A cell culture device
By designing spacer pores and culture tanks in the cell culture device, the problem of uneven cell distribution in traditional cell culture was solved, and stable expansion and efficient culture of monoclonal cells were achieved.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- ZHEJIANG TIANYUAN BIOTECHNOLOGY CO LTD
- Filing Date
- 2025-06-30
- Publication Date
- 2026-05-29
AI Technical Summary
In traditional cell culture, cells are prone to uneven distribution during migration, leading to cell overlap and paradifferentiation, which affects the pluripotency of pluripotent stem cells.
Design a cell culture device including a culture plate and a cover plate. The culture plate has multiple spaced wells, and culture components are placed in the wells. Multiple culture tanks are arranged on the culture components. After cell suspension is inoculated, single cells or small clusters fall into the culture tanks. The structure of the culture tanks restricts the fusion of adjacent cells. The cover plate has a frame to form independent spaces.
It improves the stability and yield of monoclonal cell culture, reduces the probability of cell fusion, and ensures the independence and expansion effect of monoclonal colonies.
Smart Images
Figure CN224299256U_ABST
Abstract
Description
Technical Field
[0001] This utility model relates to the field of medical device technology, and more specifically, to a cell culture device. Background Technology
[0002] Monoclonal expansion of pluripotent stem cells (PSCs) is of great significance in cell therapy and regenerative medicine. In traditional cell culture, six-well plates are typically used as culture containers. Specifically, a cell suspension containing a certain concentration of cell clusters is added to any well of the six-well plate. After the cell suspension is seeded into the well, the cell clusters disperse to the bottom of the well. In practical culture procedures, the six-well plate containing the cell suspension needs to be moved to an incubator for cell culture.
[0003] However, it is difficult to maintain a stable state during migration, which causes cell clusters distributed in the same pore to move and merge with each other, leading to cell overlap and uneven distribution, which indirectly promotes paradifferentiation and affects the pluripotency of stem cells. Utility Model Content
[0004] The technical problem solved by this invention is that after cell seeding, it is difficult to maintain a stable state of movement during the process, which causes cell clusters distributed in the same well to move and easily merge with each other, resulting in cell overlap and uneven distribution, which indirectly promotes paradifferentiation and affects the pluripotency of stem cells.
[0005] To address the aforementioned problems, this utility model provides a cell culture device, which includes a culture plate and a cover plate. The culture plate has a plurality of slots spaced apart from each other. At least one of the slots is provided with a culture component, which includes a plurality of culture tanks spaced apart from each other. The culture tanks are used to contain pre-cultured cells. The cover plate is detachably placed on the culture plate and has a plurality of frames that correspond to the plurality of slots.
[0006] Compared with existing technologies, the technical effects achieved by this solution are as follows: The cell culture device includes a culture plate and a cover plate; the culture plate has multiple wells spaced apart from each other; it is used to contain cell suspension and culture medium, and at least one of the wells is equipped with a culture component, which includes multiple culture tanks spaced apart from each other; in actual operation, the cells to be cultured are dissociated into single cells or small clusters after single-cell treatment, and after a cell suspension with a certain concentration of single cells or small clusters is added to the wells, the single cells or small clusters scatter into the multiple culture tanks on the culture component; by setting the culture component in the wells, and the culture component having multiple culture tanks spaced apart from each other, the traditional cell culture process is overcome by having multiple single cells or small clusters directly distributed on the flat surface at the bottom of the culture plate. On the surface, this design addresses the defect that makes single cells or small clusters prone to cell drift and cell fusion during the early expansion stage. Specifically, when a cell suspension containing multiple single cells or small clusters is seeded into the wells, the single cells or small clusters can fall into the culture tanks on the culture assembly. Due to the structural limitations of the culture tanks, cells in adjacent culture tanks are less likely to fuse, and can expand within the area defined by their respective culture tanks to form independent monoclonal colonies, reducing the probability of cell fusion and improving the stability and yield of monoclonal cell culture. The cell culture device also includes a cover plate, which is detachably placed on the culture plate. The cover plate has multiple frames that correspond to multiple wells, so that each well constitutes an independent space, preventing interference between adjacent wells during the culture process.
