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Diphasic porous three-dimensional cell culture scaffold

A technology for three-dimensional cells and culturing scaffolds, which is applied in tissue culture, methods of supporting/fixing microorganisms, and biochemical instruments, etc., can solve the problems affecting the inoculation efficiency of cells in three-dimensional scaffolds, the difficulty of determining the pore size and the shape of the pores, and the difficulty of operation. , to achieve the effect of improving cell seeding efficiency, promoting and regulating growth and differentiation, and promoting cell adsorption

Inactive Publication Date: 2012-10-10
JIANGYIN RECONGENE BIOMEDICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the disadvantages of these fibrous polystyrene matrices are that the pore size and pore shape of the matrix are not easily determined and the fibrous matrix is ​​soft in nature, allowing the matrix to be deformed without deformation. Difficulty in performing further operations
However, during cell seeding, since the wells are 100% connected, a small amount of cells will leak from the three-dimensional cell culture scaffold to the cell culture plate, thereby affecting the seeding efficiency of cells on the three-dimensional scaffold

Method used

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  • Diphasic porous three-dimensional cell culture scaffold

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Example 1: Manufacturing method of biphasic porous three-dimensional cell culture scaffold

[0072] method one

[0073] (A) A biphasic porous three-dimensional cell culture scaffold was fabricated using polystyrene material, and a semi-finished porous three-dimensional scaffold composed of coarse fiber phase was prepared by rapid prototyping method.

[0074] (B) Electrospinning technology was used to spray fine fibers on the outer surface of the semi-finished porous three-dimensional scaffold.

[0075] (C) Continue to use the rapid prototyping method to prepare a porous three-dimensional scaffold on the surface with the thin fiber phase, so that the thin fiber phase is fixed inside the three-dimensional scaffold composed of the thick fiber phase.

[0076] (D) Using a mechanical cutting method. Cut the biphasic porous 3D cell culture scaffold to the desired size.

[0077] (E) Using a plasma surface treatment. The biphasic porous 3D cell culture scaffolds were plasma-...

Embodiment 2

[0085] Example 2: Using a biphasic porous three-dimensional cell culture scaffold to culture cells

[0086] The invention also provides a method for cultivating living cells using the biphasic porous three-dimensional cell culture scaffold in a tissue culture polystyrene plate. The biphasic porous three-dimensional cell culture scaffold used in this study has dimensions of 10 mm wide by 10 mm long by 0.3 mm thick, with square pores of 200 μm and fiber diameters of 400 μm. The fine fiber diameter is 1 μm.

[0087] Inoculate smooth muscle cells using the static inoculation method: Add 500 μl of smooth muscle cell suspension (1×105 cells / ml) from the upper surface of the scaffold with a pipette, allow the cells to attach for 2 hours at 37°C, and then pour into More cell culture media. After seeding the cells, the cell culture scaffolds were placed into multi-well plates containing cell culture medium and grown in an incubator at 37°C in an air atmosphere with 90% humidity and 5...

Embodiment 3

[0090] Example 3: Use with Polystyrene Tissue Culture Plates

[0091] The invention also provides a method for cultivating living cells in a polystyrene tissue culture plate by using the cell culture scaffold. The cell culture scaffold is a disc or cube shaped to fit the wells of a tissue culture plate. Cells are seeded in the cell culture scaffold using dynamic seeding or static seeding methods.

[0092] In one embodiment, using a static inoculation method, a certain volume of cell suspension is added from the upper surface of the cell culture support with a pipette, and the cells are allowed to attach to it for a certain period of time before being cultured. Fluid lavage. After inoculating the cells, put the cell culture scaffold into a well plate containing the cell culture solution, and culture it in an incubator at 37°C in an atmosphere with a relative humidity of 90% and a carbon dioxide content of 5-10%.

[0093] In another embodiment, a dynamic seeding method is use...

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Abstract

The invention relates to a diphasic porous three-dimensional cell culture scaffold which is formed by a coarse fiber phase and a fine fiber phase which are entirely different in diameter. The coarse fiber phase is larger than cultured cells in size while the fine fiber phase is smaller than the cultured cells in size, the coarse fiber phase comprises multilayer coarse fiber structures, each two adjacent coarse fiber structures are arranged according to a certain angle, and the fine fiber phase is individually combined on one side or multiple sides of the coarse fiber phase and is averagely or intensively distributed in hole structures of the three-dimensional cell culture scaffold formed by the coarse fiber phase. In the diphasic porous three-dimensional cell culture scaffold, the fine fiber phase is much smaller than the cells in diameter, so that the cells can attach to nanofibers quite easily, and differentiation of stem cells on the nanofibers can be effectively promoted. Therefore, by means of adding the fine fibers on the three-dimensional cell culture scaffold, cell vaccination efficiency can be improved, and the cells grown on the fine fibers, particularly growth and differentiation of the stem cells, can be promoted and regulated.

Description

technical field [0001] The invention relates to a two-phase porous three-dimensional cell culture support, which belongs to the field of cell and tissue culture. Background technique [0002] In the prior art, the main cell culture methods routinely used include the following two types: [0003] 1) Cultivate cells in a monolayer two-dimensional manner [0004] Cell culture is a very useful and widely used technique in the fields of drug discovery, cell biology, toxicology, bioengineering, and tissue engineering. Conventional cell culture is carried out in cell culture plates such as 2, 4, 6, 24, 96 well cell culture plates, which are made of non-degradable polymers including polystyrene. These cell culture plates are often treated with plasma to improve the hydrophilicity of the surface so that cultured cells can better adhere to the two-dimensional surface of the plate. In a typical cell culture experiment using polystyrene cell culture plates, cultured cells are grown i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00
CPCC12M25/14
Inventor 刘青
Owner JIANGYIN RECONGENE BIOMEDICAL TECH
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