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3D bioprinting in-vivo tumor model and building method thereof

A construction method, a three-dimensional biological technology, applied in the field of biotechnology or three-dimensional bioprinting, can solve the problems of low tumorigenic rate, inability to simulate three-dimensional structures, affecting tumor diagnosis and treatment, and achieve high tumorigenic rate

Active Publication Date: 2020-06-19
THE FIRST AFFILIATED HOSPITAL OF WANNAN MEDICAL COLLEGE YIJISHAN HOSPITAL OF WANNAN MEDICAL COLLEGE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The tumor tissue block transplantation method is to cut fresh tumor tissue specimens into small tissue blocks or make homogenate and transplant them into nude mice. This method retains the original tumor tissue structure and the three-dimensional microenvironment of tumor cells, but due to the presence of The disadvantages such as high requirements for fresh tumor tissue parts, strict sampling time, and low tumorigenic rate limit its application.
[0004] The cell suspension injection method is to take the tumor cells in the logarithmic growth phase cultured in vitro, adjust to the appropriate cell concentration with serum-free culture medium or saline, and then inject them into nude mice. This method is easy to operate and has a high tumorigenic rate. However, since the tumor cells used for transplantation need to be digested with enzymes, the original cell structure of the tumor may be destroyed, and the two-dimensionally cultured tumor cells grow in a single layer in vitro, lacking the interaction between cells-cells and cell-extracellular matrix. It cannot simulate the three-dimensional structure of tumor tissue microenvironment in vivo, so it is different from tumor cells in vivo in terms of protein expression, cell signal transduction, cell activity and response to drugs.
These factors can lead to changes in the biological behavior of transplanted tumors, and ultimately affect the diagnosis and treatment of tumors

Method used

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  • 3D bioprinting in-vivo tumor model and building method thereof
  • 3D bioprinting in-vivo tumor model and building method thereof
  • 3D bioprinting in-vivo tumor model and building method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0150] Example 1: Preparation of in vivo tumor model by human glioma cell U251

[0151] 1. Human glioma cells U251 were cultured in DMEM high-glucose medium containing 10% fetal bovine serum, and the medium was replaced every 2-3 days. When the cells were in the logarithmic growth phase, 0.25% trypsin was routinely used Digest and collect by centrifugation for later use.

[0152] 2. Prepare solutions with mass volume fractions of 3% sodium alginate and 10% gelatin, and sterilize under high temperature and high pressure. First place the 3×10 6 / mL of glioma cells were resuspended in 1mL DMEM high-glucose medium, and then mixed with 4% sodium alginate and 20% gelatin solution at a volume ratio of 1:1:2 to obtain a final concentration of 5% gelatin, 0.75 % sodium alginate and 7.5 x 10 5 / mL mixed printing material of glioma cells.

[0153] 3. Through the present invention figure 1 The schematic diagram showing the principle and steps of 3D bioprinting and crosslinking for 3...

Embodiment 2

[0161] Example 2: Preparation of in vivo tumor model by human glioma cell U118

[0162]Specific experimental steps:

[0163] Human glioma cell line U118 cells were cultured in DMEM high-glucose medium containing 10% fetal bovine serum, and the medium was replaced every 2-3 days. When the cells were in the logarithmic growth phase, they were routinely centrifuged and collected for later use.

[0164] Sodium alginate and 20% gelatin solutions with mass volume fractions of 4% and 20% respectively were prepared and sterilized under high temperature and high pressure. First place the 4×10 6 / mL U118 cells were resuspended in 1mL cell culture medium, and then mixed with 4% sodium alginate and 20% gelatin solution at a volume ratio of 1:1:2 to obtain a final concentration of 10% gelatin, 1% sodium alginate and 1×10 6 / mL mixture of U118 cells.

[0165] A 3D multi-nozzle bioprinter was used to print a cube grid scaffold with a side length of 15 mm and a thickness of 1 mm, with a f...

Embodiment 3

[0176] Example 3: Preparation of in vivo tumor model by human glioma cell U87

[0177] Specific experimental steps:

[0178] Human glioma cell line U87 cells were cultured in DMEM medium containing 10% fetal bovine serum, and the medium was replaced every 2-3 days. When the cells were in the logarithmic growth phase, they were routinely centrifuged and collected for later use.

[0179] Sodium alginate and 15% gelatin solutions with mass volume fractions of 2% and 15% respectively were prepared and sterilized under high temperature and high pressure. First place the 8×10 6 / mL U87 cells were resuspended in 1mL cell culture medium, and then mixed with 2% sodium alginate and 15% gelatin solution at a volume ratio of 1:1:2 to obtain a final concentration of 7.5% gelatin, 0.5% sodium alginate and 2×10 6 / mL mixture of U87 cells.

[0180] A 3D multi-nozzle bioprinter was used to print a cube grid support with a side length of 20 mm and a thickness of 1.5 mm, with a filling rate ...

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Abstract

The invention relates to a 3D (three-dimensional) bioprinting in-vivo tumor model and a building method thereof, and belongs to the technical field of biology or the field of biological 3D printing. The tumor model comprises tumor cells and a hydrogel stent, wherein the hydrogel stent is a 3D stent obtained in a 3D bioprinting mode; and the hydrogel stent is transplanted into an animal body to form the in-vivo tumor model. The building method of the in-vivo tumor model comprises the steps of mixing, printing, hydrogel stent culture and transplantation. The invention aims at providing the high-tumorigenic-rate in-vivo tumor model capable of being fast and stably prepared in large scales, and the building method of the in-vivo tumor model. The tumor cell carrying hydrogel stent can be fast and stably prepared in large scales by using a 3D bioprinting technology; after in vitro culture for a short time, the tumor cell carrying hydrogel stent is transplanted into a nude mouse body to obtain a high tumorigenic rate; tumor models of different parts and different types can be built according to research requirements; and the tumor models are built in a personalized manner.

Description

technical field [0001] The invention belongs to the field of biotechnology or the field of three-dimensional biological printing, and in particular relates to a three-dimensional bioprinted in vivo tumor model and a construction method thereof. Background technique [0002] Athymic nude mouse animal models have been widely used in research work in various fields such as oncology, immunology, and toxicology, which can help us study the mechanism of disease occurrence and development, screening of anticancer drugs, and improve the diagnosis and treatment of diseases. is of great significance. At present, the most widely used animal model of nude mice is the xenograft tumor model. Since nude mice lack thymus and lack mature T lymphocytes, the tumor grows well after transplantation, so it is an ideal and commonly used animal experiment tool in tumor research. According to the different transplantation methods, it can be divided into tumor tissue block transplantation and tumor ...

Claims

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Application Information

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IPC IPC(8): C12N5/09A01K67/027
CPCC12N5/0693A01K67/0271C12N2533/74C12N2533/54C12N2533/80C12N2533/70A01K2207/12A01K2227/105A01K2267/0331
Inventor 王宣之龙小燕
Owner THE FIRST AFFILIATED HOSPITAL OF WANNAN MEDICAL COLLEGE YIJISHAN HOSPITAL OF WANNAN MEDICAL COLLEGE
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