Microculture tank structure for cell culture and cell culture vessel

Abstract

Microculture tank structure for cell culture and cell culture vessel. The microculture tank structure includes a microculture tank defining a micro culture space that opens upwards, and a microstructure unit disposed in the microculture tank and located in the micro culture space. The microstructure unit includes a plurality of microstructures extending upwardly protruding from the inner tank of the micro culture space. The microstructures cooperatively define a concave surface that tapers downward and that can be used to bear cells. Via the structural designs of the microculture tank and the microstructure unit of each of the microculture tank structure, can allow the cells in culture to slide naturally downward to the center of the concave surface. The microculture tank structure is adapted to expand the cells along the center of the concave surface and gradually aggregate into a spherical bulk form.

Description

Technical Field

[0001] This invention relates to a culture vessel, and more particularly to a microtrough structure and cell culture vessel for cell culture. Prior Technology

[0002] Traditional cell culture methods involve culturing cells in culture flasks or dishes, allowing them to grow and distribute on a flat surface. Therefore, this method can only produce a monolayer of cells. The cell morphology and function of cells cultured using this method differ significantly from those of living organisms.

[0003] In recent years, cell culture has gradually evolved from two-dimensional planar culture to three-dimensional spatial culture, which allows cultured cells to aggregate and grow, thus simulating organ tissues and enabling various biological simulation experiments, such as drug trials and cancer cell research, which contributes to the development of regenerative medicine and medical research.

[0004] Currently known three-dimensional cell culture techniques can be broadly categorized into scaffold-based methods and scaffold-free approaches.

[0005] The scaffold method uses a three-dimensional biopolymer with a mesh-like structure as a scaffold to simulate the extracellular matrix environment, allowing cells to attach and grow. However, this three-dimensional scaffold structure restricts cell imaging and hinders observation.

[0006] The scaffold-free method involves cell culture without physical support, employing methods such as magnetic levitation and non-adhesive culture with surface coatings. Magnetic levitation uses magnetically labeled nanomolecules to tag cells, then applies magnetic force to induce cell aggregation and growth in the culture medium; therefore, this method requires prior magnetic labeling of the cells. Non-adhesive culture with surface coatings involves applying a coating to the surface of a culture tank structure with a circular, conical, or V-shaped bottom. This reduces the contact area between the cells and the culture structure surface, causing the cells to remain suspended and easily aggregate into spherical shapes. However, this method requires special surface coating treatment of the culture structure surface; the coating used must be non-toxic and non-degradable.

[0007] As can be seen from the above, the current three-dimensional cell culture technology still has inconveniences in its use and there is still room for improvement. Summary of the Invention

[0008] Therefore, the object of the present invention is to provide a microgroove structure for cell culture that can improve upon at least one of the disadvantages of prior art.

[0009] Therefore, the microtrough structure of the present invention is suitable for cell culture. The microtrough structure includes a microtrough body defining an upward-facing micro-culture space, and a microstructural unit disposed in the microtrough body and located in the micro-culture space.

[0010] The microstructure unit comprises multiple microstructures extending longitudinally and protruding from the edge of the groove within the micro-culture space. These microstructures cooperate to define a support concave surface with a gradually narrowing diameter from top to bottom, which can be used to support the cells.

[0011] Another object of the present invention is to provide a cell culture container that can improve upon at least one of the disadvantages of the prior art.

[0012] Therefore, the cell culture container of the present invention is suitable for cell culture. The cell culture container includes a substrate and a plurality of cell culture tank units arranged at intervals on the substrate. Each cell culture tank unit includes a main tank extending downward from the top surface of the substrate with its opening facing upward, and at least one microtrough structure disposed in the main tank for cell culture.

[0013] Each microgroove structure has a microgroove body recessed within the concave bottom edge of the corresponding main groove body, and a microstructural unit disposed within the microgroove body. The microgroove body defines an upward-opening micro-culture space. The microstructural unit is located within the micro-culture space and includes multiple microstructures extending vertically and protruding vertically from the groove edge within the micro-culture space. These microstructures cooperate to define a support concave surface with a gradually narrowing diameter from top to bottom, which can be used to support the cells.

