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Three-dimensional breast cancer organoid model as well as culture method and application thereof

A culture method and breast cancer technology, applied in the field of tumor immunology, can solve the problems of poor control of the placement position of cells in the stent, lack of vascular structure in the stent, hindering long-term culture and perfusion, etc., to improve the macroscopic shape retention ability and repeatability. Good, improve the effect of biocompatibility

Pending Publication Date: 2022-05-10
安徽骆华生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although porous scaffolds possess the mechanical and chemical properties of ECMs for 3D tumor cell culture, disadvantages include lack of vascularity in the fabricated scaffolds, impeding perfusion for long-term cultures, and poor control of cell placement within the scaffold

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] A method for culturing a three-dimensional breast cancer organoid model, specifically comprising the steps of:

[0033] (1) Culture of breast cancer tumor cells:

[0034] A) Treatment of breast cancer tissue derived from a patient that has been separated from the human body: crush and separate the fresh breast cancer sample tissue, and culture the isolated tumor cells for 3 days to obtain the cell culture supernatant and filter it, and use Cell purification by differential attachment method to obtain purified primary breast cancer cells;

[0035] B) Expansion culture of primary breast cancer cells in vitro: the primary breast cancer cells are expanded and cultured using RPMI-1640 medium containing 10% fetal bovine serum, and the cell culture supernatant after expansion culture is taken;

[0036] (2) Construction of porous tissue scaffolds:

[0037] A) The whole porcine lung tissue was frozen at -25°C for 3 hours. After being fixed and shaped, the area without obvious ...

Embodiment 2

[0045] A method for culturing a three-dimensional breast cancer organoid model, specifically comprising the steps of:

[0046] (1) Culture of breast cancer tumor cells:

[0047]A) Treatment of breast cancer tissue derived from a patient that has been separated from the human body: crush and separate the fresh breast cancer sample tissue, and culture the isolated tumor cells for 3 days to obtain the cell culture supernatant and filter it, and use Cell purification by differential attachment method to obtain purified primary breast cancer cells;

[0048] B) Expansion culture of the primary breast cancer cells in vitro: the primary breast cancer cells are expanded and cultured in DMEM medium containing 10% fetal bovine serum, and the cell culture supernatant after the expansion culture is taken;

[0049] (2) Construction of porous tissue scaffolds:

[0050] A) The whole porcine lung tissue was frozen at -25°C for 3 hours. After being fixed and shaped, the area without obvious b...

Embodiment 3

[0058] A method for culturing a three-dimensional breast cancer organoid model, specifically comprising the steps of:

[0059] (1) Culture of breast cancer tumor cells:

[0060] A) Treatment of breast cancer tissue derived from a patient that has been separated from the human body: crush and separate the fresh breast cancer sample tissue, and culture the isolated tumor cells for 3 days to obtain the cell culture supernatant and filter it, and use Cell purification by differential attachment method to obtain purified primary breast cancer cells;

[0061] B) Expansion culture of primary breast cancer cells in vitro: the primary breast cancer cells are expanded and cultured in DMEM / F-12 medium containing 10% fetal bovine serum, and the cell culture supernatant after expansion culture is taken;

[0062] (2) Construction of porous tissue scaffolds:

[0063] A) The whole porcine lung tissue was frozen at -25°C for 3 hours. After being fixed and shaped, the area without obvious bro...

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Abstract

The invention discloses a three-dimensional breast cancer organ-like model and a culture method and application thereof, pig-derived lung tissue is used as a scaffold for constructing a tumor model, physiological and histological characteristics of pig-derived tissue are used as construction blocks, multicellular tissue with vasculature is created, a porous tissue scaffold is constructed, and the three-dimensional breast cancer organ-like model is obtained. Meanwhile, a composite sol solution of gelatin and sodium alginate is utilized to soak the scaffold, then freeze drying is performed, and a protective layer is formed on the outer surface of the porous tissue scaffold, so that the mechanical property and the macroscopic shape retaining ability of the scaffold are improved, and the support requirement of long-term culture of tumor organs is met; hydrogel is mixed with cell culture supernate obtained after amplification culture, the biocompatibility with the stent is improved, meanwhile, the culture of in-vitro three-dimensional breast cancer organoid is facilitated by changing the components of the hydrogel and specifically adding cell growth factors for promoting tumor cell growth, and the preparation process is simple and good in repeatability.

Description

technical field [0001] The invention relates to the technical field of tumor immunology, in particular to a three-dimensional breast cancer organoid model and its culture method and application. Background technique [0002] Traditional in vitro drug sensitivity test models for tumors mainly include human-derived tumor cell lines and human-derived tumor xenograft models. Human tumor cell lines can quickly obtain drug sensitivity screening results, but the culture method affects the maintenance of tumor characteristics of primary patients, and the accuracy of drug sensitivity screening is relatively low in consistency with the patient's in vivo response; human tumor xenograft models can The characteristics of primary patient tumors can be used as living tumors for preservation and passage, providing very valuable research specimens for oncology research, but the success rate of this model is low, and the drug screening test takes too long and costs too much. The organoid mod...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12N5/071C12Q1/02
CPCC12N5/0693C12N5/0631G01N33/5044G01N33/5076G01N33/5011C12N2513/00C12N2509/00C12N2533/54C12N2533/74C12N2535/00C12N2503/02C12N2533/76C12N2501/10G01N2500/10
Inventor 苗春光
Owner 安徽骆华生物科技有限公司