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A three-dimensional culture method of tumor cells based on folded non-woven polyester fiber tape

A non-woven polyester fiber and tumor cell technology, which is applied in the direction of tumor/cancer cells, cell culture support/coating, animal cells, etc., can solve the problems of no consensus on three-dimensional culture in vitro, so as to facilitate proliferation and migration, and facilitate Growth and multiplication, making simple effects

Active Publication Date: 2021-09-28
TONGDE HOSPITAL OF ZHEJIANG PROVINCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, there is currently no consensus on which method is the best for three-dimensional culture in vitro. The key lies in the material scaffold, culture system and how to better simulate the internal environment of the tumor (the interaction between three-dimensional tumor cell culture scaffold, blood vessels and cells[J]. Chinese Tissue Engineering Research, 2013,17(42):7442-7448.)

Method used

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  • A three-dimensional culture method of tumor cells based on folded non-woven polyester fiber tape
  • A three-dimensional culture method of tumor cells based on folded non-woven polyester fiber tape
  • A three-dimensional culture method of tumor cells based on folded non-woven polyester fiber tape

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1, the preparation of the scaffold of tumor cell three-dimensional culture

[0054] Take non-woven polyester fiber material (polyethylene terephthalate, gram weight 100 grams / square meter), as attached figure 1 The cutting shown is a rectangle with side a length of 6cm and side b length of 3cm. According to the accordion folding method, the creases divide the area of ​​the rectangle into thirds, the creases are perpendicular to side a, and the acute angle formed by the dotted line and side b is 60° °, the distance between the two dotted lines is 0.6cm, cut along the dotted line to get the bracket 1.

[0055] The length of the short side of the obtained support 1 is 0.6cm; the total length of the long side is 6.9cm, the long side is zigzag, the length of each line segment is 2.3cm, and the line angle between adjacent line segments is 120 °. figure 2 As shown, the creases formed by folding (crease 1, crease 2) are parallel to the short side and intersect with...

Embodiment 2

[0066] Embodiment 2, three-dimensional culture method of tumor

[0067] Put 1.0g of scaffold 1 into a 50ml plastic tube with an inner diameter of 3cm, wash with PBS three times, add PBS, sterilize at 121°C for 15min, discard the PBS after sterilization, and add 20ml of medium 1640+10 % fetal bovine serum, as attached image 3 Shown is the three-dimensional state of the scaffold without cells seeded.

[0068] Insert mouse Lewis lung cancer cells 1×10 6 pcs, 37°C, 5% CO 2 cultivated under conditions. as attached Figure 4 Shown is the three-dimensional state of the scaffold 24 hours after seeding cells.

Embodiment 3

[0069] Embodiment 3, tumor three-dimensional culture method

[0070] Put 1g of scaffold 1 in a 50ml plastic tube with an inner diameter of 3cm, wash 3 times with PBS, sterilize at 121°C for 15min after adding PBS, discard the PBS after sterilization, add 30ml of medium 1640 +10% fetal bovine serum.

[0071] Insert human lung cancer cell A549 1×10 6 cells, 37°C, 5% CO 2 cultured under conditions such as Figure 5 shown.

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Abstract

The invention provides a method for three-dimensional culturing of tumor cells based on folded non-woven polyester fiber strips. The specific steps include placing a bracket in a culture dish, adding a phosphate buffered saline solution to wash the bracket and then sterilizing it, and adding a culture medium; The cells to be tested are cultured; the support is a non-woven polyester fiber strip that has been accordion-folded twice, the length ratio of the short side to the long side of the strip is 1:4.5-6, and the long side is a zigzag fold line; The crease formed by folding is parallel to the short side of the non-woven polyester fiber tape and intersects with the apex of the zigzag fold line; the angle between adjacent folded surfaces formed by folding is 165-170°. The method is simple to manufacture, can form a good spatial structure, and is beneficial to the reproduction and expansion of cells. And can simulate the tumor microenvironment in vivo. The present invention also provides the application of the above-mentioned three-dimensional culture method of tumor cells in drug screening, which can simulate the growth microenvironment of tumor tissue basically consistent with that in vivo.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a three-dimensional culture method for tumor cells based on folded non-woven polyester fiber strips. Background technique [0002] The study of tumor biology mostly adopts two-dimensional monolayer cell culture technology in vitro, while experimental animal models are mainly used in vivo. Two-dimensional cell culture usually consists of cells growing on a single-layer two-dimensional plastic plane. The cells have good extensibility, the culture technology is relatively mature, and it has the advantages of simple operation, economical, intuitive observation indicators, and controllable culture conditions. [0003] However, studies have shown that the tumor microenvironment in vivo plays an important role in the proliferation, differentiation, metastasis and drug resistance of tumor cells. In the two-dimensional cell culture system, tumor cells grow in a single layer on the surface of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09
CPCC12N5/0062C12N5/0693C12N2533/30
Inventor 陈观平李静应栩华
Owner TONGDE HOSPITAL OF ZHEJIANG PROVINCE
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