[0014] The advantage of this invention is that, through the structural design of the micro-groove body and the micro-structure unit of each micro-groove structure, the cells in culture can naturally slide down to the center of the concave bottom of the supporting concave surface, so that the cells can proliferate and gradually aggregate into a spherical shape at the center of the concave bottom of the supporting concave surface. Simple Explanation of the Diagram

[0015] Other features and effects of the present invention will be clearly presented in the embodiments with reference to the drawings, wherein: Figure 1 is a perspective view illustrating the structure of a first embodiment of the cell culture container of the present invention; Figure 2 is an incomplete cross-sectional view, illustrating the structure of a cell culture tank unit of the first embodiment; Figure 3 is an incomplete perspective view, illustrating one of the microgroove structures of the first embodiment, and showing that multiple microstructures of the microgroove structure are hexagonal prisms; Figure 4 is a view similar to Figure 3, illustrating one of the microgroove structures of the first embodiment, and showing that the multiple microstructures of the microgroove structure are hexagonal tubes; Figure 5 is a view similar to Figure 2, illustrating another embodiment of the cell culture tank unit of the first embodiment; Figure 6 is an incomplete cross-sectional view, illustrating the structure of the cell culture tank unit of the first embodiment, wherein the microstructure is a hexagonal prism; Figure 7 is a three-dimensional diagram illustrating the arrangement of multiple microstructures in the shape of hexagonal prisms; Figure 8 is a view similar to Figure 3, illustrating that the multiple microstructures of the microgroove structure in the first embodiment are hexagonal tubes; Figure 9 is a plan view illustrating a honeycomb pattern formed by multiple adjacent hexagonal tubular microstructures. Figure 10 is a view similar to Figure 6, illustrating another embodiment of the microstructure described in the first embodiment; Figure 11 is a flowchart illustrating how, when a microgroove structure of the first embodiment is used to culture cells, the cells tend to clump together at the bottom of a support concave surface. Figure 12 is an incomplete perspective view, illustrating another embodiment of the microgroove structure of the first embodiment; Figure 13 is an incomplete cross-sectional view illustrating the structure of the embodiment shown in Figure 12; Figure 14 is an incomplete cross-sectional view, illustrating another embodiment of the cell culture tank unit of the first embodiment; Figure 15 is an incomplete perspective view, illustrating the structure of a cell culture tank unit according to a second embodiment of the cell culture container of the present invention; Figure 16 is an incomplete cross-sectional view, illustrating the structure of the cell culture tank unit shown in Figure 15; Figure 17 is an incomplete cross-sectional view, schematically illustrating the structure of a cell culture tank unit according to a third embodiment of the cell culture container of the present invention; and Figure 18 is an incomplete top view, illustrating the structure of the cell culture tank unit shown in Figure 17. Implementation

[0016] Before the present invention is described in detail, it should be noted that similar elements are represented by the same numbers in the following description.

[0017] Referring to Figures 1, 2, 3, and 4, a first embodiment of the cell culture container 200 of the present invention is shown, which is suitable for holding culture medium and can be used for cell culture. The cell culture container 200 includes a substrate 3 and a plurality of cell culture tank units 4 spaced apart and recessed on the top surface of the substrate 3.

[0018] Each cell culture tank unit 4 includes a main tank body 41 that extends downward from the top surface of the substrate 3 and has an inner concave bottom edge 410, a plurality of micro-groove structures 42 that are spaced apart and recessed in the inner concave bottom edge 410, and a plurality of surface microstructures 45 that protrude from the inner concave bottom edge 410.

[0019] It must be noted that the micro-groove structures 42 and surface microstructures 45 of the cell culture tank unit 4 are all in the micrometer to nanometer scale. Therefore, the illustrated style and size are only for illustration and should not be limited to the illustrated style in practice.

