Adhesive cell culture container

Abstract

The utility model belongs to the technical field of cell culture devices, and relates to an adherent cell culture container. The adherent cell culture container comprises a bottom plate and side plates formed by extending upwards from the bottom plate, and a containing space for containing a culture solution is defined by the bottom plate and the side plates; a plurality of cultivation grooves for accommodating adherent cells are formed in the bottom plate, and each cultivation groove comprises a bottom wall, a vertical part connected with the bottom wall and an opening part which is connected with the vertical part and has a gradually expanded cross section. When the adherent cells are cultured, the adherent cells proliferate on the bottom wall and the vertical part to form a stable foundation, and then the adherent cells enter the opening part. The side wall of the opening part is slower than that of the vertical part, so that the adherent cells can rapidly proliferate along the side wall of the opening part, the contact area with the container is enlarged, and the stable foundation and the larger contact area are beneficial to keeping the cells stable during liquid change.

Description

Technical Field

[0001] The utility model belongs to the technical field of cell culture devices and relates to an adherent cell culture container. Background Art

[0002] Adherent cells are cells that need to attach to a fixed support or the bottom of a container when cultured in vitro. Adherent cells rely on a solid surface for growth and division, which is essential for their normal physiological activities and cell differentiation.

[0003] In general cell culture vessels, operations such as changing the medium often disturb the tissue masses of adherent cells. During the medium change process, the movement and replacement of the culture medium often physically disturb the adherent cells, causing the adherent cells to fall off or be damaged from the bottom wall of the culture vessel, reducing the survival rate.

[0004] In order to reduce this disturbance, one improvement measure is to open specific grooves on the bottom wall of the cell culture container and use these grooves to culture adherent cells. In this way, the contact area between the adherent cells and the bottom wall can be increased to a certain extent, thereby increasing the attachment stability of the adherent cells. However, the stability of this method in actual culture is still insufficient, especially during the proliferation of adherent cells, the increase in space demand caused by the proliferation of adherent cells is not fully considered, thereby limiting the stability and growth efficiency of the adherent cells in the groove. Utility Model Content

[0005] The utility model aims to provide an adherent cell culture container which combines the proliferation of adherent cells to reduce the disturbance of adherent cells during operations such as pipetting.

[0006] To achieve the above object, the utility model provides the following technical solutions: an adherent cell culture container, comprising a bottom plate and side plates extending upward from the bottom plate, wherein the bottom plate and the side plates surround a containing space for containing a culture medium;

[0007] The bottom plate is provided with a plurality of culture grooves for accommodating adherent cells, and each of the culture grooves comprises a bottom wall, a vertical portion connected to the bottom wall, and an open portion connected to the vertical portion and having a gradually expanding cross section.

[0008] Furthermore, the cross-sectional shape of the vertical portion and the open portion on the horizontal plane is circular, elliptical or rectangular.

[0009] Furthermore, the cross-sections of the vertical portion and the open portion in the horizontal plane are both circular, and the cross-sectional diameter ranges from 5 mm to 20 mm.

[0010] Furthermore, the depth of the cultivation tank is any value between 5 mm and 15 mm.

[0011] Furthermore, the height ratio of the vertical portion to the open portion is set at any value between 1:1 and 1:3.

[0012] Furthermore, the spacing between adjacent cultivation grooves is any value between 5 mm and 15 mm.

[0013] Furthermore, the bottom wall is a plane.

[0014] Furthermore, the bottom wall is a curved surface that is concave downward.

[0015] Furthermore, the bottom plate and side plates of the adherent cell culture container are integrally formed.

[0016] The beneficial effects of the utility model are as follows: first, by providing a bottom wall and a vertical portion, the present application enables adherent cells to proliferate thereon, thereby forming a stable foundation; then, the adherent cells continue to proliferate, and after entering the open portion, due to the gradual expansion of the cross-section of the open portion, the slope of the side wall of the open portion becomes gentler than that of the side wall of the vertical portion, thereby enabling the adherent cells to grow rapidly along the side wall of the open portion, and the contact area between the adherent cells and the container increases rapidly; finally, the stable foundation and the larger contact area make the adherent cells less susceptible to disturbance during fluid replacement.

