Efficient cell culture microcarrier in field of bioengineering

Abstract

The utility model relates to the technical field of bioengineering, and discloses an efficient cell culture microcarrier in the field of bioengineering, which comprises a carrier body, the carrier body consists of a support layer, a culture layer and an adhesion layer, and the support layer, the culture layer and the adhesion layer are sequentially adhered together in pairs through an adhesive. The supporting layer is located on the lower portion of the culture layer, the adhesion layer is located on the upper portion of the culture layer, and the supporting layer, the culture layer and the adhesion layer are all cylinders with the thickness of 5-20 mm. The supporting layer is made of one or more compositions of dextran, synthetic polymer, glass, polyurethane foam and alginate gel, the cross section of the supporting layer is of a honeycomb hole structure, the supporting layer, the culture layer and the adhesion layer are arranged, connection is conducted through the adhesive, the structure is simple, cost is low, the structure is simple, and the culture medium is suitable for large-scale popularization and application. And under the matching action of the culture layer and the adhesion layer, the good survival of the cells can be ensured.

Description

TECHNICAL FIELD

[0001] The utility model relates to the field of bioengineering technology, specifically to a high -efficient cell culture microcarrier in the field of bioengineering. BACKGROUND

[0002] Microcarrier culture: microcarrier takes the micro particle as the carrier of cell adhesion, can provide considerable adhesion area, because the carrier volume is very small, the specific gravity is lighter, can make cell suspension in culture solution under slight stirring, finally can make cell reproduction into monolayer on the carrier surface one kind of cell culture technology. In practical operation, animal cell culture needs to grow by adhering, but the specific surface area of the container (such as culture bottle, glass slide or other container) used in traditional cell culture is small, so the number of cells produced by conventional adherent cell culture is limited. And microcarrier as a micron level particle has high specific surface area, adherent cells can adhere and grow on its surface, and can achieve the yield of several million cells per milliliter by stirring and suspending in the culture solution. In addition, the microcarrier culture technology can provide various forms of cell culture methods, such as suspension culture, perfusion culture and well plate culture, which makes animal cell culture have more selectivity and is more economical and feasible.

[0003] At present, the commonly used matrix of commercial microcarrier is dextran, cellulose, polyethylene or other polymers, etc. For some cells with low adhesion force and difficult to adhere in vitro, the commonly used microcarrier is prepared by covalently crosslinking animal gelatin protein-porcine collagen protein on the matrix, and amino acids, complex ion ligand, polypeptide or artificially synthesized polypeptide have similar properties to animal gelatin protein, which can provide a good adherent growth environment for cells and will not introduce animal-derived components in cell culture. Therefore, microcarriers with amino acids or polypeptides on the surface or how to prepare suitable ligands according to actual needs have become a hot spot in the field. In order to improve the adhesion speed and growth rate of animal cells on the surface of microcarrier and overcome the safety problem caused by animal-derived components, there is an urgent need for new microcarriers containing both charge and amino acid or polypeptide in the market to establish an efficient and safe system and scheme in the whole cell culture process. UTILITY MODEL CONTENT

[0004] (I) Technical problem solved

[0005] The utility model aims at providing a high -efficient cell culture microcarrier in the field of bioengineering to solve the problems in the above background technology.

[0006] (II) Technical scheme

[0007] In order to achieve the above object, the utility model provides the following technical scheme:

[0008] The utility model provides a high -efficient cell culture microcarrier in the field of bioengineering, including carrier body, the carrier body is by support layer, culture layer and attachment layer constitute, support layer, culture layer and attachment layer are pasted together through adhesive two two, support layer is located in the lower part of culture layer, attachment layer is located in the upper part of culture layer, support layer, culture layer and attachment layer, support layer, culture layer and attachment layer all are the cylinder of thickness 5~20mm.

[0009] Preferably, the material of the support layer is one or more combinations of dextran, synthetic polymer, glass, polyurethane foam and alginate gel, and the cross section of the support layer is provided with a honeycomb hole structure.

[0010] Preferably, the cross section of the culture layer is provided with a regular octagonal hole at the center, a pentagonal hole is arranged around the side of the regular octagonal hole, and a fan-shaped grid is arranged outside the pentagonal hole.

[0011] Preferably, the regular octagonal hole is provided with a group, and the pentagonal hole is provided with eight groups, wherein each pentagonal hole is located at one side of the regular octagonal hole on the inside, and the inside edge of each fan-shaped grid is the side of the pentagonal hole located outside.

[0012] Preferably, the outer edge of each fan-shaped grid is in a circular arc wave shape, at least one vertical rib is arranged on the inside edge of the fan-shaped grid, and the rib is recessed into the support layer and the attachment layer.

