A basket type ultra-low adsorption three-dimensional cell culture device

Abstract

The utility model discloses a basket type ultralow adsorption three -dimensional cell culture device, including outer culture board, inner basket type culture board and upper seal cover, a plurality of rectangular array arrangement and top opening setting's inner culture hole cavities are fixed in the bottom of inner basket type culture board, the setting groove for placing inner basket type culture board is provided on outer culture board, and a plurality of outer culture hole cavities that are set up one by one with the inner culture hole cavities up and down are provided on the setting groove, the inner culture hole cavities are composed of the cylindrical barrel and U type filter basket that set up from top to bottom, meanwhile, the U type filter basket is PES microporous filter screen, and the surface is provided with the brush -like hydrophilic modification layer that grafts mPEG after plasma activation, and its advantage lies in having ultralow adsorption surface and efficient liquid exchange function, can the good protection cell ball of while, reduce cell adhesion, effectively overcome the cell ball loss, the problem such as complicated operation caused in the liquid exchange process of prior art.

Description

Technical Field

[0001] This invention belongs to the field of cell culture technology, and in particular relates to a basket-type ultra-low adsorption three-dimensional cell culture device. Background Technology

[0002] Three-dimensional cell culture, with its ability to better simulate the spatial structure of cells in vivo, cell-cell interactions, cell-matrix interactions, and biochemical gradients, provides a more physiologically relevant and predictable in vitro model. It has great potential in basic research, drug development, and regenerative medicine, and is particularly suitable for high-throughput drug screening, organoid culture, tumor model construction, stem cell research, tissue engineering, and toxicology testing.

[0003] Existing three-dimensional cell culture media changes require repeated centrifugation or aspiration, which can easily lead to mechanical damage and loss of cell spheres, resulting in a large loss of cells. Furthermore, the operation is cumbersome and difficult. Therefore, current technology lacks an integrated device that can both protect the cell sphere structure and simplify media changes. Utility Model Content

[0004] The purpose of this invention is to provide a basket-type ultra-low adsorption three-dimensional cell culture device that combines an ultra-low adsorption surface with a high-efficiency medium exchange function. This device can effectively protect cell spheres while reducing cell adhesion, thus overcoming the problems of cell sphere loss and cumbersome operation caused by existing technologies during medium exchange.

[0005] The technical solution of this utility model is: a basket-type ultra-low adsorption three-dimensional cell culture device, including an outer culture plate, an inner basket-type culture plate that is detachably disposed inside the outer culture plate, and an upper sealing cover disposed on the top of the outer culture plate. Several inner culture cavities arranged in a rectangular array and open at the top are fixed at the bottom of the inner basket-type culture plate. The outer culture plate is provided with a placement groove for placing the inner basket-type culture plate, and several outer culture cavities are provided on the placement groove, which correspond one-to-one with the inner culture cavities. The inner culture cavity is composed of a cylindrical body and a U-shaped filter basket arranged from top to bottom. The U-shaped filter basket is a PES microporous filter screen, and a brush-shaped hydrophilic modification layer formed by grafting mPEG after plasma activation is provided on its surface.

[0006] As a preferred technical solution, two symmetrically arranged locking components for fixing the inner basket-type culture plate are provided on the front and rear sides of the placement groove. The locking components include a fixing strip and a movable push block. At the same time, a shrinkage groove corresponding to the fixing strip is provided on the inner side wall of the placement groove, and a push groove communicating with the inner end of the shrinkage groove is opened on the upper surface of the outer culture plate. The movable push block is disposed inside the push groove, and its bottom is fixedly connected to the inner end of the fixing strip.

[0007] As a preferred technical solution, the thickness of the brush-like hydrophilic modified layer is 8~12 nm, and its grafting density is 0.5~1.0 chains / nm. 2 The static water contact angle is ≤15°.

[0008] As a preferred technical solution, the PES microporous filter screen is provided with a plurality of trapezoidal micropores arranged radially with the bottom center as the center point. The pore size of the trapezoidal micropores is 80~300um and increases in a stepwise manner from the center point outward, and the pore density is 15~20 pores / mm. 2 .

[0009] As a preferred technical solution, two symmetrically arranged arc-shaped notches are provided on both sides of the placement groove to clamp the inner basket-type culture plate.