[0020] The main culture tank 41 defines a main culture space 411 that extends vertically and connects the microgroove structures 42, with its opening facing upwards. The concave bottom edge 410 defines a spacer structure 412 that separates the microgroove structures 42. The top surface of the spacer structure 412 can be flat as shown in FIG2, or convex upwards as shown in FIG5. In this first embodiment, the diameter of each main culture space 411 is 6 mm, but in practice, the diameter of the main culture space 411 is not limited to this.

[0021] The surface microstructures 45 are perpendicular to the top surface of the spacer structure 412 and are upright as shown in Figure 2, or extend upward and downward as shown in Figure 5. In addition, in accordance with the shape of the spacer structure 412, the top edges of the surface microstructures 45 can be distributed at the same height as shown in Figure 2, or distributed in an upward arc-shaped distribution as shown in Figure 5.

[0022] Referring to Figures 3 and 7, in this first embodiment, the surface microstructures 45 are polygonal prisms erected at intervals within the spacer structure 412. In this first embodiment, the surface microstructures 45 are designed as hexagonal prisms, and they are arranged in a honeycomb pattern. The maximum diameter (diagonal diameter) of each surface microstructure 45 is DO, and its vertical extension length is DL. The aspect ratio (DL / DO) is AR. The center-to-center distance (P) between two adjacent surface microstructures 45 is 1.1DO to 3DO, and AR is between 0.1 and 15. In this first embodiment, DO is between 200 nm and 3.5 μm, and DL is between 20 nm and 10 μm. In practice, in the above-described prism embodiment, each surface microstructure 45 can also be modified to, for example, but not limited to, triangular prisms, quadrilateral prisms, pentagonal prisms, or cylinders.

[0023] Referring to Figures 4 and 9, in another embodiment of the present invention, the surface microstructures 45 can also be replaced with hexagonal tubes, and the surface microstructures 45 are connected and arranged in a honeycomb pattern. The maximum inner diameter (diagonal inner diameter) of each microstructure 441 is Di, and the vertical extension length is DL. The aspect ratio (DL / Di) is AR. The center-to-center distance (P) between two adjacent microstructures 441 is 1.1Di to 3Di. [], AR = 0.1~15. In this first embodiment, Di is between 200 nm and 3.5 μm, and DL is between 20 nm and 10 μm. In practice, in the above-described tubular structure embodiment, each of the surface microstructures 45 can also be replaced with, for example, but not limited to, triangular tubes, quadrilateral tubes, pentagonal tubes, or circular tubes.

[0024] Since the microgroove structures 42 are identical, for ease of explanation, only one of them will be used as an example for structural description below.

[0025] Referring to Figures 2, 3, and 6, the microgroove structure 42 has a microgroove body 43 recessed in the corresponding concave bottom edge 410, and a microstructure unit 44 disposed on the inner edge of the microgroove body 43.

[0026] The micro-groove 43 has an annular opening wall 431 extending downward from the concave bottom edge 410, and a spherical bottom wall 432 connected to the periphery of the bottom edge of the opening wall 431 and having an upward-facing opening. The opening wall 431 and the bottom wall 432 cooperate to define a micro-culture space 430 with an upward-facing opening that connects to the main culture space 411.

[0027] In this first embodiment, the opening wall portion 431 is a vertically extending annular shape (as shown in FIG2). However, in other embodiments of the present invention, the opening wall portion 431 may also be a circular conical annular shape with a diameter gradually narrowing from top to bottom (as shown in FIG5). When the opening wall portion 431 is a truncated conical annular shape, the vertical inclination angle of the inner surface of the opening wall portion 431 may be between 5 degrees and 89 degrees.

[0028] Referring to Figures 3 and 6, the microstructure unit 44 includes a plurality of microstructures 441 protruding from the inner edges of the opening wall 431 and the bottom wall 432 of the groove. The microstructures 441 protruding from the opening wall 431 may extend laterally as shown in Figure 2 or Figure 5. The microstructures 441 protruding from the bottom wall 432 extend longitudinally vertically.

[0029] The microstructures 441 protruding from the bottom wall 432 of the groove will cooperate to define an upward-facing support concave surface 442, which can be used to support the cells 800. In this first embodiment, the support concave surface 442 is spherical with a curvature between 50 μm and 5 cm, and the opening diameter of the support concave surface 442 is between 10 µm and 1000 µm.

