Microcarrier structure for 3D culture of cells

Abstract

The utility model discloses a microcarrier structure for 3D (three-dimensional) cell culture, belongs to the technical field of cell culture, and solves the problem that the center of a cell ball still lacks a growth microenvironment while the three-dimensional growth of the cell ball is realized by the existing cell culture device. The device comprises a sphere-like polyhedron with a hollow interior, the interior of the sphere-like polyhedron is filled with a culture medium, the plane of the sphere-like polyhedron comprises a solid plane, and a plurality of round holes penetrating through the solid plane are uniformly formed in the solid plane; the plane of the sphere-like polyhedron comprises at least one group of two cavity planes. The sphere-like polyhedron disclosed by the utility model is similar to a sphere and is beneficial to rapid and stable adhesion and growth of cells, and meanwhile, a culture medium can be filled in the hollow interior of the sphere-like polyhedron so as to simulate a central microenvironment when a cell cluster or a cell sphere is formed, so that the exchange of nutrition and oxygen of internal cells and the excretion of wastes are met; and the necrosis of cells in a cell cluster or a cell ball during 3D cell culture is avoided.

Description

Technical Field

[0001] The utility model relates to the technical field of cell culture, and particularly relates to a microcarrier structure for 3D cell culture. Background Art

[0002] Cell culture is a common technique for maintaining the growth of cells or cell populations under in vitro conditions. Traditionally, cells are amplified in a monolayer on two-dimensional (2D) culture plates / flasks / dishes. The advantage of 2D cell culture is that it provides an easy-to-adhere growth plane for cells. However, monolayer culture requires routine passage to maintain cell self-renewal and potential, which is limited for large-scale cell proliferation. Moreover, two-dimensional attachment changes the shape and geometric structure of cells, leading to changes in the overall cell, cytoskeleton, and nuclear morphology. Subsequently, the change in cell morphology affects gene and protein expression. In addition, the composition and components of the extracellular matrix (ECM) are extremely important for the function of cells. However, 2D culture often lacks the formation of ECM. Therefore, the 2D culture conditions lack the complexity required to simulate the cell microenvironment, dynamics, and specific three-dimensional (3D) space, that is, it is difficult to achieve the complex spatial interaction between cells and ECM components, nutrients, oxygen, and waste.

[0003] 3D cell culture is a technique that promotes the growth of cells in three-dimensional space in an artificially created environment. The 3D culture technique has the advantages of both retaining the material and structural basis of the in vivo cell microenvironment and demonstrating the intuitiveness and condition controllability of cell culture. Currently, 3D cell culture is mainly divided into two types: non-scaffold culture method and scaffold culture method.

[0004] Non-scaffold 3D culture uses techniques such as hanging drop microwell plates and magnetic nanoparticles to promote cell self-aggregation and the formation of spherical solid cell aggregates. The cells inside the cell aggregates formed by non-scaffold 3D culture are prone to necrosis due to lack of nutrients and oxygen, and it is difficult to simulate the real growth state of cells in vivo.

[0005] The scaffold culture method is a cell culture technique based on scaffold support, which can provide physical support, from simple mechanical structures to extracellular matrix analogs. Cells can aggregate, proliferate and migrate on the scaffold. 3D culture with scaffolds can be further divided into cell-embedded growth and supported growth. Embedded growth means that cells are embedded in the matrix, and the physicochemical properties of the matrix will affect the characteristics of the cells. This culture method has high requirements for the matrix and cannot be recycled, so the culture cost is greatly increased. During supported growth, cells attach to three-dimensional microcarriers to achieve three-dimensional growth. In suspension culture, they provide an adhesion surface for anchorage-dependent cells. The adhesion of cells on the microcarriers can be optimized to promote cell expansion or differentiation. Microcarriers provide a large surface area for cells to grow to high density, so that concentrated cells can be collected by enzymatic dissociation. However, although microcarriers expand the cell growth area and simulate the 3D cell growth space, they still have the following disadvantages: 1. Most microcarriers are solid beads or uneven porous beads. While realizing the three-dimensional growth of cell spheres, the growth microenvironment is still lacking in the center of the cell spheres; 2. Cells attach and grow on the microcarriers, but it is still difficult for the inner cells close to the microcarriers to achieve complex spatial interactions with ECM components, nutrients, oxygen, and waste. Summary of the Utility Model

[0006] In view of the above problems in the prior art, the present utility model provides a microcarrier structure for 3D cell culture, which solves the problem that the growth microenvironment is still lacking in the center of the cell sphere while realizing the three-dimensional growth of the cell sphere in the existing cell culture device.

[0007] In order to achieve the above object, the technical solution adopted by the present utility model is as follows:

[0008] Provide a microcarrier structure for 3D cell culture, which includes a spherical polyhedron with a hollow interior. The hollow interior of the spherical polyhedron is filled with a culture matrix. The planes of the spherical polyhedron include solid planes, and a number of round holes penetrating the solid planes are evenly arranged on the solid planes.

