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Single-layer or multi-layer 3D brain glioma cell culture model, and construction method and application thereof

A glioma, cell culture technology, applied in 3D culture, tumor/cancer cells, animal cells, etc., can solve the problem that plane culture is difficult to simulate, achieve less nucleic acid residue, ensure homogeneity, and remove cells full effect

Active Publication Date: 2019-09-20
BEIJING TIANTAN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Tumor cells interact with these factors to form corresponding biological behaviors through direct contact, paracrine, etc. However, it is difficult to simulate these factors in the existing 2-dimensional plane culture, and it is also difficult to simulate these factors in plane culture. Representative Tumor Models

Method used

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  • Single-layer or multi-layer 3D brain glioma cell culture model, and construction method and application thereof
  • Single-layer or multi-layer 3D brain glioma cell culture model, and construction method and application thereof
  • Single-layer or multi-layer 3D brain glioma cell culture model, and construction method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0056] Example 1 Construction of single-layer and multi-layer 3D glioma cell models

[0057] (1) Preparation of sterile slices of SD rat decellularized brain

[0058] 32-week-old female SD rats were perfused with 200ml of normal saline in the abdominal aorta for 5 minutes until the brain tissue turned white, and then the brain was removed by craniotomy; the obtained SD rat brain was removed from the cerebellum and placed at -80°C. 30min, and then placed at room temperature for 30min, repeated four times of alternate freezing and thawing; the brain after alternate freezing and thawing was treated with 100ml deionized water at room temperature for 4h at 60rpm. Then treated with 50 ml of 1% Triton-X100 at 4°C for 12 hours at 60 rpm to obtain preliminary decellularized brain tissue. Then wash with deionized water at room temperature for 3 times at 60 rpm, 10 min / time. Then at 60 rpm, 50 ml of 4% sodium deoxycholate was treated at room temperature for 12 hours to obtain decellula...

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Abstract

The invention discloses a single-layer or multi-layer 3D brain glioma cell culture model, and a construction method and an application thereof. The construction method includes the steps: freezing a decellularized brain tissue, and cut into slices; freeze-drying the slices, and sterilizing to obtain a scaffold; and inoculating a single-slice or multi-slice slice scaffold with brain glioma cells to obtain a single-layer or multi-layer brain glioma cell 3D culture model. The used decellularized brain slice scaffold for constructing the 3D brain glioma model has the advantages of complete removal of cells and fewer nucleic acid residues, can partially preserve a stereoreticular structure and biological components of an original brain matrix, is suitable for growth of the brain glioma cells, and can simulate a microenvironment of the growth of the brain glioma cells in vivo; the multi-layer 3D brain glioma cell culture model can also simulate the hypoxia and nutrition gradient of glioma in vivo, is more close to the brain glioma cells growing in vivo, and is suitable for screening of antitumor drugs, co-culture of the brain glioma cells and tumor-related cells, and study on the mechanism of action between the brain glioma cells and ECM.

Description

technical field [0001] The present invention relates to a cell culture model, in particular to a single-layer or multi-layer 3D glioma cell culture model and its construction method. The present invention also relates to the use of the glioma cell model in experimental research on glioma or anti-tumor drugs The application in screening belongs to the field of brain glioma cell culture model construction and application. Background technique [0002] Glioma is the most common primary malignant tumor of the central nervous system, with an annual incidence of 4 / 10 to 5 / 100,000. With the development of cell biology and molecular pathology, it has been confirmed that glioma is a kind of tumor with high heterogeneity. The molecular phenotypes of glioma cells in the same part of the same patient are very different, and the molecular Phenotype is closely related to treatment response. In the current experimental research, glioma cell lines (such as U87, LN229, U251, etc.) or prima...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02
CPCC12N5/0693C12N2513/00G01N33/5011G01N2500/10
Inventor 李文斌崔磊王灿李晓玉
Owner BEIJING TIANTAN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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