Non-contact cell co-culture device

Abstract

The utility model discloses a non-contact type cell co-culture device, which relates to the technical field of cell culture and comprises an upper-layer frame body and a lower-layer culture dish for arranging lower-layer cells, upper-layer cells planted on a climbing slide are arranged on the upper-layer frame body, the upper-layer frame body is arranged in the lower-layer culture dish in a lap joint mode, and the upper-layer cells and the lower-layer cells are arranged in a non-contact mode. The upper layer frame body comprises an upper surrounding support, a lower surrounding support and an inner side flange, the upper end of the lower surrounding support is connected with the lower end of the upper surrounding support, the lower end of the lower surrounding support is connected with the inner side flange, and an annular frame body with the hollow middle is formed; the inner side flange is arranged on the inner side edge of the bottom of the annular frame body; according to the cell co-culture device, double-layer cells can be effectively observed, and the double-layer cells can be efficiently utilized.

Description

Technical Field

[0001] The utility model relates to the technical field of cell culture, in particular to a non-contact cell co-culture device. Background Art

[0002] Studying intercellular interactions is crucial to understanding the basic processes of life activities and the mechanisms of disease occurrence. For example, the communication between neurons and glial cells affects the maturation and myelination of neurons, the mutual influence between cardiomyocytes and fibroblasts participates in heart development, and the interaction between dendritic cells and T lymphocytes participates in the antigen presentation process in the immune response. However, single cell culture can no longer meet the needs of scientific research. Cell co-culture is an important means to study intercellular interactions.

[0003] At present, co-culture technology has developed from direct contact to indirect contact, and from two-dimensional to three-dimensional culture. Traditional direct contact co-culture is to mix different types of cells in the same culture system. Although it can make the cells directly contact and interact with each other, the different types of cells cannot be completely separated, which is not conducive to subsequent separation and analysis and observation. Indirect contact co-culture is to inoculate cells into different carriers or positions separately and place them in the same culture environment. Through paracrine cytokine interaction, it is easy to observe and does not affect subsequent detection. However, there are problems such as complex model preparation process and cumbersome operation.

[0004] Commonly used non-contact co-culture devices on the market, such as those using a combination of Matrigel and Transwell culture chambers, are expensive, difficult to operate, and incapable of co-culturing more than two adherent cell types. Furthermore, in practice, the porous membranes commonly used in Transwell culture chambers have limitations in pore size selection, unstable permeability, reduced experimental reproducibility, poor mechanical strength and susceptibility to breakage, and poor optical transparency, making it difficult to observe the cell layer on the membrane and hindering subsequent use and processing, all of which hinder the development of cell co-culture research. Utility Model Content

[0005] Based on the technical problems existing in the background technology, the utility model proposes a non-contact cell co-culture device, which can efficiently utilize double-layer cells and realize effective observation of double-layer cells.

[0006] The utility model proposes a non-contact cell co-culture device, comprising an upper frame and a lower culture dish for setting lower cells. The upper cells seeded on a slide are set on the upper frame, and the upper frame is overlapped and set in the lower culture dish. The upper cells and the lower cells are set in non-contact, and the upper cells and the lower cells achieve co-culture through the interaction of paracrine cytokines.

[0007] Furthermore, the upper frame includes an upper bracket, a lower bracket and an inner flange. The upper end of the lower bracket is connected to the lower end of the upper bracket, and the lower end is connected to the inner flange to form a hollow annular frame in the middle. The inner flange is arranged on the inner edge of the bottom of the annular frame.

[0008] Furthermore, the annular diameter of the upper end of the upper bracket is larger than the annular diameter of the lower end, and the annular diameter of the lower bracket is equal to the annular diameter of the lower end of the upper bracket.

[0009] Furthermore, bracket gaps are provided at intervals on the lower bracket.

[0010] Furthermore, a limiting bracket is left between the bracket notch and the inner flange of the lower bracket, and the height of the limiting bracket is greater than or equal to the thickness of the climbing plate.