[0007] In one embodiment of this utility model, the culture tank is a curved groove.
[0008] Compared with existing technologies, the technical effects achieved by this technical solution are as follows: the culture tank is a curved groove, which allows single cells or small clusters to fall into the culture tank during cell inoculation. Under the action of gravity and liquid surface tension, the single cells or small clusters will preferentially roll to the center of the bottom of the culture tank, so that the single cells or small clusters can start amplification at the center of the bottom of the culture tank and form monoclonal colonies.
[0009] In one embodiment of this utility model, the diameter of the culture tank is 3mm to 4mm; the center height of the culture tank is 0.1mm to 0.2mm.
[0010] Compared with existing technologies, the technical effects achieved by this technology are as follows: the diameter of the culture tank is 3mm~4mm; the center height of the culture tank is 0.1mm~0.2mm, which allows single cells or small clusters to have a certain amplification space in the culture tank. Due to the limited height of the culture tank, single cells or small clusters grow in a two-dimensional monolayer form during cell amplification, and are not easy to stack or form spherical colonies.
[0011] In one embodiment of this utility model, the distance between two adjacent culture tanks in a plurality of culture tanks is greater than 5 mm.
[0012] Compared with existing technologies, the technical effects achieved by this solution are as follows: the distance between two adjacent culture tanks in multiple culture tanks is greater than 5 mm. In conjunction with the actual situation of cell culture, in the process of induced pluripotent stem cell culture, single cells or small clusters can reach passage density after 3 to 6 days of culture. The distance between adjacent culture tanks is greater than 5 mm, which can avoid the fusion of the edges of adjacent monoclonal colonies after they expand outside the culture tank range, thus ensuring the independence of the clones.
[0013] In one embodiment of this invention, the culture component is a cell-compatible material, which is any one of polystyrene, polycarbonate, glass, and polydimethylsiloxane.
[0014] Compared with existing technologies, the technical effects achieved by adopting this solution are as follows: the culture components are made of cell-compatible materials, which helps to improve the efficiency and quality of cell culture. Specifically, polystyrene (PS) has high transparency, which facilitates the observation of cell growth; polycarbonate (PC) has good transparency, which facilitates the observation of cell growth; glass has high transparency and excellent optical properties, which is suitable for high-resolution imaging; and polydimethylsiloxane (PDMS) has low toxicity, allows gas exchange, and is suitable for long-term live cell culture.
[0015] In one embodiment of this invention, the surface of the culture component is provided with a coating layer, which includes an extracellular matrix.
[0016] Compared with existing technologies, the technical effects achieved by this solution are as follows: Pre-coating the surface of the culture component with an extracellular matrix can enhance cell adhesion and proliferation. In some embodiments, the extracellular matrix is any one of Matrigel, Lamin-521, or Vitronectin-XF. Specifically, Matrigel is derived from the basement membrane extract of mouse sarcoma and contains laminin, collagen IV, nestin, growth factors, etc., which can promote cell adhesion and polarization, mimicking the natural matrix. The basal membrane structure supports the polar growth of epithelial cells, endothelial cells, and tumor cells, and can also maintain the pluripotency of stem cells, provide a biomimetic microenvironment for induced pluripotent stem cells and embryonic stem cells, and inhibit spontaneous differentiation, etc.; Lamin-521 can bind to a variety of receptors on the cell surface, including integrins and polysaccharides, which can improve cell survival rate and cloning efficiency; Vitronectin-XF is composed of recombinant human vitronectin and does not contain animal components (Xeno-Free), and mediates cell adhesion through integrin αvβ3 / β5.