[0017] The above description is only an overview of the technical solution of the utility model. In order to more clearly understand the technical means of the utility model and implement it according to the contents of the specification, the following is a detailed description of the preferred embodiments of the utility model in conjunction with the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a schematic structural diagram of an adherent cell culture container shown in one embodiment of the utility model;

[0019] Figure 2 for Figure 1 Top view of the medium adherent cell culture vessel;

[0020] Figure 3 for Figure 2 A cross-sectional view of a structure along the AA direction;

[0021] Figure 4 for Figure 2 A cross-sectional view of another structure along the AA direction;

[0022] Figure 5 for Figure 2 A cross-sectional view of another structure along the AA direction. DETAILED DESCRIPTION

[0023] The technical solution of the utility model will be described clearly and completely below in conjunction with the accompanying drawings. Obviously, the described embodiments are part of the embodiments of the utility model, not all of the embodiments. Based on the embodiments of the utility model, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the utility model.

[0024] In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc., indicating the orientation or positional relationship, are based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as limiting the present invention. In addition, the terms "first", "second", and "third" are used for descriptive purposes only, and cannot be understood as indicating or implying relative importance.

[0025] In the description of the present invention, it should be noted that, unless otherwise clearly specified and limited, the terms "installation", "connection" and "connection" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0026] In addition, the technical features involved in the different embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0027] See also Figures 1 to 3 The adherent cell culture container 100 shown in a preferred embodiment of the present application comprises a bottom plate 1 and a side plate 2 extending upward from the bottom plate 1, wherein the bottom plate 1 and the side plate 2 surround a containing space 3 for containing a culture medium. The adherent cells are placed on the bottom plate 1 for culture, and all the adherent cells share the culture medium in the containing space 3.

[0028] A plurality of culture tanks 21 for accommodating adherent cells are formed on the bottom plate 2. Each culture tank 21 includes a bottom wall 211, a vertical portion 212 connected to the bottom wall 211, and an open portion 213 connected to the vertical portion 212 and having a gradually expanding cross section.

[0029] Specifically, the vertical portion 212 includes a first side wall 2121 extending upward from the periphery of the bottom wall 211, and a vertical space 2122 formed by the first side wall 2121, and the first side wall 2121 is perpendicular to the bottom wall 211. The open portion 213 includes a second side wall 2131 extending upward and outward from the top end of the first side wall 2121, and an open space 2132 formed by the second side wall 2131.

[0030] The cross-sectional shape of the vertical portion 212 and the open portion 213 on the horizontal plane is circular, elliptical or rectangular.

[0031] In one embodiment, the cross-sections of the vertical portion 212 and the open portion 213 on the horizontal plane are both circular, and the cross-sectional diameter ranges from 5 mm to 20 mm. The vertical portion 212 is a cylinder, and the open portion 213 is a truncated cone, and the cross-sectional diameter of the bottom of the open portion 213 is equal to the cross-sectional diameter of the vertical portion 212. The cross-sectional diameter of the bottom of the open portion 213 is greater than 5 mm, and the cross-sectional diameter of the top is less than 20 mm. See Figure 4 Optionally, the second side wall 2131 of the open portion 213 can extend upward along a curve with a gradually increasing curvature, so that the slope of the portion where the second side wall 2131 is connected to the first side wall 2121 is relatively gentle, and the adherent cells can proliferate rapidly, rapidly increasing the contact area with the adherent cell culture container 100. The circular cross section can evenly distribute the adherent cells and the culture fluid, reduce the accumulation and dead corners of the cells during the culture process, and promote the uniform growth of the cells.