[0010] As a preferred technical solution, the number of the inner culture cavities and the outer culture cavities are matched, and the height of the cavities is the same, with a center-to-center distance deviation of ≤0.1mm.

[0011] As a preferred technical solution, the inner wall of the external culture cavity is covered with an ultra-low adsorption coating, which is a medical-grade hydrophilic polymer.

[0012] As a preferred technical solution, the bottom of the U-shaped filter basket has an inclination angle of 45° to 60°.

[0013] As a preferred technical solution, the inner side of the upper sealing cover is provided with a plurality of grooves for the retaining strip and the movable push block, which are respectively provided in a one-to-one correspondence with the shape of the retaining strip and the movable push block.

[0014] The advantages of this utility model are: This invention relates to a basket-type ultra-low adsorption three-dimensional cell culture device. By providing a brush-shaped hydrophilic modified layer formed by grafting mPEG after plasma activation on the surface of a PES microporous filter, sufficient space repulsion layer thickness is provided to effectively repel proteins and cells and prevent them from contacting the bottom of the PES microporous filter, thereby achieving ultra-low adsorption. This makes the liquid replacement operation extremely gentle and minimizes cell spheroid damage and experimental errors caused by cell adhesion. This utility model's basket-type ultra-low adsorption three-dimensional cell culture device, through the separable design of the outer culture plate and the inner basket-type culture plate, can achieve vertical lifting for medium exchange. This not only avoids manual aspiration of culture medium one by one, simplifying the operation steps and greatly shortening the medium exchange time, but also avoids the pipette tip touching the cell ball, reducing cell ball loss. This invention features trapezoidal micropores on a PES microporous filter mesh that are radially distributed and increase in size stepwise from the center outwards. This design allows for the blocking of cell spheres in the central bottom area and rapid filtration in the side edge areas, significantly reducing the liquid change time. Attached Figure Description

[0015] To more clearly illustrate the technical solutions of the embodiments of this utility model, the drawings used in the following description of the embodiments will be briefly introduced. The drawings described below are only some embodiments of this utility model. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0016] The present invention will be further described below with reference to the accompanying drawings and embodiments: Figure 1 This is an assembly diagram of the overall structure of this utility model; Figure 2 This is a partial enlarged view of the inner basket-type culture plate of this utility model; Figure 3 This is a schematic diagram of the external culture plate structure of this utility model; Figure 4 This is a bottom view of the overall structure of this utility model; Among them: 1 outer culture plate, 11 placement groove, 12 outer culture cavity, 13 fixing strip, 14 movable push block, 15 arc-shaped notch; 2. Inner basket-type culture plate, 21. Inner culture well cavity, 211. Cylindrical body, 212. U-shaped filter basket, 213. Trapezoidal micropores; 3. Upper sealing cover, 31. Clip groove, 32. Push block groove. Detailed Implementation

[0017] The above solution will be further described below with reference to specific embodiments. It should be understood that these embodiments are for illustrating the present invention and are not intended to limit the scope of the present invention. The implementation conditions used in the embodiments can be further adjusted according to the conditions of specific manufacturers, and the implementation conditions not specified are usually the conditions in conventional experiments.

[0018] Example: This invention is suitable for high-throughput three-dimensional culture (6 / 12 / 24 / 96 wells), compatible with microscope observation, with a light transmittance >90%, and can be sterilized at high temperature (121℃, 20 min).

[0019] The external culture plate 1 is mainly used for holding the culture medium. In some embodiments, the number of wells in the external culture plate 1 can be set to 6-well plate, 12-well plate, 24-well plate, 96-well plate, etc. For this embodiment... Figure 3 As shown, the external culture plate 1 is set as a 24-well plate.

[0020] The inner basket-type culture plate 2 is mainly used for support. In some embodiments, the number of wells in the inner basket-type culture plate 2 can be set to 6-well plate, 12-well plate, 24-well plate, 96-well plate, etc. For this embodiment... Figure 1 As shown, the inner basket-type culture plate 2 is set as a 24-well plate. It should be understood that no matter what kind of well plate the outer culture plate 1 is set as, the number of inner culture wells 21 and outer culture wells 12 are matched with each other, and the height of their wells is the same, with a center distance deviation of ≤0.1mm.