[0030] In practice, these microstructures 441 can have the following two structural forms. Structural type 1: Polygonal prism.

[0031] Referring to Figures 3, 6, and 7, each of the microstructures 441 is a polygonal prism with multiple sides. These microstructures 441 are spaced apart from each other, and adjacent microstructures 441 face each other with one of their sides spaced apart. The microstructures 441 protruding from the bottom wall portion 432 define the bearing concave surface 442 by their top surfaces engaging.

[0032] In this embodiment, each microstructure 441 is exemplified by a hexagonal prism. The maximum diameter (diagonal diameter) of each microstructure 441 is DO, and its vertical extension length is DL. The aspect ratio (DL / DO) is AR. The center-to-center distance (P) between two adjacent microstructures 441 is 1.1DO~3DO, and AR is between 0.1~15. In this first embodiment, DO is between 200 nm and 3.5 μm, and DL is between 20 nm and 10 μm. However, in other embodiments of the present invention, each microstructure 441 may also be a triangular prism, a quadrilateral prism, an octagonal prism, or a cylinder, etc. Structural type 2: Polygonal tube.

[0033] Referring to Figures 4, 8, and 9, each of the microstructures 441 is a polygonal tube. These microstructures 441 are adjacent and connected, and the microstructures 441 protruding from the bottom wall portion 432 define the bearing concave surface 442 by their top edges fitting together.

[0034] In this first embodiment, each microstructure 441 is a hexagonal tube, and these microstructures 441 are adjacent and connected in a honeycomb pattern. The maximum inner diameter (diagonal inner diameter) of each microstructure 441 is Di, and the downward extension length is DL. The aspect ratio (DL / Di) is AR. The center-to-center distance (P) between two adjacent microstructures 441 is 1.1Di to 3Di. [], AR = 0.1~15. In this first embodiment, Di is between 200 nm and 3.5 μm, and DL is between 20 nm and 10 μm. In other embodiments of the present invention, each microstructure 441 can also be replaced with a triangular tube, a quadrilateral tube, a pentagonal tube, or a circular tube.

[0035] In the two structural forms described above, the microstructures 441 can extend vertically upward from the inner edge of the bottom wall portion 432 of the groove, as shown in Figure 6, or they can extend vertically upward from the inner edge of the bottom wall portion 432 of the groove, as shown in Figure 9, with some parts being upright and others being inclined vertically.

[0036] Referring to Figures 2, 6, and 11, through the structural design of the two types of microgroove structures 42 described above, when a culture medium containing cells 800 is injected into one of the main grooves 41, the cells 800 will disperse and settle into the micro-culture spaces 430 defined by the microgroove bodies 43. When cells 800 fall into one of the micro-culture spaces 430, because the bearing concave surface 442 formed by the micro-nano-sized microstructures 441 is a porous surface with an arc-shaped concave shape, the cells 800 are not easily adhered to the bearing concave surface 442. The cells 800 will slide and concentrate towards the central area of ​​the concave bottom of the bearing concave surface 442 due to gravity. With this design, the cells 800 can proliferate in the central area of ​​the concave bottom of the bearing concave surface 442 and aggregate into a spherical shape due to gravity.

[0037] Referring to Figures 3, 12, and 13, in this first embodiment, the opening wall 431 of each microgroove 43 is annular, and the bottom wall 432 is spherical (as shown in Figure 3). However, in other embodiments of the present invention, the opening wall 431 of each microgroove 43 can be designed as a polygonal annular shape (as shown in Figure 12), such as, but not limited to, a triangular annular shape, a quadrilateral annular shape, and a hexagonal annular shape. The bottom wall 432 of each microgroove 43 is designed to correspond to the shape of the opening wall 431, and is changed to a polygonal cone shape with the opening gradually narrowing downwards and facing upwards, such as a triangular cone (as shown in the figure), a quadrilateral cone (as shown in the figure), and a hexagonal cone (as shown in the figure). The bearing concave surface 442 formed by the microstructures 441 provided on the bottom wall portion 432 of the pyramidal groove will also be in the shape of a polygonal pyramid, and the upward and downward inclined extension angle of the bearing concave surface 442 can be between 10 degrees and 75 degrees.

