Culture device for cell migration experiment
By introducing a membrane strip with grid scale lines into the cell culture device, the problems of inaccurate position and difficult positioning in the cell scratch experiment are solved, a simple and low-cost cell migration experiment is realized, and the accuracy and efficiency of the experiment are improved.
Patent Information
- Application Number
- CN202422707955.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-07
- Publication Date
- 2025-09-19
- Estimated Expiration
- 2034-11-07
AI Technical Summary
In existing cell scratch experiments, the scratch position is inaccurate and lacks positioning markers, resulting in inaccurate experimental results. In addition, existing dedicated culture dishes are expensive and cumbersome to operate.
A culture device was designed, comprising a base plate, a well plate and a membrane strip. The membrane strip is provided with grid scale lines, and the bottom of the culture wells is also provided with scale lines. The membrane strip is made of fibrinogen, which is easy to peel off and non-cytotoxic. There are tiny gaps between the membrane strips, providing regular scratches and positioning marks.
It achieves the formation of regular scratches, simplifies operations, improves experimental efficiency, reduces costs, enables non-destructive observation of cell migration, and facilitates positioning and data recording.
Smart Images
Figure CN223357660U_ABST
Abstract
Description
Technical Field
[0001] The utility model mainly relates to the technical field of cell experiment equipment, in particular to a culture device for cell migration experiments. Background Art
[0002] Testing cell migration is a commonly used experimental method in cell biology. The simplest method is the scratch wound test, which is widely used to determine the strength of cell migration. The main steps of this experiment are as follows: First, a monolayer of adherent cells of different types or that have undergone different treatments is seeded into a suitable culture dish until the cell confluence is ≥90%. A micropipette tip or other hard object is used to scratch the central area where the cells are growing, removing the cells in the central area. The cells are then cultured for the set experimental time (which can be different time points). The cell culture plate is removed and photographed under a microscope to obtain a sequence of images at multiple time points. The width of the scratch wound is then measured regularly to obtain the cell migration experimental data.
[0003] In the prior art, conventional cell scratch experiments have the following shortcomings: 1. The scratch experiment needs to ensure the accuracy of the scratch position. If the scratches are not parallel, the number of cells on both sides of the scratch will be different, which will affect the speed of cell migration to the middle of the scratch and reduce the accuracy of the experimental results. The conventional method is to purchase a gun tip for marking, and there is no positioning mark on the culture dish, which makes it almost impossible to guarantee that multiple parallel and equidistant scratches can be drawn. 2. The scratch experiment requires observing cells in the same field of view at multiple time points. However, there are no suitable marks on the culture dish, and it often takes a long time to find the field of view position of the last observation during the next observation, and it is impossible to measure the distance the cells move. There are no scale marks on the culture dish, which makes it impossible to intuitively measure the distance the cells move and then compare the cell migration ability.
[0004] While some specialized cell culture dishes for scratch experiments can partially address these challenges, these specialized dishes are often complex, expensive, and inconvenient to use. Therefore, there is an urgent need for a simple, cost-effective experimental device for testing cell migration ability that can be easily operated, provides regular scratches, and is inexpensive. However, no such device has been reported. Utility Model Content
[0005] Based on this, the purpose of the present invention is to provide a culture device for cell migration experiments to solve the technical problems raised in the above background technology.
[0006] To achieve the above objectives, the present invention provides the following technical solutions:
[0007] A culture device for cell migration experiments includes a base plate and a well plate on top of the base plate. The well plate has multiple culture wells distributed in a rectangular array on the top of the well plate. The bottom of the culture wells is provided with multiple membrane strips. The top of the membrane strips is provided with grid scale lines. The bottom of the culture wells and the bottom of the membrane strips are also provided with grid scale lines.
[0008] Preferably, tearing areas are provided at both ends of the film strip.
[0009] Preferably, the multiple membrane strips in the culture wells are arranged in parallel, and there are small intervals between adjacent membrane strips.
[0010] Preferably, the surface of the membrane strip is made of fibrinogen.
[0011] In summary, this technical solution has the following beneficial effects:
[0012] The utility model can obtain scratches with regular shapes, and the grid scale lines on the bottom of the plate will not cause damage to cells; the membrane strips are made of cell culture grade materials and will not float after being immersed in culture medium for 5 days; the operation is simple, which is conducive to batch operation in experiments and improves experimental efficiency.
[0013] In addition, it has a simple structure, is easy to produce, and has low cost; it is sterile and independently packaged, easy to use, and can be used at any time. BRIEF DESCRIPTION OF THE DRAWINGS
[0014] Figure 1 This is the axonometric drawing of the overall structure of the utility model;
[0015] Figure 2 This is a plan view of the structure of the culture wells and membrane strips of the present invention;
[0016] Figure 3 This is a partial cross-sectional view of the culture wells and membrane strips of the present invention;
[0017] Figure 4 This is an axonometric diagram of the culture well positions and grid scale line structure of the present invention.
[0018] Description of the accompanying drawings: 11, bottom plate; 12, well plate; 13, culture well position; 14, membrane strip; 15, grid scale line; 141, tearing area. DETAILED DESCRIPTION
[0019] The technical solutions in the embodiments of the present invention will be described clearly and completely below with reference to the accompanying drawings in the embodiments of the present invention.
