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Tumor cell three dimensional culture method based on folded non-woven polyester fiber strips

A non-woven polyester fiber, three-dimensional culture technology, applied in the direction of tumor/cancer cell, cell culture support/coating, 3D culture, etc. Aggregation, the effect of facilitating reproduction and expansion

Active Publication Date: 2020-06-05
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 (tumor cell three-dimensional culture scaffold, blood vessels and the interaction between cells. Chinese Tissue Engineering Research, 2013,17(42):7442-7448.)

Method used

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  • Tumor cell three dimensional culture method based on folded non-woven polyester fiber strips
  • Tumor cell three dimensional culture method based on folded non-woven polyester fiber strips
  • Tumor cell three dimensional culture method based on folded non-woven polyester fiber strips

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Preparation of scaffold for three-dimensional culture of tumor cells

[0056] 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.

[0057] The length of the short side of the obtained support 1 is 0.6cm; the total length of the long side is 3.6cm, and the long side is in a zigzag shape, the length of each line segment is 1.2cm, 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 inters...

Embodiment 2

[0068] The three-dimensional culture method of embodiment 2 tumor

[0069] 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.

[0070] 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

[0071] Example 3 Tumor three-dimensional culture method

[0072] In a 50ml plastic tube with an inner diameter of 3cm, put 1g of scaffold 1 in total, 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.

[0073] Insert human lung cancer cell A549 1×10 6 cells, 37°C, 5% CO 2 cultured under conditions such as Figure 5 shown.

[0074] Application Example 1 Expansion of A549 Cells

[0075] In a 50ml plastic tube with an inner diameter of 3cm, put 1g of scaffold 1, wash with PBS for 3 times, add PBS, sterilize at 121°C for 15min, discard the PBS after sterilization, and add 25ml of medium 1640+10% fetal bovine serum.

[0076] Access to A549 cells 1×10 6 pcs, 37°C, 5% CO 2 Conditioned culture, continuous culture for 6 days, counted by cell counting plate.

[0077] Replace scaffold 1 with scaffolds 2-5 prepared in the comparative example, and use them for three-dimensional...

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Abstract

The invention provides a tumor cell three dimensional culture method based on folded non-woven polyester fiber strips. The tumor cell three dimensional culture method comprises the specific steps of placing a bracket in a culture dish, adding a phosphoric acid buffer salt solution, washing the bracket, then performing sterilization, and adding a culture medium; performing inoculating with cells tobe tested for culturing, wherein the bracket is the non-woven polyester fiber strips which are folded through zigzag folding twice, the length ratio of the short edges to the long edges of the stripsis 1 to (4.5 to 6), the long edges are Z-shaped folded lines, folding lines formed by folding are parallel to the short edges of the non-woven polyester fiber strips and are intersected with the toppoints of the Z-shaped folded lines, and the surface inclined angle of folded surfaces formed by folding is 165-170 degrees. The method is simple to make, a good space structure can be formed, and cell propagation and amplification are facilitated. In vivo tumor microenvironment can be simulated. The invention further provides an application of the three dimensional culture method of the tumor cells, to drug screening, and the tumor tissue growth microenvironment basically consistent with the in vivo microenvironment can be simulated.

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 Applications(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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