[0038] Furthermore, in this first embodiment, each main groove 41 is provided with a plurality of microgroove structures 42, for example, but not limited to 55. However, in another embodiment of the present invention, each main groove 41 may be provided with only a single microgroove structure 42.

[0039] Referring to Figure 14, in an embodiment where only one micro-groove structure 42 is provided in each main tank 41, the micro-groove body 43 of the micro-groove structure 42 may not have an opening wall, and the top edge of the bottom wall 432 is directly connected to the bottom edge of the inner periphery of the main tank 41, and the micro-culture space 430 is directly defined by the bottom wall 432, so that the opening diameter of the micro-culture space 430 is equal to the opening diameter of the main culture space 411.

[0040] With this design, the cells placed in each of the main tanks 41 will sink directly into the micro-culture space 430 of the micro-tank 43, and guided by the microstructures 441, they will gather and grow into clusters in the central area of ​​the concave bottom of the bearing concave surface 442.

[0041] Furthermore, through the structural design of the surface microstructures 45 in the spacer structure 412, when the surface microstructures 45 are distributed in a planar shape as shown in Figure 2, the cells settled on the surface microstructures 45 are less likely to adhere and stick, making it easier to allow the cells to fall into one of the adjacent microgrooves 42 by vibrating the entire cell culture container 200. When the surface microstructures 45 are distributed in an arc-convex shape as shown in Figure 5, the cells settled on the surface microstructures 45 are easily guided downwards into one of the adjacent microgrooves 42. However, in other embodiments of the present invention, it is not necessary to provide the surface microstructures 45 in the spacer structure 412.

[0042] Referring to Figures 15 and 16, the difference between a second embodiment of the cell culture container 200 of the present invention and the first embodiment lies in the structural design of the micro-groove body 43 of each micro-groove structure 42. For ease of explanation, the following description will only focus on the differences between the various embodiments, and the structure will be described using only a single micro-groove body 43 as an example.

[0043] In this second embodiment, the bottom wall portion 432 of the microgroove 43 has a polygonal annular segment 433 whose diameter gradually narrows from top to bottom, and a spherical bottom portion 434 connected to the bottom edge of the narrow annular segment 433 and having an upward-facing opening. In practice, the opening wall portion 431 is designed to correspond to the shape of the narrow annular segment 433 of the bottom wall portion 432, and thus has a polygonal annular shape corresponding to the narrow annular segment 433.

[0044] The microstructures 441 arranged on the inner edges of the narrow annular section 433 and the bottom section 434 will cooperate to define the bearing concave surface 442. Therefore, the bearing concave surface 442 has a spherical gathering area 443 located in the bottom section 434 with its opening facing upward, and a polygonal conical guiding area 444 located in the narrow annular section 433 and extending obliquely upward and outward from the periphery of the gathering area 443. In practice, the microstructures 441 can be various structural forms described in the first embodiment, such as spaced-apart columns or adjacent tubes.

[0045] During implementation, the vertical tilt angle of each side of the guide area 444 can be between 5 degrees and 135 degrees, and the vertical extension length of each side can be between 0.05 mm and 10 mm. The aggregation area 443 is hemispherical, and its diameter can be between 0.02 mm and 2.5 mm. The depth from the opening of the microgroove 43 to the lowest point of the aggregation area 443 can be between 0.1 mm and 1.5 mm.

[0046] Through the structural design of the bottom wall portion 432 of the micro-channel 43, and the structural design of the microstructures 441 arranged in the narrow ring segment 433 and the bottom segment 434 of the bottom wall portion 432, the cells falling into the micro-culture space 430 can also be guided to the aggregation area 443 of the bearing concave surface 442 by the microstructures 441, and grow into a ball shape in the aggregation area 443 due to gravity.

