3D co-culture model based on tumor cell and tumor-associated fibroblast

A fibroblast, tumor-related technology, applied in 3D co-culture model, new 3D co-culture model and its construction field, can solve the problem that tumor cells are difficult to be removed by chemotherapy drugs, and achieve the effect of fast three-dimensional structure and low cost

Inactive Publication Date: 2018-05-22
FUDAN UNIV
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Problems solved by technology

This mutual promotion of the two cells makes it dif...

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  • 3D co-culture model based on tumor cell and tumor-associated fibroblast
  • 3D co-culture model based on tumor cell and tumor-associated fibroblast
  • 3D co-culture model based on tumor cell and tumor-associated fibroblast

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Embodiment 1

[0025] 1. Materials

[0026] Human pancreatic cancer cell line Capan-1:

[0027] Human primary pancreatic stellate cells: derived from pancreatic cancer specimens isolated from Zhongshan Hospital affiliated to Fudan University.

[0028] DMEM medium: purchased from Thermo Company, USA,

[0029] DMEM / F12 medium: purchased from Hyclone Company of the United States,

[0030] 10% fetal bovine serum: purchased from Gibco, USA,

[0031] Trypsin:

[0032] PBS: Phosphate buffered saline (1×), 0.0067M (PO4)

[0033] Methylcellulose: purchased from Sigma, USA,

[0034] Untreated 96-well round bottom plate: purchased from Thermo Company, USA,

[0035] GFP:

[0036] RFPs:

[0037] Streptomycin-penicillin solution: purchased from Thermo Company, USA.

[0038] 2. Method

[0039] (1) Culture of primary pancreatic stellate cells

[0040] Quickly place the pancreatic cancer tissue obtained from the hospital in a 10 cm culture dish, cut it into small pieces with high-pressure steam st...

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Abstract

The invention belongs to the field of biomedicine, and relates to a 3D co-culture model based on a tumor cell and a tumor-associated fibroblast. The tumor cell is labeled with a green fluorescent protein, the tumor-associated fibroblast is labeled with a red fluorescent protein, and the two cells and a prepared methylcellulose gel medium are used to construct the 3D co-culture model. The 3D modelprepared in the invention simulates the anaerobic environment of in-vivo tumors, and the tumor cell and the tumor-associated fibroblast are co-cultured to construct the mutual promotion effect of thetwo cells in the in-vivo tumor microenvironment. The 3D co-culture model formed by the tumor cell and the tumor-associated fibroblast simulates the anaerobic environment of the in-vivo tumor, providesa similar effect to the in-vivo tumor microenvironment, and has application prospects in basic researches and clinical drugs.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a novel 3D co-culture model and a construction method thereof, in particular to a 3D co-culture model based on tumor cells and tumor-related fibroblasts. The 3D co-culture model can be used for basic research and establishment In vitro drug screening model. Background technique [0002] According to reports, the gradual increase in the incidence and mortality of cancer has become the main cause of death among Chinese people, seriously threatening the health of Chinese people. In recent years, with the gradual improvement of surgical techniques and the application of standardized and standardized radiotherapy and chemotherapy, the clinical treatment level of cancer has made great progress, but the five-year survival rate of cancers such as pancreatic cancer and non-small cell lung cancer is not as good as not effectively improved. The reason is mainly because the efficacy of clinical ch...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12N5/071C12Q1/02
CPCC12N5/0693C12N2500/84C12N2501/90C12N2502/22C12N2503/02C12N2513/00C12N2533/78G01N33/5011G01N2500/10
Inventor 党永军杨宸李增霞倪晓凌崔照盟高强王嘉琦
Owner FUDAN UNIV
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