[0032] In another embodiment, the cross-sections of the vertical portion 212 and the open portion 213 on the horizontal plane are both rectangular. Among them, the vertical portion 212 is a cuboid, the open portion 213 is a prism, and the bottom of the open portion 213 coincides with the top of the vertical portion 212. More preferably, the cross-sections of the open portion 213 and the vertical portion 212 on the horizontal plane are square, so as to facilitate the uniform distribution of the adherent cells. Optionally, the second side wall 2131 of the open portion 213 can also extend upward along the curve, and the curvature gradually increases, so that the slope of the portion where the second side wall 2131 is connected to the first side wall 2121 is relatively gentle, and the adherent cells can proliferate rapidly, and the contact area with the adherent cell culture container 100 is rapidly increased. The rectangular cross section provides a large flat surface area, which is conducive to the spreading and attachment of cells, and is convenient for microscopic observation and analysis. As for the numerical range of the length and width of the rectangular cross section, it can be adjusted according to the actual situation based on the circular cross section, which will not be described in detail here.

[0033] In another embodiment, the cross-sections of the vertical portion 212 and the open portion 213 in the horizontal plane are both elliptical. The elliptical cross-section combines the advantages of the circular and rectangular shapes, provides a larger contact area, and maintains good fluid dynamics, which is conducive to the flow of the culture solution and the supply of cell nutrients. The elliptical cross-section scheme is similar to the structure in the first two embodiments and will not be described in detail here.

[0034] In another embodiment, the cross section of the vertical portion 212 in the horizontal plane is rectangular, the cross section of the open portion 213 in the horizontal plane is circular, and the horizontal projection of the vertical portion 212 is located within the horizontal projection of the cross section of the bottom of the open portion 213. More preferably, the cross section of the vertical portion 212 is square, and the top cross section of the vertical portion 212 is inscribed in the bottom cross section of the open portion 213, so as to facilitate the smooth transition of the vertical portion 212 to the open portion 213. This combined cross section can provide a stable attachment foundation through the vertical portion 212 with a rectangular cross section, and at the same time promote the rapid growth and uniform distribution of cells through the gradual expansion characteristics of the open portion with a circular cross section.

[0035] The depth of the culture tank 21 is any value between 5 mm and 15 mm. For some smaller adherent cells, such as epithelial cells, the depth can be set to 5 mm, so that the adherent cells grow quickly and emerge from the culture tank 21. For adherent cells with medium requirements, such as fibroblasts, the depth can be set to 10 mm. The moderate depth balances the nutrient supply and cell growth space, which is suitable for the routine culture of most adherent cells. For adherent cells that require a large number of cells or long-term culture, such as stem cells, the depth can be set to 15 mm, so that the adherent cells can obtain sufficient growth space and time in the culture tank.

[0036] The height ratio of the vertical portion 212 to the open portion 213 is set to any value between 1:1 and 1:3. When the ratio is 1:1, the heights of the vertical portion 212 and the open portion 213 are equal, ensuring that the adherent cells can obtain sufficient growth space in both the vertical portion 212 and the open portion 213, which is conducive to the uniform distribution of the adherent cells. When the ratio is 1:2, the height of the open portion 213 is twice that of the vertical portion 212, providing a larger open space 2132, which is conducive to increasing the contact area between the adherent cells and the adherent cell culture container 100. When the ratio is 1:2, the height of the open portion 213 is three times that of the vertical portion 212, further increasing the contact area between the adherent cells and the adherent cell culture container 100, ensuring that the adherent cells are more stable when changing the liquid.

[0037] The spacing between adjacent culture tanks 21 is any value between 5 mm and 15 mm. Generally, 5 mm, 10 mm and 15 mm can be selected. According to the cross-sectional size of the top of the open portion 213, a suitable spacing size is selected to improve the efficiency of cell culture and the operability of the experiment.

[0038] In one embodiment, the bottom wall 211 is a plane, and the adherent cells are evenly distributed on the bottom wall 211, which is beneficial to the uniformity of the adherent cell growth and the convenience of subsequent operations.