[0047] Referring to Figures 17 and 18, a third embodiment of the cell culture container of the present invention differs from the first embodiment in that the structural design of the cell culture tank unit 4 is as follows. The following description uses only a single cell culture tank unit 4 as an example.

[0048] In this third embodiment, the main tank 41 of the cell culture tank unit 4 contains only a single microtrough structure 42. The microtrough body 43 of the microtrough structure 42 includes only the bottom wall portion 432. The bottom wall portion 432 is connected to the bottom edge of the inner periphery of the main tank 41, and the opening diameter of the micro-culture space 430 defined by the bottom wall portion 432 is equal to the bottom diameter of the main culture space 411.

[0049] The bearing concave surface 442 of the microstructural unit 44 has a gathering area 443 located in the central region and an annular guiding area 444 that is higher than the gathering area 443 and extends outward from the periphery of the gathering area 443 in an upward arc. The microstructures 441 of the microstructural unit 44 are divided into a plurality of first microstructures 441' that cooperate to define the gathering area 443 and a plurality of second microstructures 441" that cooperate to define the guiding area 444. The top of the first microstructures 441' is lower than the top of the second microstructures 441".

[0050] In this third embodiment, the diameter of each main culture space 411 is 6 mm, and the depth from the opening of the main culture space 411 to the lowest point of the aggregation region 443 is 10.5 mm. The first microstructures 441' are hexagonal tubes that are adjacent to each other and connected in a honeycomb pattern, and the second microstructures 441" are hexagonal prisms that are spaced apart from each other and arranged in a honeycomb pattern. The area of ​​the aggregation region 443 can range from 0.01 to 20 mm2.

[0051] In practice, in other embodiments of the present invention, the first microstructures 441' and the second microstructures 441" may also be various structural types as described in the first embodiment, and the structural types of the first microstructures 441' of the aggregation region 443 and the second microstructures 441" of the guiding region 444 can be interchanged.

[0052] Through the structural design of the micro-groove structure 42 in each cell culture tank unit 4, when the main tank 41 is injected with culture medium containing cells 800, the cells 800 will directly sink into the micro-groove body 43 of the micro-groove structure 42. When cells 800 fall into the micro-culture space 430 of the micro-groove body 43, because the guide area 444 formed by the cooperation of the second micro-structures 441" is a porous surface that slopes downward from the periphery to the center, the cells 800 are not easily attached to the guide area 444, and the cells 800 will slide down to the aggregation area 443 due to gravity. And because the aggregation area 443 is also a porous surface that slopes downward from the periphery to the center, the cells 800 will also move and concentrate towards the center of the concave bottom of the aggregation area 443. With this design, the cells 800 can gradually proliferate at the center of the concave bottom of the aggregation area 443 and aggregate into a spherical shape due to gravity.

[0053] In summary, through the structural design of the micro-groove body 43 and the microstructure unit 44 of each microgroove structure 42, when the cells 800 in the culture medium fall into the micro-groove body 43, the supporting concave surface 442 formed by the micro-nano-sized columnar or tubular microstructures 441 is porous. Therefore, the adhesion of the cells 800 to the supporting concave surface 442 can be greatly reduced. In other words, the supporting concave surface 442 has very low adhesion to the cells 800. The cells 800 will naturally slide down the supporting concave surface 442 to the center of the concave bottom due to gravity, causing the cells 800 to proliferate and gradually aggregate into a spherical shape at the center of the concave bottom of the supporting concave surface 442. Therefore, the cell culture container 200 of the present invention is indeed a highly innovative creation and can indeed achieve the purpose of the present invention.

[0054] However, the above description is merely an embodiment of the present invention and should not be construed as limiting the scope of the present invention. Any simple equivalent changes and modifications made in accordance with the scope of the patent application and the contents of the patent specification of the present invention shall still fall within the scope of the patent of the present invention.

