Cell three-dimensional culture scaffold, preparation method and application thereof

A three-dimensional culture and cell technology, applied in the fields of biological tissue engineering and biomedicine, can solve the problems of poor cell infiltration effect, lack of precise control of freeze-drying and quenching process, lack of precise control of scaffold shape and parameter pore size, etc.

Active Publication Date: 2015-08-12
TIANJIN WEIKAI BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0019] However, the porous scaffolds currently prepared by freeze-drying methods have many shortcomings, for example: most of them are not prepared directly in porous cell culture dishes; most of them lack precise control over the freeze-drying and quenching process, so the morph...

Method used

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  • Cell three-dimensional culture scaffold, preparation method and application thereof
  • Cell three-dimensional culture scaffold, preparation method and application thereof
  • Cell three-dimensional culture scaffold, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0118] Example 1: Preparation of cell three-dimensional culture scaffold (porous scaffold)

[0119] 1. Solution preparation

[0120] 1. First use pure acetic acid and deionized water to prepare a 0.1M / L acetic acid solution (that is, a concentration of 0.6% v / v) in a sterilized container, and then filter it with a 0.22 μm filter membrane and store it in a closed sterile container (not Store in a 4°C refrigerator when in use);

[0121] 2. Use 0.1M / L acetic acid solution to dissolve chitosan (analytically pure, purified from shrimp and crab shells in nature, medium molecular weight range, such as Sigma417963) in a sterilized container, according to the concentration of 2% w / v chitosan The solution is prepared and the top of the container is closed with a cap. Place the solution to be dissolved in a 55°C water bath beaker (the level of the water bath must be higher than the level of the chitosan solution), and use a heatable magnetic stirrer to keep the temperature of...

Embodiment 2

[0141] Embodiment 2: cell culture experiment

[0142] 1. Purpose of the experiment

[0143] Evaluate the effect of chitosan-collagen on cell growth of the three-dimensional scaffold material (50 μl scaffold and 75 μl scaffold) prepared in Example 1, method: cell line HepG2 and primary cell adipose stem cells (cell line HepG2 is purchased from ATCC, adipose stem cell Commercially available or prepared according to conventional methods in the art) were inoculated on the scaffold material, and cultured in vitro for a week, and the number of cells in the scaffold was detected by CCK-8 at different time points.

[0144] 2. Experimental materials and equipment

[0145] As shown in Table 2 and Table 3 below.

[0146] Table 2: Experimental materials

[0147]

[0148]

[0149] Table 3: Experimental equipment

[0150] name

Manufacturer

model

Ultra low temperature centrifuge

Thermo

LEGEND Mach1.6R

Carbon dioxide incubator

STIK

...

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PUM

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Abstract

The invention belongs to the fields of biological tissue engineering and biological medicines and relates to a cell three-dimensional culture scaffold, a preparation method and an application thereof. In particular, one or more pores are distributed in the cell three-dimensional culture scaffold, wherein each pore can accommodate at least one cell; in addition, the pores have openings, through which at least one cell can enter the pores. The cell three-dimensional culture scaffold is compact in pore size, is excellent in biocompatibility and cell permeability, is good in strength and is free of collapse, separation and the like defects.

Description

technical field [0001] The invention belongs to the fields of biological tissue engineering and biomedicine, and relates to a three-dimensional cell culture support, its preparation method and application. Background technique [0002] A number of studies have shown that when cells grow in the human body, they are in a three-dimensional environment, and their shape and function are very different from those of single-layer two-dimensional cells cultured in vitro. It has been continuously documented that cells are significantly different in two-dimensional and three-dimensional culture methods. [0003] The usual two-dimensional cell culture is not the same as the growth mode of cells in human or animal body, and often cannot correctly reflect the structure or function under normal or pathological tissue conditions, so the results may be inaccurate when using this method for experiments such as drug screening . [0004] Compared with two-dimensional cell culture, three-dime...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12N5/0775C12N5/071A61L27/56A61L27/26A61L27/20A61L27/24C08J9/28
Inventor 刘仁辰魏搏超崔雪峰
Owner TIANJIN WEIKAI BIOENG
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