[0009] The spherical polyhedron of the present utility model is similar to a sphere, which is conducive to the rapid and stable adhesion and growth of cells. At the same time, the hollow interior of the spherical polyhedron can be filled with a culture matrix to mimic the central microenvironment during the formation of cell clusters or cell spheres, meeting the exchange of nutrients and oxygen and the excretion of waste of the inner cells, and avoiding the necrosis of the inner cells during 3D cell culture.

[0010] Further, the planes of the spherical polyhedron include at least one set of two hollow planes. The two hollow planes are arranged on the spherical polyhedron and are not adjacent to each other.

[0011] Further, the diameter of the round holes is 0.1 - 0.4 μm.

[0012] Furthermore, the spherical polyhedron is a regular polyhedron.

[0013] Furthermore, each plane of the spherical polyhedron is a regular polygon.

[0014] Furthermore, the diameter of the circumscribed circle of the spherical polyhedron is 50 - 300 μm, the wall thickness of the spherical polyhedron is 3 - 10 μm, and the number of planes on the spherical polyhedron is greater than or equal to 10.

[0015] Furthermore, the material of the spherical polyhedron is medical-grade polystyrene, gelatin, dextran, and collagen.

[0016] The present utility model discloses a microcarrier structure for 3D cell culture, and its beneficial effects are as follows:

[0017] 1. The spherical polyhedron of the present utility model is similar to a sphere, which is conducive to the rapid and stable adhesion and growth of cells. At the same time, the internal hollow of the spherical polyhedron can be filled with a culture matrix to mimic the central microenvironment during the formation of cell clusters or cell spheres, meeting the exchange of nutrients and oxygen and the excretion of waste for the internal cells, and avoiding the necrosis of the internal cells during 3D cell culture.

[0018] 2. The present utility model fills the internal hollow of the spherical polyhedron with a culture matrix to meet the spatial interaction between cells and ECM components during cell growth.

[0019] 3. The present utility model is evenly provided with a number of round holes on the solid plane, and the internal cells of the cell cluster or cell sphere can carry out complex spatial interactions of ECM components, nutrients, oxygen, and waste with the culture matrix filled in the internal hollow of the spherical polyhedron through the round holes, which is more conducive to the survival, growth, and metabolism of the internal cells, and also due to maintaining the normal formation and survival of the cell cluster or cell sphere.

[0020] 4. The present utility model symmetrically provides at least one set of two hole planes on the spherical polyhedron, and the two hole planes are not adjacent. The hole planes are conducive to the entry and update of the culture matrix, so that the culture matrix in the internal hollow of the spherical polyhedron remains sufficient and the cell waste is discharged in time. Description of the Drawings

[0021] Figure 1 It is a schematic structural diagram of a microcarrier structure for 3D cell culture of the present utility model.

[0022] Figure 2 It is a schematic structural diagram of the solid plane of the present utility model.

[0023] Among them, 1. Spherical polyhedron; 2. Solid plane; 3. Round hole; 4. Hole plane. Detailed Embodiment

[0024] The specific embodiments of the present utility model will be described to facilitate those skilled in the art of this technology to understand the present utility model. However, it should be clear that the present utility model is not limited to the scope of the specific embodiments. For those of ordinary skill in the art of this technology, as long as various changes are within the spirit and scope of the present utility model defined and determined by the appended claims, these changes are obvious, and all utility models created using the concept of the present utility model are within the scope of protection.

[0025] Reference Figure 1 - Figure 2 , which is a schematic structural diagram of a microcarrier structure for 3D cell culture in this embodiment. Its purpose is to solve the problem that in the existing cell culture device, while realizing the three-dimensional growth of cell spheres, the central microenvironment for the growth of cells in the center of the cell spheres is still lacking. The following will describe the specific structure in this embodiment in detail.

[0026] A microcarrier structure for 3D cell culture, which includes a spherical polyhedron 1 with a hollow interior.

[0027] Among them, a culture medium matrix is filled in the hollow interior of the spherical polyhedron 1. The culture medium matrix can be an extracellular matrix, nutrients, culture solution, or / and matrix gel to mimic the central microenvironment when cell clusters or cell spheres are formed, meet the exchange of nutrients and oxygen and the excretion of waste of the internal cells, and avoid the necrosis of the internal cells during 3D cell culture.

[0028] Specifically, the plane of the spherical polyhedron 1 includes a solid plane 2, and a number of round holes 3 penetrating the solid plane 2 are evenly opened on the solid plane 2. The diameter of the round holes 3 is 0.1 - 0.4 μm.

[0029] In this embodiment, the spherical polyhedron 1 of the present utility model is similar to a sphere. The solid plane 2 on the spherical polyhedron 1 is conducive to the rapid and stable adhesion and growth of cells. At the same time, the internal cells of the cell cluster or cell sphere can carry out complex spatial interactions of ECM components, nutrients, oxygen, and waste with the culture medium matrix filled in the hollow interior of the spherical polyhedron 1 through the round holes 3, which is more conducive to the survival, growth, and metabolism of the internal cells, and also due to maintaining the normal formation and survival of the cell cluster or cell sphere.