[0011] Furthermore, the bracket notch is arranged in a U shape on the lower bracket.

[0012] Furthermore, the upper frame body is provided with a side notch along the axial direction, and the side notch passes through the upper surrounding bracket, the lower surrounding bracket, and the inner flange.

[0013] Furthermore, the width of the side notch at the upper end of the upper bracket is greater than the width of the side notch at the lower end of the upper bracket.

[0014] Furthermore, outer flanges are provided at intervals on the outer periphery of the upper end of the upper bracket, and the outer flanges overlap with the upper end opening of the lower culture dish.

[0015] The upper end of the upper bracket is bent outward to form a bent portion, and the outer flange is connected to the bent portion.

[0016] The advantages of the non-contact cell co-culture device provided by the utility model are: it replaces the Transwell culture chamber made of porous membrane material, has low cost, flexible operation and convenience, can efficiently utilize double-layer cells, realizes effective observation of double-layer cells, and is conducive to interaction research and cell separation and purification. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 It is a structural diagram of the utility model;

[0018] Figure 2 This is a schematic diagram of the structure of the upper frame equipped with climbing plates;

[0019] Figure 3 Schematic diagram of the structure of the upper frame;

[0020] Among them, 1-upper frame, 2-lower culture dish, 3-lower cells, 4-upper cells, 5-tweezers, 6-climbing slide, 11-upper bracket, 12-lower bracket, 13-inner flange, 14-bracket notch, 15-limiting bracket, 16-side notch, 17-outer flange, 18-bending part, 19-positioning notch. DETAILED DESCRIPTION

[0021] The following describes the technical solution of the present invention in detail through specific embodiments. Many specific details are set forth in the following description to facilitate a full understanding of the present invention. However, the present invention can be implemented in many other ways than those described herein, and those skilled in the art can make similar improvements without violating the scope of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0022] like Figures 1 to 3 As shown, the utility model proposes a non-contact cell co-culture device, including an upper frame 1 and a lower culture dish 2 with lower cells 3 arranged thereon, the upper cells 4 seeded on the climbing sheet 6 are arranged on the upper frame 1, the upper frame 1 is overlapped and arranged in the lower culture dish 2, the upper cells 4 and the lower cells 3 are arranged in a non-contact manner, and the upper cells 4 and the lower cells 3 achieve co-culture through the interaction of paracrine cytokines.

[0023] In this embodiment, a conventional glass climbing sheet 6 or a plastic climbing sheet 6 of appropriate size is used instead of the commonly used porous membrane material. The upper cells 4 are planted on the climbing sheet 6. The glass or plastic climbing sheet 6 can be placed on the upper frame 1 using tools such as tweezers, and the upper cell surface 4 can be selected to face up or down. The effective components secreted by the upper cells can exchange substances with the lower cells, and the culture process can be directly observed under a microscope.

[0024] In one embodiment, the upper frame 1 includes an upper bracket 11, a lower bracket 12 and an inner flange 13. The upper end of the lower bracket 12 is connected to the lower end of the upper bracket 11, and the lower end is connected to the inner flange 13 to form a hollow annular frame in the middle. The inner flange 13 is arranged on the inner edge of the bottom of the annular frame.

[0025] The inner flange 13 is used to place the upper cells 4. The upper bracket 11 and the lower bracket 12 can be formed as a whole or separately and then connected. In order to facilitate the placement of the upper cells 4, the upper frame 1 is provided with a side notch 16 in the axial direction. The side notch 16 passes through the upper bracket 11, the lower bracket 12, and the inner flange 13. The climbing sheet 6 planted with the upper cells 4 is clamped by the tweezers 5 and moved from the upper end to the lower end of the upper frame 1. During the movement, the tweezers 5 move along the side notch 16, and the climbing sheet 6 moves in the annular frame until the climbing sheet 6 overlaps the inner flange 13.