[0039] See also Figure 5 In another example, the bottom wall 211 is a curved surface that is concave downward, and the adherent cells gather toward the lowest end of the bottom wall 211 and proliferate upward from the lowest end, thereby increasing the density of the adherent cells in a unit area.

[0040] The bottom plate 2 and the side plate 1 of the adherent cell culture container 100 are integrally formed by injection molding and are directly formed. In addition, for the convenience of observation, they are generally made of glass or plastic.

[0041] Taking the cultivation of umbilical cord mesenchymal stem cells as an example, the workflow of the adherent cell culture container 100 is described as follows:

[0042] First, obtain the umbilical cord tissue block to be cultivated, place the umbilical cord tissue block on the bottom wall 211, and add culture medium. The umbilical cord mesenchymal stem cells crawl out from the surrounding of the umbilical cord tissue block and proliferate upward along the first side wall 2121 and the second side wall 2131. In addition, in the process of the umbilical cord mesenchymal stem cells proliferating upward, they will also proliferate horizontally, so that they can fill the vertical space 2122, forming a stable foundation, and cooperate with the first side wall 2121, so that shaking is not allowed. In addition, when the umbilical cord mesenchymal stem cells enter the second side wall 2131 from the first side wall 2121, the slope slows down, so that they can quickly expand and proliferate outward, increase the contact area between the umbilical cord mesenchymal stem cells and the adherent cell culture container 100, and further improve stability. Finally, when the umbilical cord mesenchymal stem cells proliferate to the desired size of the cell colony, the next step is performed. During the cultivation process of the adherent cell culture container 100, when the medium is replaced, the umbilical cord mesenchymal stem cells form a stable foundation and a larger contact area with the adherent cell culture container 100, so that the umbilical cord mesenchymal stem cells are not easily disturbed during the medium replacement.

[0043] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0044] The above-mentioned embodiments only express several implementation methods of the utility model, and the description is relatively specific and detailed, but it cannot be understood as limiting the scope of the utility model patent. It should be pointed out that for ordinary technicians in this field, several modifications and improvements can be made without departing from the concept of the utility model, which all belong to the protection scope of the utility model. Therefore, the protection scope of the utility model patent shall be based on the attached claims.

Claims

1. An adherent cell culture container, characterized in that: It comprises a bottom plate and side plates extending upward from the bottom plate, wherein the bottom plate and the side plates surround a containing space for containing culture liquid; The bottom plate is provided with a plurality of culture grooves for accommodating adherent cells, and each of the culture grooves comprises a bottom wall, a vertical portion connected to the bottom wall, and an open portion connected to the vertical portion and having a gradually expanding cross section.

2. The adherent cell culture container according to claim 1, characterized in that: The cross-sectional shape of the vertical portion and the open portion on the horizontal plane is circular, elliptical or rectangular.

3. The adherent cell culture container according to claim 2, characterized in that: The cross-sections of the vertical portion and the open portion on the horizontal plane are both circular, and the cross-sectional diameter ranges from 5 mm to 20 mm.

4. The adherent cell culture container according to claim 3, characterized in that: The spacing between adjacent cultivation grooves is any value between 5 mm and 15 mm.

5. The adherent cell culture container according to claim 4, characterized in that: The depth of the cultivation tank is any value between 5 mm and 15 mm.

6. The adherent cell culture container according to claim 5, characterized in that: The height ratio of the vertical portion to the open portion is set to any value between 1:1 and 1:

3.

7. The adherent cell culture container according to any one of claims 1 to 6, characterized in that: The bottom wall is a plane.

8. The adherent cell culture container according to any one of claims 1 to 6, characterized in that: The bottom wall is a curved surface that is concave downwards.

9. The adherent cell culture container according to any one of claims 1 to 6, characterized in that: The bottom plate and the side plates of the adherent cell culture container are integrally formed.

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