[0013] Preferably, the attachment layer comprises a coarse fiber structure and a fine fiber structure, the fine fiber structure is arranged on the surface of the coarse fiber structure, and the number of layers of the coarse fiber structure and the fine fiber structure is 2-3.

[0014] Preferably, the extending directions of the fiber filaments of each layer of the coarse fiber structure are arranged parallel to each other, the extending directions of the fiber filaments of each layer of the fine fiber structure are arranged parallel to each other, and the extending directions of the fiber filaments of the two adjacent layers of the coarse fiber structure form an included angle, wherein the included angle is 30°-120°.

[0015] Compared with the prior art, the utility model has the beneficial effects that:

[0016] The utility model discloses, through the setting of support layer, culture layer and attachment layer, adopt adhesive two two paste together, then simple structure, cost is lower, simultaneously through the setting of fan -shaped grid, equivalent to adopt the wrinkle, set up a plurality of through -hole mode, increase the surface area of this microorganism carrier, thereby for the microorganism implantation provided with better basis, more favorable to the survival of microorganism, finally through the fibre silk extension direction between the included angle of the fibre silk of adjacent two layers of coarse fibre structure constitutes such setting, makes adjacent two layers of coarse fibre structure can form the net -like structure, more convenient for the growth of cell, and the fine fibre can also be set up as nanometer fibre, and the structure aspect is similar with the collagen and elastic fibre in mammal body, and therefore cell can very easily adhere to nanometer fibre. BRIEF DESCRIPTION OF DRAWINGS

[0017] Figure 1 It is the front view integral structure schematic drawing of a kind of high-efficiency cell culture microcarrier in biological engineering field of the utility model;

[0018] Figure 2 It is the support layer cross section structure schematic drawing of a kind of high-efficiency cell culture microcarrier in biological engineering field of the utility model;

[0019] Figure 3 It is the culture layer cross section structure schematic drawing of a kind of high-efficiency cell culture microcarrier in biological engineering field of the utility model;

[0020] Figure 4 It is the attachment layer side section structure schematic drawing of a kind of high-efficiency cell culture microcarrier in biological engineering field of the utility model; Figure 3 In A area enlarged structure schematic diagram;

[0021] Figure 5 It is the attachment layer side section structure schematic drawing of a kind of high-efficiency cell culture microcarrier in biological engineering field of the utility model;

[0022] In the drawing: 1, carrier body;11, support layer;12, culture layer;121, regular octagonal hole;122, pentagonal hole;123, fan-shaped grid;124, convex rib;13, attachment layer;131, coarse fibre structure;132, fine fibre structure. DETAILED DESCRIPTION

[0023] The technical solutions in the embodiments of the utility model will be clearly and completely described below with reference to the drawings in the embodiments of the utility model. Obviously, the described embodiments are only part of the embodiments of the utility model, not all the embodiments. Based on the embodiments in the utility model, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of the utility model.

[0024] Please refer to Figures 1-5The utility model provides a kind of efficient cell culture microcarrier technical scheme in bioengineering field:

[0025] An efficient cell culture microcarrier in bioengineering field, including carrier body 1, carrier body 1 is constituted by support layer 11, culture layer 12 and attachment layer 13, support layer 11, culture layer 12 and attachment layer 13 are pasted together in turn by adhesive, support layer 11 is located in the lower part of culture layer 12, attachment layer 13 is located in the upper part of culture layer 12, support layer 11, culture layer 12 and attachment layer 13, support layer 11, culture layer 12 and attachment layer 13 are all cylinder with thickness of 5~20mm.

[0026] Further, the material of support layer 11 is one or more compositions of dextran, synthetic polymer, glass, polyurethane foam and alginate gel, and the cross section of support layer 11 is set as a honeycomb hole structure.

[0027] It should be noted that dextran is a commonly used natural polysaccharide, which has good biocompatibility and biodegradability, and is used as a support material for microcarriers. It can provide a good cell attachment and growth environment. Synthetic polymers such as polystyrene and PHEMA have good chemical stability and are easy to process. By adjusting the chemical structure, the cell attachment and growth can be optimized. Glass materials are used to make microcarriers due to their transparency and chemical stability, which can facilitate the observation of cell growth under a microscope.

[0028] Further, the cross section of culture layer 12 is set as a regular octagonal hole 121 at the center, and a pentagonal hole 122 is arranged around the regular octagonal hole 121, and a fan-shaped grid 123 is arranged outside the pentagonal hole 122.

[0029] The regular octagonal hole 121 is arranged in one group, and the pentagonal hole 122 is arranged in eight groups. Each pentagonal hole 122 is located at one side of the inner regular octagonal hole 121, and the inner side of each fan-shaped grid 123 is the outer side of the adjacent pentagonal hole 122.

[0030] The outer edge of each fan-shaped grid 123 is in a circular arc wave shape, and at least one vertical rib 124 is arranged on the inner side of the fan-shaped grid 123, and the rib 124 is recessed into the support layer 11 and the attachment layer 13.