[0026] When the climbing slide 6 is moved up and down by the tweezers 5, in order to avoid the friction problem between the climbing slide 6 and the upper frame 1 when it moves up and down, the upper end annular diameter of the upper bracket 11 is set to be larger than the lower end annular diameter, and the annular diameter of the lower bracket 12 is equal to the lower end annular diameter of the upper bracket 11, thereby forming a funnel-like structure, which is not only conducive to the clamping of the upper frame 1, but also allows the climbing slide 6 to quickly pass through the funnel-like structure and overlap on the inner flange 13; in addition, this funnel-like structure setting allows the annular diameter of the lower bracket 12 to be designed according to the direct size of a conventional glass slide, so that glass slides of different specifications can be adapted by adjusting the diameter of the lower bracket 12.

[0027] In addition, the bracket notch 14 and the side notch 16 allow the secretory factors secreted by the upper layer cells 4 and the lower layer cells 3 to be effectively circulated through the culture medium, eliminating the need to select the transwell chamber membrane material and pore size based on the experimental purpose and cell type. At the same time, the selection of a transwell chamber requires consideration of multiple factors, such as the cell, membrane material properties, membrane pore size, and cell adhesion properties. These factors limit the selection of a transwell chamber and the observation of the cells in the upper layer. In this embodiment, a glass or transparent plastic slide 6 is used, which eliminates the need to consider these numerous factors. The slide is hard and not easily damaged, and has good light transmittance, making it easy to observe the state of the upper layer cells under a microscope. Pretreatment such as appropriate embedding can also be performed based on actual conditions.

[0028] Finally, the design of the side notch 16, when used with clamping tools such as tweezers, can flexibly place and take the upper cells 4 seeded on the slide 6, so as to realize complex experimental processing such as replacement of co-cultured cells and subsequent labeling and imaging of the upper cells 4, thereby solving the problem that the Transwell membrane material is difficult to subsequently process and utilize.

[0029] In this embodiment, in order to achieve the overlapping setting between the upper frame 1 and the lower culture dish 2, the outer flange 17 is arranged at intervals on the outer periphery of the upper end of the upper bracket 11, and the outer flange 17 overlaps the upper end opening of the lower culture dish 2; since the outer flanges 17 are arranged at intervals, a locking notch 19 is provided between adjacent outer flanges 17. The setting of the locking notch 19 makes it convenient to use tweezers 5 or other clamping mechanisms to clamp the upper frame 1, so as to achieve the use of the upper frame 1.

[0030] To increase the stability of the connection between outer flange 17 and the upper end of upper bracket 11, the upper end of upper bracket 11 is bent outward to form a bent portion 18, and outer flange 17 is connected to bent portion 18. The provision of bent portion 18 increases the fixing area between outer flange 17 and upper bracket 11, thereby increasing the structural stability of outer flange 17.

[0031] In order to better realize the co-culture of the upper cells 4 and the lower cells 3 through the interaction of paracrine cytokines, the lower bracket 12 is provided with bracket gaps 14 at intervals. From the appearance of the lower bracket 12, the bracket gaps 14 can be square, circular, diamond-shaped or the like; as for the opening height of the bracket gap 14, it can pass through the upper end of the lower bracket 12 to form a U shape, or it can leave a certain height at the upper end of the lower bracket 12, so as to realize the circulation of paracrine cytokines between the upper cells 4 and the lower cells 3.

[0032] In addition, in order to prevent the upper cells 4 from falling into the lower culture dish 2 through the edge of the climbing sheet 6, the bracket gap 14 does not pass through the lower end of the lower bracket 12, that is, in appearance, the lower bracket 12 leaves a limiting bracket 15 between the bracket gap 14 and the inner flange 13, and the height of the limiting bracket 15 is greater than or equal to the thickness of the climbing sheet 6, thereby limiting the edge of the climbing sheet 6 to prevent the upper cells 4 from falling directly. At the same time, due to the existence of the bracket gap 14, paracrine cytokines can flow through the bracket gap 14, so that the upper cells 4 and the lower cells 3 can indirectly interact with each other.

