Individualized drug sensitivity detection method for tumor tissue homogenate substituted serum micro-tissue block

A technology of tumor tissue and detection method, which is applied in the field of biomedicine, can solve the problems of long time-consuming evaluation of anti-tumor drugs and poor data correlation, and achieve the effect of reliable detection results, simple operation, and low requirements for experimental conditions

Pending Publication Date: 2022-03-11
CENT HOSPITAL TAIAN CITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In order to solve the deficiencies of the prior art, the present invention aims to provide an in vitro tissue culture liquid instead of serum micro-tissue block individualized tissue block culture system for anti-tumor drug susceptibility detection, no need to obtain tissue samples through surgery, and will not harm the patient. It causes great trauma; and it can detect tumors at an early stage, which solves the problems of time-consuming screening of personalized tumor treatment options and evaluation of anti-tumor drugs, and poor data correlation, and preserves the heterogeneity of tumors and the original microenvironment to the greatest extent. Guaranteed the efficacy of tumor tissue

Method used

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  • Individualized drug sensitivity detection method for tumor tissue homogenate substituted serum micro-tissue block
  • Individualized drug sensitivity detection method for tumor tissue homogenate substituted serum micro-tissue block
  • Individualized drug sensitivity detection method for tumor tissue homogenate substituted serum micro-tissue block

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Embodiment 1

[0030] After the tumor-bearing mice were sacrificed by cervical dislocation, they were transferred to an ultra-clean bench; the mouse tumor tissue pieces were transferred to a petri dish, and washed 1-2 times with PBS; the cleaned tissue pieces were cut into large parts with high activity about 1mm 3 uniform small pieces; weigh the rest of the tissue pieces, transfer them to a sterilized glass homogenizer, add serum-free medium according to the ratio of tissue piece (g): serum-free medium (ml) equal to 1:5, and grind Homogenate; transfer the ground tissue homogenate to a centrifuge tube, centrifuge at 15,000 rpm for 5 minutes; transfer the supernatant to a new centrifuge tube after centrifugation, pass through a 0.22um filter, and collect the filtrate; separate the filtrate and serum-free medium Dilute according to different volume ratios 1:6 (15%), 1:9 (10%), 1:19 (5%) to obtain a culture medium containing a certain proportion of tissue homogenate; in a 96-well plate, add 3 ...

Embodiment 2

[0032] The tumor organoids obtained by the method described in Example 1 were used for drug sensitivity test, and the test drugs included vincristine, epirubicin, pirarubicin, docetaxel, 5-fluorouracil, cisplatin, cyclophosphamide, gemcitabine and oxaliplatin. After 3 days of administration, add 20 μL of 5 mg / ml MTT solution to each well, continue to incubate for 4 hours, terminate the culture, aspirate the medium, add 150 μL of DMSO to each well, shake for 10 minutes, select a wavelength of 490 nm, and measure it on an enzyme-linked immunosorbent assay instrument Absorbance of each well.

[0033] Inhibition rate of cell activity %=(absorbance value of control group−absorbance value of experimental group) / absorbance value of control group*100%. When the inhibition rate is greater than 30%, the drug is effective.

[0034] Depend on figure 2 It can be seen that, according to the tumor tissue homogenate of 15%, 10%, and 5% concentrations, 10% autologous serum and 10% fetal bov...

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Abstract

The invention provides a tumor tissue homogenate substituted serum micro-tissue block individualized drug sensitivity detection method, and belongs to the technical field of biomedicine.The method mainly comprises the following steps that an artificial basement membrane three-dimensional tissue culture low-viscosity culture dish and a culture plate are prepared, obtained fresh tumor tissue is processed into tumor micro-tissue blocks uniform in size, and the tumor micro-tissue blocks are subjected to drug sensitivity detection; and placing the tissue block in a low-adhesion plate containing a tissue fluid substituted serum micro-tissue block individualized culture medium for stable incubation, inoculating the stabilized tumor micro-tissue block into a culture plate, and carrying out drug treatment and drug sensitivity detection. According to the method, the problems of long time consumption and poor data correlation in tumor personalized treatment scheme screening and antitumor drug evaluation are solved, the heterogeneity and the original microenvironment of tumors are reserved to the greatest extent, and the efficiency of tumor tissues is ensured. The method is easy to operate and high in feasibility, and the optimal individualized treatment scheme of the patient can be rapidly and accurately obtained within three days.

Description

technical field [0001] The present invention relates to the technical field of biomedicine, in particular to an individualized in vitro tumor tissue culture system of tumor tissue liquid instead of serum micro-tissue blocks. Drug susceptibility testing data provide clinical basis for personalized and precise treatment of cancer patients. Background technique [0002] In recent years, the incidence of malignant tumors has continued to rise, and surgery combined with chemotherapy is still the main means of cancer treatment. However, due to inter-individual differences, the sensitivity of the same type of tumor to different chemotherapeutic drugs varies. In vitro chemotherapy drug sensitivity test can help oncologists understand the sensitivity of different patients to different chemotherapy drugs in advance, formulate individualized chemotherapy regimens, and realize precise drug use for cancer patients. Therefore, establishing a simple-to-operate and easy-to-evaluate detect...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02G01N21/31C12N5/09
CPCG01N33/5011G01N21/31C12N5/0693G01N2500/10C12N2513/00C12N2533/40C12N2503/02C12N2500/90
Inventor 薛敬伟倪庆宾刘富垒赵斌刘少霞
Owner CENT HOSPITAL TAIAN CITY
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