[0031] It should be noted that by arranging the fan-shaped grid 123, it is equivalent to using wrinkles, arranging a plurality of through holes and other methods to increase the surface area of the microorganism carrier, thereby providing a good foundation for the adhesion of microorganisms, which is beneficial to the survival of microorganisms. By arranging the rib 124, the connection performance between the culture layer 12 and the support layer 11 and the attachment layer 13 is greatly enhanced.

[0032] Further, the attachment layer 13 comprises a coarse fiber structure 131 and a fine fiber structure 132, the fine fiber structure 132 is arranged on the surface of the coarse fiber structure 131, the number of layers of the coarse fiber structure and the fine fiber structure ranges from 2 to 3;

[0033] The extending direction of the fiber filaments of each layer of the coarse fiber structure 131 is arranged in parallel with each other, the extending direction of the fiber filaments of each layer of the fine fiber structure 132 is arranged in parallel with each other, and the extending direction of the fiber filaments of the two adjacent layers of the coarse fiber structure 131 forms an included angle, wherein the included angle ranges from 30° to 120°.

[0034] It should be noted that the two adjacent layers of the coarse fiber structure 131 are arranged in the form of the net structure by forming the included angle between the extending direction of the fiber filaments, which is more convenient for the growth of the cells, the fine fiber can also be arranged in the form of nanofiber, which is similar to the collagen and elastic fiber in the mammal body in terms of structure, so that the cells can be easily attached to the nanofiber, and the nanofiber can effectively promote the differentiation of the stem cells thereon, therefore, the addition of the fine fiber to the three-dimensional microcarrier can improve the cell inoculation efficiency, and can promote and regulate the growth and differentiation of the cells, especially the stem cells, grown thereon.

[0035] Finally, it should be noted that the above content is only used to illustrate the technical scheme of the present application, and is not a limitation on the protection scope of the present application, and the simple modification or equivalent replacement of the technical scheme of the present application by the ordinary skilled in the art does not deviate from the essence and scope of the technical scheme of the present application.

Claims

1. A high-performance cell culture microcarrier in the field of bioengineering, comprising a carrier body (1), characterized in that, The carrier body (1) is composed of a support layer (11), a culture layer (12) and an attachment layer (13), the support layer (11), the culture layer (12) and the attachment layer (13) are adhered together by adhesive in turn, the support layer (11) is located at the lower part of the culture layer (12), the attachment layer (13) is located at the upper part of the culture layer (12), the support layer (11), the culture layer (12) and the attachment layer (13) are all columnar bodies with a thickness of 5-20mm.

2. The high efficient microcarrier for cell culture in the field of bioengineering according to claim 1, characterized in that: The material of the support layer (11) is one or more combinations of dextran, synthetic polymer, glass, polyurethane foam and alginate gel, and the cross section of the support layer (11) is provided with a honeycomb hole structure.

3. The high efficient microcarrier for cell culture in the field of bioengineering according to claim 1, characterized in that: The cross section of the culture layer (12) is provided with a regular octagonal hole (121) at the center, a pentagonal hole (122) is arranged around the side of the regular octagonal hole (121), and a fan-shaped grid (123) is arranged outside the pentagonal hole (122).

4. The high-performance microcarrier for cell culture in the field of bioengineering according to claim 3, wherein: The regular octagonal hole (121) is provided with a group, and the pentagonal hole (122) is provided with eight groups, wherein each pentagonal hole (122) is located at one side of the regular octagonal hole (121) on the inside, and the inside edge of each fan-shaped grid (123) is the outside edge of the adjacent pentagonal hole (122).

5. The high-performance microcarrier for cell culture in the field of bioengineering according to claim 4, wherein: The outer edge of each fan-shaped grid (123) is in a circular arc wave shape, at least one vertical rib (124) is arranged on the inside edge of the fan-shaped grid (123), and the rib (124) is recessed into the support layer (11) and the attachment layer (13).

6. The high efficient microcarrier for cell culture in the field of bioengineering according to claim 1, wherein: The attachment layer (13) includes a coarse fiber structure (131) and a fine fiber structure (132), the fine fiber structure (132) is arranged on the surface of the coarse fiber structure (131), and the number of layers of the coarse fiber structure and the fine fiber structure is 2-3.

7. The high-performance cell culture microcarrier of claim 6, wherein the microcarrier is characterized by: The extension direction of the fiber silk of each layer of the coarse fiber structure (131) is arranged parallel to each other, the extension direction of the fiber silk of each layer of the fine fiber structure (132) is arranged parallel to each other, and the extension direction of the fiber silk of the adjacent two layers of the coarse fiber structure (131) forms an included angle, wherein the included angle is 30°-120°.

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