[0033] The most commonly used membrane materials for existing Transwell chambers are polycarbonate (PC) and polyester (PET). PC membrane is a translucent membrane with poor light transmittance, making it difficult to observe cells. PC membrane has good physical and mechanical properties, but poor chemical stability and is easily hydrolyzed and cracked in water for a long time. Although PET has good light transmittance, the actual observation effect under a microscope is still average. It usually needs to be combined with a high-performance camera or staining for observation and imaging. In addition, compared with PC membrane, it has weaker impact resistance and is easily damaged by impact. In this embodiment, an upper frame 1 is provided to accommodate a climbing slide 6 seeded with upper cells 4. The climbing slide 6 can be an existing glass climbing slide or plastic climbing slide, etc., replacing the Transwell culture chamber made of porous membrane material. It has low cost, flexible operation and easy use, can efficiently utilize double-layer cells, realize effective observation of double-layer cells, and is conducive to interaction research and cell separation and purification. It is a non-contact cell co-culture device.

[0034] The above is only a preferred specific implementation method of the present invention, but the protection scope of the present invention is not limited to this. Any technician familiar with the technical field within the technical scope disclosed by the present invention can make equivalent replacements or changes based on the technical solution and utility model concept of the present invention, which should be covered by the protection scope of the present invention.

Claims

1. A non-contact cell co-culture device, characterized in that: The invention comprises an upper frame (1) and a lower culture dish (2) in which lower cells (3) are arranged. Upper cells (4) planted on a slide (6) are arranged on the upper frame (1). The upper frame (1) is overlapped and arranged in the lower culture dish (2). The upper cells (4) and the lower cells (3) are arranged in a non-contact manner. The upper cells (4) and the lower cells (3) achieve co-culture through the interaction of paracrine cytokines. The upper frame (1) comprises an upper frame bracket (11), a lower frame bracket (12) and an inner flange (13), wherein the upper end of the lower frame bracket (12) is connected to the lower end of the upper frame bracket (11), and the lower end is connected to the inner flange (13), forming an annular frame with a hollow center, and the inner flange (13) is arranged on the inner edge of the bottom of the annular frame.

2. The non-contact cell co-culture device according to claim 1, characterized in that: The annular diameter of the upper end of the upper bracket (11) is greater than the annular diameter of the lower end, and the annular diameter of the lower bracket (12) is equal to the annular diameter of the lower end of the upper bracket (11).

3. The non-contact cell co-culture device according to claim 1, characterized in that: Bracket notches (14) are provided at intervals on the lower bracket (12).

4. The non-contact cell co-culture device according to claim 3, characterized in that: The lower bracket (12) leaves a limiting bracket (15) between the bracket notch (14) and the inner flange (13), and the height of the limiting bracket (15) is greater than or equal to the thickness of the climbing plate (6).

5. The non-contact cell co-culture device according to claim 3, characterized in that: The bracket notch (14) is arranged in a U-shape on the lower bracket (12).

6. The non-contact cell co-culture device according to claim 1, characterized in that: The upper frame (1) is provided with a side notch (16) along the axial direction, and the side notch (16) is provided through the upper surrounding bracket (11), the lower surrounding bracket (12), and the inner flange (13).

7. The non-contact cell co-culture device according to claim 6, characterized in that: The width of the side notch (16) at the upper end of the upper bracket (11) is greater than the width of the side notch (16) at the lower end of the upper bracket (11).

8. The non-contact cell co-culture device according to claim 1, characterized in that: The outer periphery of the upper end of the upper bracket (11) is provided with outer flanges (17) at intervals, and the outer flanges (17) overlap the upper end opening of the lower culture dish (2).

9. The non-contact cell co-culture device according to claim 8, characterized in that: The upper end of the upper bracket (11) is bent outward to form a bent portion (18), and the outer flange (17) is connected to the bent portion (18).

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