[0017] In one embodiment of this invention, the culture component is processed onto the bottom of the culture dish by injection molding or compression molding.
[0018] Compared with existing technologies, the technical effects achieved by adopting this technical solution are as follows: it can achieve the technical effects corresponding to any of the above examples, which will not be elaborated here.
[0019] By adopting the technical solution of the present invention, the following technical effects can be achieved:
[0020] (1) The cell culture device includes a culture plate and a cover plate; the culture plate is provided with multiple wells arranged at intervals; used to contain cell suspension and culture medium, at least one of the multiple wells is provided with a culture component, the culture component includes multiple culture tanks arranged at intervals; in actual operation, the cells to be cultured are dissociated into single cells or small clusters after single-cell treatment, and after adding a cell suspension with a certain concentration of single cells or small clusters to the wells, the single cells or small clusters are scattered in the multiple culture tanks on the culture component; by setting the culture component in the wells, the culture component is provided with multiple culture tanks arranged at intervals, which overcomes the problem that in traditional cell culture, multiple single cells or small clusters are directly distributed on the plane where the bottom of the culture plate is located, so that single cells or small clusters are scattered in the multiple culture tanks on the culture component; In the early expansion stage, cell clusters are prone to cell drift, leading to cell fusion defects. Specifically, when a cell suspension containing multiple single cells or small clusters is seeded into the wells, the single cells or small clusters can fall into the culture tanks on the culture assembly. Due to the structural limitations of the culture tanks, cells in adjacent culture tanks are less likely to fuse, and can expand and form independent monoclonal colonies within the area defined by their respective culture tanks, reducing the probability of cell fusion and improving the stability and yield of monoclonal cell culture. The cell culture device also includes a cover plate, which is detachably placed on the culture plate. The cover plate has multiple frames, which correspond to multiple wells, so that each well constitutes an independent space, preventing interference between adjacent wells during the culture process.
[0021] (2) The culture components of this utility model are applicable to a variety of culture systems, including but not limited to manual culture, automated single-cell screening platform, high-throughput monoclonal screening equipment, etc. Attached Figure Description
[0022] To more clearly illustrate the technical solutions of the embodiments of this utility model, the drawings to be used in the description of the embodiments will be briefly introduced below. Obviously, 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.
[0023] Figure 1 A schematic diagram of the culture plate structure provided in an embodiment of this utility model;
[0024] Figure 2 A schematic diagram of the cover plate structure provided in an embodiment of this utility model;
[0025] Figure 3 This is a first-view structural diagram of the culture component provided in an embodiment of the present invention;
[0026] Figure 4 This is a schematic cross-sectional view of the culture plate provided in an embodiment of the present invention;
[0027] Figure 5 for Figure 4 Enlarged view of a portion of point A in the middle.
[0028] Explanation of reference numerals in the attached figures:
[0029] 100, culture plate; 110, well; 120, culture assembly; 121, culture tank; 200, cover plate; 210, frame. Detailed Implementation
[0030] To make the above-mentioned objectives, features and advantages of this utility model more apparent and understandable, the specific embodiments of this utility model will be described in detail below with reference to the accompanying drawings.
[0031] See Figure 1 , Figure 2 and Figure 3 To address the aforementioned problems, this utility model provides a cell culture device, which includes a culture plate 100 and a cover plate 200. The culture plate 100 is provided with a plurality of slots 110 spaced apart from each other. At least one of the slots 110 is provided with a culture component 120, which includes a plurality of culture tanks 121 spaced apart from each other. The culture tanks 121 are used to contain pre-cultured cells. The cover plate 200 is detachably placed on the culture plate 100, and the cover plate 200 is provided with a plurality of frames 210, which correspond to the plurality of slots 110.
