Tumor in-vitro culture method and clinical chemotherapeutic drug screening method

An in vitro culture and culture method technology, applied in the direction of tumor/cancer cells, animal cells, pharmaceutical formulations, etc., can solve the problems of low tumor formation rate, death, and long model building cycle of PDX model transplantation, so as to speed up drug sensitivity Experimental progress, convenient operation, and the effect of shortening the time of tumor formation

Pending Publication Date: 2019-05-14
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the main difficulty of using PDX models for individualized precision medicine lies in the long time spent on model building and drug sensitivity testing, as well as the low tumor formation rate of PDX models.
For patients with rapid tumor development and short survival period, some patients died without drug sensitivity results

Method used

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  • Tumor in-vitro culture method and clinical chemotherapeutic drug screening method
  • Tumor in-vitro culture method and clinical chemotherapeutic drug screening method
  • Tumor in-vitro culture method and clinical chemotherapeutic drug screening method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0048] S301: Collect a tumor tissue sample.

[0049] In this embodiment, the tumor tissue sample is a breast tumor tissue from an operation sample of a clinical patient.

[0050] S302: Process the tumor tissue sample, and sort out tumor single cells and macrophages.

[0051] In this embodiment, the specific steps for processing tumor tissue samples include:

[0052] Exclude non-tumor tissue and necrotic tissue, and cut tumor tissue samples into 2mm 3 of small pieces. Wash with PBS buffer and collect the tissue pieces, then centrifuge at 1000rpm for 5min to remove the supernatant. Add type II collagenase to the tissue block and incubate at 37°C for 4h on a shaker. Filter through a 40 μm nylon mesh to remove tissue residues and cell clusters to obtain the first cell suspension, and centrifuge at 1000 rpm for 5 min to remove the supernatant to obtain the first cell pellet. The first cell pellet was resuspended in PBS buffer containing 1% fetal bovine serum (FBS) and washed, ...

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Abstract

The invention relates to the field of drug efficacy screening, and particularly discloses a tumor in-vitro culture method. The tumor in-vitro culture method includes the steps that tumor tissue samples are collected; the tumor tissue samples are treated, and single tumor cells and macrophages are sorted; the single tumor cells are placed in an environment full of hydrogel and then are screened toobtain tumor stem cells; the tumor stem cells and the macrophages are adopted to build a co-culture model to obtain in-vitro culture tumor samples. In this way, the tumor stem cells with high proliferation and differentiation capabilities can be screened out in vitro, therefore, the tumor stem cells are implanted into the animal body, the tumor formation time can be greatly shortened, the tumor formation rate is increased, and then susceptibility testing is conducted; therefore, the progress of susceptibility testing is speeded up, and time for treating patients is bought.

Description

technical field [0001] The invention relates to the field of drug efficacy screening, in particular to a method for culturing tumors in vitro and a method for screening clinical chemotherapy 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. However, there are many problems in the existing cancer treatment methods, the uncertainty of drug efficacy, drug resistance and severe side effects are very likely to occur during long-term use; in many cases, the more effective treatment method is often a combination of multiple drugs. But this is more likely to produce unknown drug reactions, and it is impossible to find the optimal treatment method in a short time. Therefore, the treatment of tumors must be developed towards individualized and precise treatment. Human-derived tumor xenograft (Patient-Derived Xenograft, PDX) model is to directly transplant th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/095A61K49/00
Inventor 王国华高纯一周佳敏王祎晨杨宇轩
Owner NANTONG UNIVERSITY
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