[0020] like Figures 1 to 4As shown, a culture device for cell migration experiments is a device for scratch experiments, and also includes culture dishes, cell culture plates with various culture well specifications, including 24-well, 12-well, 6-well, etc.; it includes a base plate 11, a well plate 12 on top of the base plate 11, and a plurality of culture well positions 13 distributed in a rectangular array on the top of the well plate 12. A plurality of membrane strips 14 are provided at the bottom of the culture well positions 13, and tear areas 141 are provided at both ends of the membrane strips 14; the plurality of membrane strips 14 in the culture well positions 13 are arranged in parallel, and there is a small gap between adjacent membrane strips 14; the surface of the membrane strips 14 is made of fibrinogen material; the membrane strips 14 are non-cytotoxic, and no residue remains on the top of the bottom plate 11 after the membrane strips 14 are removed, which does not affect cell growth. During the scratch experiment, 1 to 5 membrane strips 14 can be removed as needed to obtain scratches of uniform size and specifications, which is convenient and fast, and is convenient for counting the cell movement position and calculating the relative scratch area.
[0021] The top of the membrane strip 14 is provided with grid scale lines 15, and the bottom of the culture well 13 and the bottom of the membrane strip 14 are also provided with grid scale lines 15. This facilitates observation and recording of cells, as well as secondary locating of cell positions.
[0022] It should be noted that, in this embodiment, the experimental steps are as follows:
[0023] 1. Cell preparation: Resuscitated adherent cells were cultured in a dedicated culture medium at 37°C and 5% CO2. When the cells reached 90% confluency, they were digested and counted.
[0024] 2. Seeding plate: According to the grouping seeding plate, take one 6-well standard plate designed by the utility model, add 1 ml of culture medium to each culture well 13 and rinse it once; the inoculation cell density is about 7.5x105 cells / well, and be sure to spread it evenly.
[0025] 3. Scratch: After the cells have grown fully, use tweezers to gently peel off two membrane strips 14 through the tearing area 141 at either end of the membrane strip 14, record the data of the grid scale line 15, and add PBS buffer to rinse the cells one or two times.
[0026] 4. Add liquid: Add drug-added culture medium or serum-free culture medium according to the group.
[0027] 5. Photographing: After scratching and adding solution, take microscopic photographs at different magnifications to serve as a 0-hour control. Incubate the cells in a 37°C, 5% CO2 incubator. Remove the cells at the desired time points, observe the scratch width at the same location under a microscope, take photos, and record the migration distance based on the 15-degree grid mark.
[0028] 6. Data Analysis: One method is to count the distance of cell migration, and the other is to count the area of the trace. Both methods can be analyzed using the data recorded by the grid scale line 15.
[0029] The working principle of this utility model is:
[0030] Taking a 6-well plate as an example, the bottom center of the well plate 12 is lined with multiple membrane strips 14. Each strip is 0.5mm wide and micrometer-high. It remains 0.5mm thick and maintains its micrometer-high height. Even after immersion in culture medium for five days, it remains intact and does not affect cell proliferation and migration. Gridded lines 15 are located on and below the membrane strips 14, facilitating observation and calculation of relative scratch area. The membrane strips 14 are regularly elongated and feature tear-off zones 141 at their ends. The sterile forceps grip is located on the sidewall of the well plate 13, slightly above the bottom plate 14.
[0031] Membrane strip characteristics: The surface of the membrane strip 14 is made of fibrinogen material, and there will be no colloid residue after tearing; after tearing, the cells in the membrane strip 14 can normally proliferate on the top of the bottom plate 14 without affecting the physiological function of the cells; there is a gap a between the membrane strips 14, such as Figure 2 As shown, the cell expansion is not affected and in the process of tearing the membrane strip 14, the cells on other membrane strips 14 will not be torn off.
[0032] The above embodiments are only for illustrating the technical idea of the present invention and cannot be used to limit the protection scope of the present invention. Any changes made on the basis of the technical solution in accordance with the technical idea proposed by the present invention shall fall within the protection scope of the present invention.
Claims
1. A culture device for cell migration experiment, characterized in that , comprising a bottom plate (11), a well plate (12) on the top of the bottom plate (11), a plurality of culture well positions (13) distributed in a rectangular array on the top of the well plate (12), at least one membrane strip (14) provided at the bottom of the culture well position (13), grid scale lines (15) provided at the top of the membrane strip (14), and grid scale lines (15) also provided at the bottom of the culture well position (13) and at the bottom of the membrane strip (14).
2. A culture device for cell migration experiment according to claim 1, characterized in that: The two ends of the film strip (14) are supported to form tearing areas (141).
3. A culture device for cell migration experiment according to claim 1, characterized in that: Two adjacent membrane strips (14) in the culture well (13) are arranged in parallel, and a gap is provided between the adjacent membrane strips (14).
4. A culture device for cell migration experiment according to claim 1, characterized in that: The surface of the membrane strip (14) is made of fibrinogen.