[0055] 200: Cell culture container 3:Substrate 4: Cell Culture Tank Unit 41: Main tank 410: Concave bottom edge 411: Main Incubation Space 412: Spacing structure 42: Microgroove structure 430: Micro-culture space 43: Micro groove body 431: Opening wall 432: Bottom wall of the tank 433: Narrowing loop segment 434: Bottom section of the trench 44: Microstructural Unit 441: Microstructure 441': First microstructure 441”: Second microstructure 442: Bearing concave surface 443: Gathering Area 444: Guiding Area 45: Surface microstructures 800: Cells

Claims

1. A microtrough structure for cell culture, suitable for performing said cell culture, comprising: a microtrough body defining an upward-opening micro-culture space; and a microstructural unit disposed in the microtrough body and located in the micro-culture space, the microstructural unit comprising a plurality of microstructures extending longitudinally and protruding into the rim of the micro-culture space, the microstructures cooperating to define a bearing concave surface with a gradually narrowing diameter from top to bottom, which can be used to carry said cells, the bearing concave surface having a centrally located aggregation area and an annular guiding area extending obliquely upward and outward from the periphery of the aggregation area, the microstructures being divided into a plurality of first microstructures cooperating to define the aggregation area and a plurality of second microstructures cooperating to define the guiding area, the first microstructures being adjacent and connected tubes, and the second microstructures being spaced-apart columns.

2. The microgroove structure for cell culture as described in claim 1, wherein, Each of the first microstructures is a hexagonal tube, and the first microstructures are adjacent to each other in a honeycomb pattern, with their top surfaces matching to define the aggregation area. Each of the second microstructures is a hexagonal prism, and the second microstructures are arranged at intervals in a honeycomb pattern, with their top surfaces matching to define the guiding area.

3. The microgroove structure for cell culture as described in claim 1, wherein, The cluster area is below the guide area.

4. The microgroove structure for cell culture as described in claim 2, wherein, The maximum diameter of each of the second microstructures is DO, and the center-to-center distance (P) between two adjacent second microstructures is 1.1DO to 3DO.

5. The microgroove structure for cell culture as described in claim 4, wherein, The maximum inner diameter of each of the first microstructures is Di, and the center-to-center distance (P) between two adjacent first microstructures is 1.1Di to 3Di.

6. The microgroove structure for cell culture as described in claim 5, wherein, The area of ​​this cluster ranges from 0.01 to 20 mm2.

7. The microgroove structure for cell culture as described in any one of claims 1 to 6, wherein, The microgroove has an annular opening wall that extends vertically upwards and an upward-facing bottom wall that extends downwards from the bottom edge of the opening wall. The microstructures of the microgroove structure extend vertically along the inner edge of the bottom wall.

8. The microgroove structure for cell culture as described in claim 7, wherein, These microstructures protrude vertically upwards from the inner edge of the bottom wall of the trench.

9. The microgroove structure for cell culture as described in claim 7, wherein, These microstructures protrude upwards perpendicularly to the inner edge of the bottom wall of the trench.

10. A cell culture container suitable for cell culture, comprising: a substrate; and a plurality of cell culture tank units arranged at intervals on the substrate, each cell culture tank unit including a main tank extending concavely downward from the top surface of the substrate and having an opening facing upward, and at least one microtrough structure disposed in the main tank for cell culture, each microtrough structure having, A microtrough is recessed into the corresponding main trough, defining an upward-facing micro-culture space. A microstructure unit is disposed in the microtrough and located within the micro-culture space. The microstructure unit includes multiple microstructures extending longitudinally along the inner edge of the micro-culture space. These microstructures cooperate to define a support concave surface with a gradually narrowing diameter from top to bottom, which can be used to support the cells. The support concave surface has a central aggregation area and an annular guiding area extending obliquely upward and outward from the periphery of the aggregation area. The microstructures are divided into multiple first microstructures that cooperate to define the aggregation area and multiple second microstructures that cooperate to define the guiding area. The first microstructures are adjacent and connected tubes, and the second microstructures are spaced-apart columns.

11. The cell culture container as claimed in claim 10, wherein, The cluster area is below the guide area.

12. The cell culture container as claimed in claim 10, wherein, Each of the first microstructures is a hexagonal tube, and the first microstructures are adjacent and connected in a honeycomb pattern, with their top surfaces matching to define the aggregation area. Each of the second microstructures is a hexagonal prism, and the second microstructures are arranged at intervals in a honeycomb pattern, with their top surfaces matching to define the guiding area.