[0030] However, the number and diameter of the round holes 3 can be adjusted according to the specific area size of the solid plane 2 on the spherical polyhedron 1. In this embodiment, the diameter of the round holes 3 is 0.2 μm, and the number of the round holes 3 is 100.

[0031] Specifically, the material of the spherical polyhedron 1 can be composed of various materials, including medical-grade polystyrene, gelatin, dextran, or / and collagen, and has diverse porosity and morphology after synthesis.

[0032] Specifically, the plane of the spherical polyhedron 1 includes at least one set of two hollow planes 4. The two hollow planes 4 are disposed on the spherical polyhedron 1, and the two hollow planes 4 are not adjacent.

[0033] In this embodiment, at least one set of hollow planes 4 is provided on the spherical polyhedron 1. The number of a set of hollow planes 4 is two. The two hollow planes 4 are disposed on the spherical polyhedron 1, and the two hollow planes 4 are not adjacent. The number of sets of the hollow planes 4 on the spherical polyhedron 1 can be set according to actual needs. In this embodiment, one set of hollow planes 4 is provided on the spherical polyhedron 1. The two hollow planes 4 of one set are symmetrically disposed on the spherical polyhedron 1. Thus, the culture medium inside the spherical polyhedron 1 can enter and exit through the hollow planes 4 for renewal, so that the culture medium in the hollow part inside the spherical polyhedron 1 is sufficient and the cell waste can be discharged in time.

[0034] The spherical polyhedron 1 can be a regular polyhedron or an irregular polyhedron. Each plane of the spherical polyhedron 1 can be a regular polygon or an irregular polygon. That is, the spherical polyhedron 1 can be a regular dodecahedron or an icosahedron (Platonic solid), or can also be a polyhedron with non-uniform plane shapes, but the formed polyhedron must be similar to a sphere.

[0035] Specifically, the spherical polyhedron 1 is a regular polyhedron. Each plane of the spherical polyhedron 1 is a regular polygon. The diameter of the circumscribed circle of the spherical polyhedron 1 is 50 - 300 μm. The wall thickness of the spherical polyhedron 1 is 3 - 10 μm. The number of planes on the spherical polyhedron 1 is greater than or equal to 10.

[0036] In this embodiment, the spherical polyhedron 1 is a regular dodecahedron. The regular dodecahedron has 12 planes. Each plane is a regular pentagon. The side length of the regular pentagon is 100 μm. The area of each regular pentagon is about 0.017 mm 2 , and the surface area is about 0.2 mm 2 . The radius of the circumscribed sphere of the spherical micro-polyhedron 1 (regular dodecahedron) is about 140 μm. The thickness of each face is uniform, all being 5 μm.

[0037] The spherical micro-polyhedron 1 (regular dodecahedron) is hollow inside. The volume of the hollow part is about 5.6×10 6 μm 3 .

[0038] Each solid plane 2 of the spherical micro-polyhedron 1 (regular dodecahedron) is provided with the same number and size of round holes 3. The diameter of the round holes 3 is 0.2 μm. The number of the round holes 3 is 100. The opening positions are arranged neatly and uniformly.

[0039] The spherical micro-polyhedron (regular dodecahedron) has a set of two opposite hollow planes 4. The hollow planes 4 are not adjacent. The hollow planes 4 and the solid planes 2 have the same area, about 0.017 mm2 。

[0040] Although the specific embodiments of the utility model have been described in detail with reference to the accompanying drawings, it should not be construed as a limitation on the scope of protection of this patent. Within the scope described in the claims, various modifications and variations that can be made by those skilled in the art without creative efforts still fall within the scope of protection of this patent.

Claims

1. A microcarrier structure for 3D cell culture, characterized in that: It includes a spherical polyhedron (1) with a hollow interior; the hollow interior of the spherical polyhedron (1) is filled with a culture medium matrix; The plane of the spherical polyhedron (1) includes a solid plane (2), and a number of circular holes (3) penetrating the solid plane (2) are evenly opened on the solid plane (2).

2. The microcarrier structure for 3D cell culture according to claim 1, characterized in that: The plane of the spherical polyhedron (1) includes at least one set of two hollow planes (4), and the two hollow planes (4) are arranged on the spherical polyhedron (1), and the two hollow planes (4) are not adjacent to each other.

3. The microcarrier structure for 3D cell culture according to claim 1, characterized in that: The diameter of the circular hole (3) is 0.1 - 0.4 μm.

4. The microcarrier structure for 3D cell culture according to claim 1, characterized in that: The spherical polyhedron (1) is a regular polyhedron.

5. The microcarrier structure for 3D cell culture according to claim 1, characterized in that: Each plane of the spherical polyhedron (1) is a regular polygon.

6. The microcarrier structure for 3D cell culture according to claim 1, characterized in that: The diameter of the circumscribed circle of the spherical polyhedron (1) is 50 - 300 μm, and the wall thickness of the spherical polyhedron (1) is 3 - 10 μm; the number of planes on the spherical polyhedron (1) is greater than or equal to 10.

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