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A three-dimensional culture method of tumor stem cells

A technology of three-dimensional culture of tumor stem cells, which is 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 convenient use

Active Publication Date: 2022-07-19
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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  • A three-dimensional culture method of tumor stem cells
  • A three-dimensional culture method of tumor stem cells
  • A three-dimensional culture method of tumor stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0033] 2. Add 0.4800g of polycaprolactone to 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 to stir for 12h, and then add 0.096g of fibroin , stir evenly to obtain spinning solution. The concentration of polycaprolactone in the spinning solution is 0.15 g / mL, and the mass ratio of polycaprolactone to JK1 and fibroin is 1:0.1:0.2.

[0034] 3. Use a glass syringe to suck up 1mL of the spinning solution in step 2, and connect a stainless steel needle to the glass syringe filled with the spinning solution, an...

Embodiment 2

[0038]In step 2 of this example, 0.4800 g of polycaprolactone was added to 3.2 mL of a mixed solution with a volume ratio of chloroform and methanol of 7:1, stirred at room temperature for 12 h, and then 0.00480 g of JK1 was added, continued to stir for 12 h, and then 0.096 g of fibroin was added, and the mixture was stirred evenly to obtain a spinning solution. The concentration of polycaprolactone in the spinning solution is 0.15 g / 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 in 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 the same as in Example 1.

Embodiment 3

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

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Abstract

The invention discloses a three-dimensional culture method of tumor stem cells. A honeycomb-shaped three-dimensional culture scaffold is 3D printed by using a photosensitive resin as a raw material. 2 The polycaprolactone of the S sustained-release donor JK1 was electrospun at the bottom of the culture scaffold, and the surface of the electrospun membrane was modified with growth factors to prepare a three-dimensional culture scaffold, which was then used to culture tumor stem cells. In the present invention, photosensitive resin is selected as the material of the 3D printing bracket, which has good biocompatibility and high mechanical strength; 2 The S donor JK1 is incorporated into electrospinning, and the pH of the medium can be changed by changing the pH of the medium. 2 The release of S is regulated to regulate the growth rate of cells; the growth factor is modified on the electrospinning surface to promote the proliferation of cancer stem cells. The invention utilizes the membrane-covered honeycomb culture scaffold to culture tumor stem cells with high quality and high efficiency, effectively promotes the proliferation of tumor stem cells, maintains their stemness and avoids differentiation, and lays a foundation for the characteristic research of tumor stem cells and the 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 of a novel tumor stem cell. Background technique [0002] 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. They are capable of self-renewal, multidirectional differentiation and unlimited proliferation. Expressed with stem cell surface markers. Studies have shown that CSCs have the ability to regenerate tumor cells and contribute to tumor heterogeneity, multidrug resistance, radiation resistance, and 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 for improving cancer cure rates and reducing tumor recurrence rates. However, because the number of CSCs in th...

Claims

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Application Information

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