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Extracorporal building method of tumor microenvironment and application of method to drug allergy screening

A technology of tumor microenvironment and construction method, applied in the field of in vitro construction of tumor microenvironment, can solve the problems of toxicity, strong cells, and high price of animal gels

Inactive Publication Date: 2015-02-11
SUZHOU CANCERCELL BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the commonly used matrices are derived from animal gels such as collagen I, gelatin or extracted extracellular matrix, but animal gels are expensive and contain many uncertain components
[0006] Chinese patent CN102952279A has announced the method for preparing hydrogel with methyl vinyl ether / maleic acid copolymer and cross-linking agent reaction and its application in tumor tissue culture, but because contain a large amount of cell growth factors (such as epidermal growth factor, fibroblast growth factor, and tissue plasminogen activator), uncertainty about the effects of these cytokines on cultured cells
The disadvantage is that the main material used, acrylamide, is easily decomposed by high heat to release irritating gas; and the acrylic acid transformed into an acid-base environment has a strong toxic effect on cells.

Method used

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  • Extracorporal building method of tumor microenvironment and application of method to drug allergy screening
  • Extracorporal building method of tumor microenvironment and application of method to drug allergy screening
  • Extracorporal building method of tumor microenvironment and application of method to drug allergy screening

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Embodiment 1: Silk fibroin three-dimensional matrix construction method

[0087] (1) Prepare 1%~5% silk fibroin solution

[0088] 1) Cut the cocoon shell into 1cm 2 The pieces of the silkworm cocoons were submerged in 0.5% sodium carbonate solution and boiled for 2-3 times, at least 1 hour each time.

[0089] 2) Wash 2-3 times with natural water first, then wash twice with deionized water and then dry.

[0090] 3) Heat to boiling with 50% calcium chloride solution (or dissolve in 9M lithium bromide solution), add dry silk fibroin and stir to fully dissolve silk fibroin, cool to room temperature and filter with Buchner funnel.

[0091] 4) Put the filtrate into a dialysis bag and dialyze with deionized water for 3-5 days to prepare a silk fibroin solution.

[0092] 5) Pack it in a fresh-keeping bag, put it in a -20°C refrigerator (12 hours), then put it in a -80°C refrigerator (6 hours), and finally put it in a freeze dryer for at least 24 hours.

[0093] 6) Weigh 10 g ...

Embodiment 2

[0117] Example 2: Screening for colorectal cancer chemotherapeutic drug sensitivity

[0118] (1) Whole cell extraction and inoculation of colorectal cancer tumor tissue

[0119] 1) Take the tumor tissue within 30 minutes of isolation, soak it in saline containing double antibody for 5 minutes, wash the blood stains on the specimen, put it in DMEM medium and cut it into a paste repeatedly.

[0120] 2) Collect the tissue fluid, remove the supernatant, add tissue digestion solution, carry out enzymatic hydrolysis at 37°C for 1-2 hours, and mix the digestion solution once every 15 minutes.

[0121] 3) When the tissue is soft and flocculent, filter it through a 200-mesh sieve, and gently grind it with a glass syringe needle, while continuously washing it with DMEM medium to disperse the tissue into a cell suspension.

[0122] 4) Centrifuge in a low-speed centrifuge for 10 minutes (1500 r / min), remove the supernatant, and add DMEM medium with 20% FBS. Count cells and adjust cell c...

Embodiment 3

[0136] Example 3: Breast cancer chemotherapy drug sensitivity screening

[0137] (1) Whole cell extraction and inoculation of breast cancer tissue

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Abstract

The invention relates to an extracorporal building method of a tumor microenvironment and an application of the method. By using the method, a three-dimensional stent similar with a cell internal microenvironment is built and total cells of tumor tissues are inoculated to promote recovery of physiological activity of all cells, so that a tumor microenvironment is built for drug screening for individual-based tumor treatment.

Description

technical field [0001] The invention relates to a method for constructing a tumor microenvironment in vitro and its application. The method constructs a three-dimensional scaffold similar to the microenvironment in the body of a cell, and can promote the recovery of the physiological activity of each cell after inoculation of the whole tumor tissue cells, thereby establishing a tumor microenvironment. environment, drug screening for personalized cancer therapy. Background technique [0002] With the continuous advancement of modern medicine and the deepening of people's understanding of malignant tumors, the cure rate of tumors has continued to increase in the past two decades (Cancer statistics 2014, CA Cancer J Clin 2014; 64:9–29), but malignant tumors in In the 35-59 age group, it still ranks first as the cause of death. According to statistics, by 2050, there will be 300 million cancer patients in the world, of which 200 million will lose their lives. [0003] Although...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02
Inventor 王守立
Owner SUZHOU CANCERCELL BIOTECH
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