method of a rapid drug efficacy test kit for antineoplastic drugs. Background technique [0002] The fatality rate of tumor ranks first among all diseases, and it continues to rise, becoming a great enemy that threatens people's health. Half of all people will develop cancer at some point in their lives. Chemotherapy is currently the most common cancer treatment, which can effectively kill cancer cells. However, for some patients, not only do they not respond to chemotherapy drugs, but their healthy tissues are also destroyed, resulting in unnecessary toxic side effects for patients. Therefore, the treatment of tumors must be developed towards individualized and precise treatment. A major challenge in the research process is to develop powerful chemotherapy drug evaluation models for clinical patients. These models will replace pa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02C12N5/095C12N5/0786
CPCG01N33/5011C12N5/0695C12N2503/02G01N2500/10C12N2502/1157C12N2513/00
Inventor 王国华周佳敏高纯一张潇逸吴昊杨宇轩王祎晨徐丽华
Owner NANTONG UNIVERSITY
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