[0021] Reference Figures 1 to 4 As shown, a basket-type ultra-low adsorption three-dimensional cell culture device includes an outer culture plate 1, an inner basket-type culture plate 2 detachably disposed inside the outer culture plate 1, and an upper sealing cover 3 disposed on the top of the outer culture plate 1; 24 inner culture cavities 21 arranged in a rectangular array and with open tops are fixed at the bottom of the inner basket-type culture plate 1; a placement groove 11 for placing the inner basket-type culture plate 2 is provided on the outer culture plate 1, and 24 outer culture cavities 12 are provided on the placement groove 11 corresponding one-to-one with the inner culture cavities 21; the inner culture cavity 21 is composed of a cylindrical body 211 arranged from top to bottom and a U-shaped filter basket 212, and the U-shaped filter basket 212 is a PES microporous filter, and a brush-shaped hydrophilic modification layer formed by grafting mPEG after plasma activation is provided on its surface.

[0022] In this embodiment, the thickness of the brush-like hydrophilic modified layer is 8~12 nm, and its grafting density is 0.5~1.0 chains / nm. 2 (Grafted molecular weight 5 kDa), static water contact angle ≤15°; by setting a brush-like hydrophilic modified layer (bioinert surface) of 8~12 nm formed by plasma-activated grafted mPEG on the surface of the PES microporous filter (mechanical framework), sufficient space repulsion layer thickness is provided to effectively repel proteins and cells, preventing them from contacting the bottom of the PES microporous filter, with a grafting density of 0.5~1.0 chains / nm. 2 It forms a dense, non-porous molecular barrier with extremely low protein adsorption, preventing cells from anchoring and eliminating the possibility of biomolecules penetrating. The static water contact angle is ≤15°, forming a strong hydration layer. Through the "water shield" effect, it physically blocks biomolecules. The hydrophilic surface greatly reduces hydrophobic interactions and electrostatic adsorption, thereby achieving ultra-low adsorption. This makes the liquid changing operation extremely gentle and minimizes cell spheroid damage and experimental errors caused by cell adhesion.

[0023] Reference Figure 3 As shown, in this embodiment, two symmetrically arranged locking components for fixing the inner basket-type culture plate 2 are provided on the front and rear sides of the placement groove 11. The locking components include a fixing strip 13 and a movable push block 14. At the same time, a shrinkage groove corresponding to the fixing strip 13 is provided on the inner side wall of the placement groove 11, and a push groove communicating with the inner end of the shrinkage groove is opened on the upper surface of the outer culture plate 1. The movable push block 14 is disposed inside the push groove, and its bottom is fixedly connected to the inner end of the fixing strip 13, which can realize the fixing or release of the inner basket-type culture plate 2. Meanwhile, a strip groove 31 and a push block groove 32 corresponding to the shapes of the fixing strip 13 and the movable push block 14 are provided on the inner side of the upper sealing cover 3.

[0024] Reference Figure 2 As shown, in this embodiment, a plurality of trapezoidal micropores 213 are arranged radially with the bottom center as the center point on the PES microporous filter screen (the upper base width / lower base width of the trapezoidal micropore 213 is 1.5:1). The pore diameter of the trapezoidal micropores 213 is 80~300um and increases in a stepwise manner from the center point outward; wherein the pore diameter of the central area (circular area with a diameter of 5 mm) is 80~100μm, and the pore density is 20 pores / mm. 2 The transition zone (ring width 2 mm) has a pore size of 100~150 μm and a pore density of 18 pores / mm². 2 The pore size in the edge region (outermost 3 mm) is 200~300 μm, and the pore density is 15 pores / mm². 2 It can block cell spheres (100um) in the bottom central area and quickly filter the liquid in the two side edge areas (flow rate ≥0.5 mL / s), which greatly shortens the medium change time. It not only avoids manually aspirating the culture medium one by one, simplifying the operation steps and greatly shortening the medium change time to achieve rapid medium change (the medium change time for 24-well plates is <1min), but also avoids the pipette tip touching the cell spheres, reducing cell sphere loss.

[0025] In this embodiment, two symmetrical arc-shaped notches 15 (3-5mm) are provided on both sides of the placement groove 11 to clamp the inner basket-type culture plate 2, forming a sterile operation clamping space.

[0026] In this embodiment, the inner wall of the external culture cavity 12 is covered with an ultra-low adsorption coating, which is a medical-grade hydrophilic polymer that can achieve ultra-low adsorption.

