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Three-dimensional culture method of tumor stem cells

A technology of three-dimensional culture of tumor stem cells, applied in the field of three-dimensional culture of new tumor stem cells, can solve the problems of easy aggregation and differentiation, slow proliferation of tumor stem cells, etc., and achieve the effect of fine structure, conducive to spherical growth, and maintenance of cell stemness

Active Publication Date: 2020-03-24
SHAANXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to solve the problem of slow proliferation and easy aggregation and differentiation of tumor stem cells in in vitro culture, and to provide a new three-dimensional culture method for tumor stem cells

Method used

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  • Three-dimensional culture method of tumor stem cells
  • Three-dimensional culture method of tumor stem cells
  • Three-dimensional culture method of tumor stem cells

Examples

Experimental program
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Effect test

Embodiment 1

[0032] 1. Use FreeCAD design software to make a honeycomb bracket model, inject acrylic resin into an LCD light-curing 3D printer and print a honeycomb-shaped culture bracket. The curing wavelength is 405nm. The structure of a honeycomb-shaped culture bracket is as follows: figure 1 As shown in a, it consists of a honeycomb cell culture rack (left) and a fixed support (right). The honeycomb cell culture frame is circular with a diameter of 18 mm. The height of the honeycomb cell is 1 mm and the side length is 0.5 mm.

[0033] 2. Add 0.4800g of polycaprolactone into 3.2mL of a mixture of chloroform and methanol with a volume ratio of 7:1, stir at room temperature for 12h, then add 0.0480g of JK1, continue stirring for 12h, then add 0.096g of fibroin , and stir evenly to obtain a spinning solution. The concentration of polycaprolactone in the spinning solution is 0.15g / mL, and the mass ratio of polycaprolactone to JK1 and fibroin is 1:0.1:0.2.

[0034] 3. Use a glass syringe to...

Embodiment 2

[0038]In step 2 of this embodiment, 0.4800g of polycaprolactone was added to 3.2mL of a mixture of chloroform and methanol with a volume ratio of 7:1, stirred at room temperature for 12h, then 0.00480g of JK1 was added, and stirred for 12h, and then Add 0.096g fibroin, stir evenly to obtain spinning solution. The concentration of polycaprolactone in the spinning solution is 0.15g / mL, and the mass ratio of polycaprolactone to JK1 and fibroin is 1:0.01:0.2. After electrospinning using the spinning solution according to the method of Step 3 of Example 1, a polycaprolactone / JK1 composite fiber membrane was attached to the bottom of the honeycomb cell culture frame, which was recorded as PCL-JK1-1%. Other steps are identical with embodiment 1.

Embodiment 3

[0040] In step 2 of this example, 0.4800g of polycaprolactone was added to 3.2mL of a mixture of chloroform and methanol with a volume ratio of 7:1, stirred at room temperature for 12h, then 0.0240g of JK1 was added, continued to stir for 12h, and then Add 0.096g fibroin, stir evenly to obtain spinning solution. The concentration of polycaprolactone in the spinning solution is 0.15g / mL, and the mass ratio of polycaprolactone to JK1 and fibroin is 1:0.05:0.2. After electrospinning using the spinning solution according to the method of Step 3 of Example 1, a polycaprolactone / JK1 composite fiber membrane was attached to the bottom of the honeycomb cell culture frame, which was recorded as PCL-JK1-5%. Other steps are identical with embodiment 1.

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Abstract

The invention discloses a three-dimensional culture method of tumor stem cells. 3D printing is performed by using photosensitive resin as a raw material to obtain a honeycomb-shaped three-dimensionalculture scaffold; electrostatic spinning of polycaprolactone containing an H2S slow-release donor JK1 is carried out on bottom of the culture scaffold and the surface of an electrospun membrane is modified with a growth factor to prepare a three-dimensional culture scaffold; and then the scaffold is used for culturing tumor stem cells. According to the invention, the photosensitive resin is selected as the material of the 3D printing scaffold and the material is good in biocompatibility and high in mechanical strength; the pH regulation type H2S donor JK1 is doped into electrostatic spinning and the release of H2S can be regulated and controlled by changing the pH of a culture solution so as to regulate and control the growth speed of cells; the growth factor is modified on the surface ofthe electrospun yarn to promote proliferation of tumor stem cells. The High-quality and high-efficiency culturing of the tumor stem cells is carried out by using the coated honeycomb-shaped culture scaffold; proliferation of the tumor stem cells is effectively promoted; meanwhile, stemness of the tumor stem cells is kept and differentiation is avoided, so that a foundation is laid for characteristic research of the tumor stem cells and development of targeted drugs.

Description

technical field [0001] The invention belongs to the technical field of stem cell culture, and in particular relates to a three-dimensional culture method for novel tumor stem cells. Background technique [0002] Cancer stem cells (cancer stem cells, CSCs) are a small part of the tumor cell population. They have the ability to promote tumorigenesis, maintain tumor growth and maintain tumor heterogeneity, and are capable of self-renewal, multidirectional differentiation and unlimited proliferation. Expression of stem cell surface markers. Studies have shown that CSCs have the ability to regenerate tumor cells and contribute to tumor heterogeneity, multidrug resistance and radiation resistance, as well as early micrometastasis. Therefore, CSCs are closely related to tumor metastasis, recurrence and drug resistance, and research and development of drugs and therapies targeting CSCs are crucial to improving the cure rate of cancer and reducing the rate of tumor recurrence. Howe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/095
CPCC12N5/0695C12N2513/00C12N2533/30
Inventor 段新瑞汤薇
Owner SHAANXI NORMAL UNIV
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