13. The cell culture container as claimed in claim 12, wherein, The diameter of each of the second microstructures is DO, and the center-to-center distance (P) between two adjacent second microstructures is 1.1DO to 3DO.

14. The cell culture container as claimed in claim 12, wherein, The inner diameter of each of the first microstructures is Di, and the center-to-center distance (P) between two adjacent first microstructures is 1.1Di to 3Di.

15. The cell culture container as claimed in claim 10, wherein, The area of ​​this cluster ranges from 0.01 to 20 mm2.

16. The cell culture container as described in any one of claims 10 to 15, wherein, Each of the main tanks has a concave bottom edge, and each cell culture tank unit includes a plurality of microgroove structures arranged on the concave bottom edge of the main tank.

17. The cell culture container as claimed in claim 16, wherein, Each of the microgrooves has an annular opening wall that extends vertically downward from the corresponding concave bottom edge, and a groove bottom wall that extends downward from the bottom edge of the periphery of the opening wall and gradually narrows in diameter downward. The microstructures of each microgrooves extend vertically and are disposed on the inner edge of the groove bottom wall.

18. A cell culture container suitable for cell culture, comprising: a substrate; and a plurality of cell culture tank units arranged at intervals on the substrate, each cell culture tank unit including a main tank extending concavely downward from the top surface of the substrate and opening upward, and a plurality of microgroove structures arranged in the main tank for cell culture, each main tank having a concave bottom edge for the microgroove structures, each concave bottom edge defining a spacer structure separating the microgroove structures, each cell culture tank unit further including a plurality of surface microstructures protruding from the top surface of the spacer structure, the top edges of the surface microstructures being distributed in an upwardly arcuate pattern, each microgroove structure having... A microtrough is recessed into the corresponding main trough and defines an upward-facing micro-culture space. A microstructure unit is disposed in the microtrough and located in the micro-culture space. The microstructure unit includes multiple microstructures extending longitudinally in the vertical direction and disposed within the inner edge of the micro-culture space. These microstructures cooperate to define a bearing concave surface with a gradually narrowing diameter from top to bottom, which can be used to support the cells.

19. The cell culture container as claimed in claim 18, wherein, These surface microstructures are columnar extending vertically, and are spaced apart from each other.

20. The cell culture container as claimed in claim 18, wherein, These surface microstructures are tubular structures extending vertically, and these surface microstructures are connected to each other.

21. The cell culture container as claimed in claim 18, wherein, These microstructures protrude vertically upwards from the inner edge of the bottom wall of the groove.

22. The cell culture container as claimed in claim 18, wherein, These microstructures protrude obliquely upwards from the inner edge of the bottom wall of the groove.

23. The cell culture container as claimed in claim 18, wherein, The opening wall of each micro-groove is circular or polygonal, and the bottom wall of each micro-groove extends downward from the bottom edge of the opening wall in a conical shape. The bearing concave surface formed by the microstructures disposed on the bottom wall of each micro-groove is polygonal pyramidal, and the upward and downward inclined extension angle of the bearing concave surface can be between 10 degrees and 75 degrees.

24. The cell culture container as claimed in claim 23, wherein, The depth of each microgroove structure from the opening of the micro-culture space down to the lowest point of the bearing concave surface is 0.1 to 1.5 mm.

25. The cell culture container as claimed in claim 18, wherein, The opening wall of each micro-groove is in the shape of a circular ring or a polygonal ring. The bottom wall of each micro-groove has a narrow ring segment connected to the bottom edge of the opening wall and gradually narrowing downwards, and a bottom segment connected to the bottom edge of the narrow ring segment and having an arc-shaped opening facing upwards. The vertical inclination angle of the inner edge of the narrow ring segment is between 10 and 75 degrees.

26. The cell culture container as claimed in claim 25, wherein, The diameter of the bottom section of the trough is between 0.02 and 2.5 mm.

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