[0027] In this embodiment, the bottom of the U-shaped filter basket 212 is inclined at a 60° angle, which allows the cell spheres to slide towards the center area under gravity, and the liquid shear force is relatively small.

[0028] The medium replacement operation in this embodiment is simple: When performing the medium replacement operation, first open the upper sealing cover 3, push the movable push block 14 on the outer culture plate 1 outward, so that the two fixing strips 13 retract and release the inner basket-type culture plate 2. Use two fingers or tweezers to lift the inner basket-type culture plate 2 directly from the arc-shaped notch 15. When the inner basket-type culture plate 2 is lifted and all the liquid leaks into the outer culture plate 1, the culture medium of the outer culture plate 1 can be poured out all at once. Then put the inner basket-type culture plate 2 back and fix it again. After adding new culture medium, the next operation can be carried out.

[0029] The above embodiments are only for illustrating the technical concept and features of this utility model, and are intended to enable those skilled in the art to understand the content of this utility model and implement it accordingly. They should not be construed as limiting the scope of protection of this utility model. All equivalent changes or modifications made in accordance with the spirit and essence of this utility model should be included within the scope of protection of this utility model.

Claims

1. A basket-type ultra-low adsorption three-dimensional cell culture device, characterized in that, It includes an outer culture plate, an inner basket-type culture plate that is detachably disposed inside the outer culture plate, and an upper sealing cover disposed on the top of the outer culture plate; Several inner culture cavities arranged in a rectangular array and open at the top are fixed at the bottom of the inner basket-type culture plate. The outer culture plate is provided with a placement groove for placing the inner basket-type culture plate, and several outer culture cavities are provided on the placement groove, which correspond one-to-one with the inner culture cavities. The inner culture cavity is composed of a cylindrical body and a U-shaped filter basket arranged from top to bottom. The U-shaped filter basket is a PES microporous filter screen, and a brush-shaped hydrophilic modification layer formed by grafting mPEG after plasma activation is provided on its surface.

2. The basket-type ultra-low adsorption three-dimensional cell culture device according to claim 1, characterized in that, Two symmetrically arranged locking assemblies for fixing the inner basket-type culture plate are provided on the front and rear sides of the placement groove. The locking assembly includes a fixed locking strip and a movable push block. At the same time, a shrinkage groove corresponding to the fixed locking strip is provided on the inner side wall of the placement groove, and a push groove communicating with the inner end of the shrinkage groove is opened on the upper surface of the outer culture plate. The movable push block is disposed inside the push groove, and its bottom is fixedly connected to the inner end of the fixed locking strip.

3. The basket-type ultra-low adsorption three-dimensional cell culture device according to claim 1, characterized in that, The thickness of the brush-like hydrophilic modified layer is 8~12 nm, and its grafting density is 0.5~1.0 chains / nm. 2 The static water contact angle is ≤15°.

4. The basket-type ultra-low adsorption three-dimensional cell culture device according to claim 1, characterized in that, The PES microporous filter screen has a plurality of trapezoidal micropores arranged radially around the bottom center. The pore size of the trapezoidal micropores is 80~300um and increases in a stepwise manner from the center outwards, and the pore density is 15~20 pores / mm. 2 .

5. The basket-type ultra-low adsorption three-dimensional cell culture device according to claim 1, characterized in that, Two symmetrically arranged arc-shaped notches are provided on both sides of the placement groove to clamp the inner basket-type culture plate.

6. The basket-type ultra-low adsorption three-dimensional cell culture device according to claim 1, characterized in that, The number of inner culture cavities and the number of outer culture cavities are matched, and the height of the cavities is the same, with a center-to-center distance deviation of ≤0.1mm.

7. The basket-type ultra-low adsorption three-dimensional cell culture device according to claim 1, characterized in that, The inner wall of the external culture cavity is covered with an ultra-low adsorption coating, which is a medical-grade hydrophilic polymer.

8. The basket-type ultra-low adsorption three-dimensional cell culture device according to claim 1, characterized in that, The bottom of the U-shaped filter basket has an inclination angle of 45° to 60°.

9. The basket-type ultra-low adsorption three-dimensional cell culture device according to claim 2, characterized in that, The inner side of the upper sealing cover is provided with a plurality of grooves for the retaining strip and the movable push block, which are respectively provided in a one-to-one correspondence with the shape of the retaining strip and the movable push block.

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