Three-dimensional cell culture medium for anti-tumor drug screening system and preparation of three-dimensional cell culture medium

A three-dimensional culture, tumor cell technology, applied in the direction of tumor/cancer cells, animal cells, vertebrate cells, etc., can solve the problem of unfavorable three-dimensional cultured cells in animal experiments xenotransplantation, the promotion and application of unfavorable cell three-dimensional culture technology, and the high experimental cost. and other problems, to achieve a good three-dimensional cell culture effect, easy subsequent analysis, and no toxic side effects.

Inactive Publication Date: 2016-11-23
GUANGDONG UNIV OF TECH
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  • Summary
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, Matrigel, a relatively mature and commonly used three-dimensional cell culture substrate, is expensive and has poor mechanical properties, which will cause high experimental costs and is not conducive to xenotransplantation of three-dimensional cultured cell animal experiments, which is not conducive to the promotion of three-dimensional cell culture technology. application

Method used

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  • Three-dimensional cell culture medium for anti-tumor drug screening system and preparation of three-dimensional cell culture medium
  • Three-dimensional cell culture medium for anti-tumor drug screening system and preparation of three-dimensional cell culture medium
  • Three-dimensional cell culture medium for anti-tumor drug screening system and preparation of three-dimensional cell culture medium

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Matrigel, sodium hyaluronate, and sodium alginate were mixed in appropriate proportions to culture three-dimensional cell culture substrates for prostate cancer PC3 cells, and then H.E. staining was performed to observe the morphology of three-dimensional cell culture. Specific steps are as follows:

[0038] (1) Mix matrigel, 10mg / ml sodium hyaluronate solution and 2% (W / V) medium-viscosity sodium alginate solution in an appropriate mixing ratio, and finally make the content of matrigel 10%, hyaluronic acid The final concentration of sodium alginate was 2.5 mg / ml, and the final concentration of sodium alginate was 0.5% (W / V).

[0039] (2) Place the mixture obtained in the above (1) on ice for 10 minutes to eliminate air bubbles, and then wrap PC3 cells to make 1.0x10 6 cells / ml mixture.

[0040] (3) Drop the mixed solution containing cells obtained in (2) into 0.1M CaCl 2 In the solution, let it stand for 10-20min to form milky white sodium alginate beads.

[0041] ...

Embodiment 2

[0049] Matrigel, sodium hyaluronate, and sodium alginate were mixed in appropriate proportions to culture three-dimensional cell culture substrates to culture prostate cancer DU145 cells, and then H.E. staining was performed to observe the morphology of three-dimensional cell culture. Specific steps are as follows:

[0050] (1) Mix matrigel, 10mg / ml sodium hyaluronate solution and 2% (W / V) medium-viscosity sodium alginate solution in an appropriate mixing ratio, and finally make the content of matrigel 10%, hyaluronic acid The final concentration of sodium alginate was 2.5 mg / ml, and the final concentration of sodium alginate was 0.5% (W / V).

[0051] (2) Place the mixture obtained above on ice for 10 minutes to eliminate air bubbles, and then wrap PC3 cells to make 1.0x10 6 cells / ml mixture.

[0052] (3) Drop the mixture containing cells obtained above into 0.1M CaCl 2 In the solution, let it stand for 10-20min to form milky white sodium alginate beads.

[0053] (4) The so...

Embodiment 3

[0062] Immunofluorescence staining experiment to observe the expression of N-Cadherin and Vimentin

[0063] Figure 5 It is the N-Cadherin immunofluorescence staining picture of the paraffin section after PC3 cells are cultured in the three-dimensional culture matrix of the present invention for 14 days, and Figure 5-1 and Figure 5-2 They are the pictures of N-Cadherin immunofluorescent staining without nuclear staining and the pictures of only nuclear staining without N-Cadherin immunofluorescent labeling of PC3 cells cultured in the three-dimensional culture substrate of the present invention for 14 days.

[0064] Image 6 It is the N-Cadherin immunofluorescence staining picture of PC3 cells in two-dimensional culture, and Figure 6-1 and Figure 6-2 They are the pictures of N-Cadherin immunofluorescence staining without nuclear staining and the picture of only nuclear staining without N-Cadherin immunofluorescence labeling of PC3 cells in two-dimensional culture.

[...

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Abstract

The invention discloses a novel medium for three-dimensional tumor cell culture. The three-dimensional cell culture medium can be used for wrapping tumor cells for three-dimensional culture so that anti-tumor drugs can be further screened. According to preparation of the three-dimensional cell culture medium, a sodium alginate solution, a sodium hyaluronate solution and an extracellular culture medium Matrigel are mixed in a proper proportion, and then cells are wrapped. Sodium alginate can be used for directly wrapping cells, but is bad for proliferation and differentiation of tumor cells, while Matrigel is an ideal cell three-dimensional culture medium bu is high in price, poor in mechanical strength and bad for screening and pharmacological research of anti-tumor drugs. The mechanical property of the medium is improved; meanwhile, formation of the tissue structure of extracellular culture cells is effectively promoted, usage of Matrigel is reduced or replaced, the cost of tumor cell three-dimensional culture is greatly reduced, xenograft and animal experiments can be conveniently carried out on three-dimensional cells, and the preparation method is simple and high in practicability.

Description

technical field [0001] The invention relates to a biological material for three-dimensional culture of tumor cells, in particular to a three-dimensional cell culture substrate composed of sodium alginate, sodium hyaluronate and Matrigel at an appropriate concentration and ratio. Background technique [0002] Cancer is a malignant disease that seriously threatens human health. The World Health Organization estimates that 7.6 million cancer patients died in the world in 2008, and 1.96 million people died in China, accounting for 25.89% of the global cancer deaths. The data of the third national retrospective sampling survey on causes of death show that with the development of my country's health services and the continuous improvement of medical technology, certain diseases such as acute infectious diseases and maternal and child diseases have been effectively controlled, and the mortality rate of malignant tumors However, it shows an obvious upward trend and has become one of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12Q1/02
Inventor 杜志云董宇琴郑希张秋炎王华倩张焜
Owner GUANGDONG UNIV OF TECH
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