[0032] Further: The cell culture apparatus includes a culture plate 100 and a cover plate 200; the culture plate 100 has a plurality of wells 110 spaced apart from each other; at least one of the wells 110 is provided with a culture component 120, the culture component 120 including a plurality of culture tanks 121 spaced apart from each other; the culture tanks 121 are used to contain pre-cultured cells; by setting the culture component 120 in the wells 110, and the culture component 120 having a plurality of culture tanks 121 spaced apart from each other, the defect of traditional cell culture in which multiple single cells or small clusters are directly distributed on the plane at the bottom of the culture plate 100, making it easy for single cells or small clusters to drift and lead to cell fusion in the early expansion stage, is overcome. Specifically, the culture plate 100 contains multiple single cells or small clusters of cells. When the cell suspension of the cluster is inoculated into the well 110, single cells or small clusters can fall into the culture tank 121 on the culture component 120. Due to the structural limitation of the culture tank 121, cells in adjacent culture tanks 121 are less likely to fuse and can expand and form independent monoclonal colonies within the area defined by their respective culture tanks 121, reducing the probability of cell fusion and improving the stability and yield of monoclonal cell culture. The cell culture device also includes a cover plate 200, which is detachably placed on the culture plate 100. The cover plate 200 is provided with multiple frames 210, which correspond to multiple wells 110, so that each well 110 constitutes an independent space and prevents interference between adjacent wells 110 during the culture process.
[0033] See Figure 4 and Figure 5 In a specific embodiment of this utility model, the culture tank 121 is a curved groove.
[0034] Furthermore, in some embodiments, the culture tank 121 is a curved groove; during the cell seeding process, after a single cell or small cluster falls into the culture tank 121, under the action of gravity and liquid surface tension, the single cell or small cluster preferentially rolls to the center position of the bottom of the culture tank 121, so that the single cell or small cluster starts to amplify at the center position of the bottom of the culture tank 121, forming a monoclonal colony.
[0035] In a specific embodiment of this utility model, the diameter of the culture tank 121 is 3mm to 4mm; the center height of the culture tank 121 is 0.1mm to 0.2mm.
[0036] Furthermore, in some embodiments, the diameter of the culture tank 121 is 3.5 mm and the center height of the culture tank 121 is 0.1 mm, so that single cells or small clusters have a certain amplification space in the culture tank 121. Due to the limited height of the culture tank 121, single cells or small clusters grow in a two-dimensional monolayer form during cell amplification, and are not easy to stack or form spherical colonies.
[0037] In a specific embodiment of this utility model, the distance between two adjacent culture tanks 121 in the plurality of culture tanks 121 is greater than 5 mm.
[0038] Furthermore, the number of culture tanks 121 can be adjusted according to the actual culture conditions. In some embodiments, the number of culture tanks 121 is 6 to 10, and the distance between two adjacent culture tanks 121 is greater than 5 mm. In conjunction with the actual situation of cell culture, during the culture of induced pluripotent stem cells, single cells or small clusters can reach the passage density after 3 to 6 days of culture. The distance between adjacent culture tanks 121 is greater than 5 mm, which can prevent the edges of adjacent monoclonal colonies from merging and interfering with each other after the monoclonal colonies expand beyond the range of the culture tank 121, thereby ensuring the independence of the monoclonal colonies.
[0039] In a specific embodiment of this utility model, the culture component 120 is a cell-compatible material, which is any one of polystyrene, polycarbonate, glass and polydimethylsiloxane.
[0040] Furthermore, the culture component 120 is made of cell-compatible materials, which helps to improve the efficiency and quality of cell culture; specifically, polystyrene (PS) has high transparency, which facilitates the observation of cell growth; polycarbonate (PC) has good transparency, which facilitates the observation of cell growth; glass has high transparency and good optical properties, which is suitable for high-resolution imaging; and polydimethylsiloxane (PDMS) has low toxicity, allows gas exchange, and is suitable for long-term live cell culture.
[0041] In a specific embodiment of this utility model, the surface of the culture component 120 is provided with a coating layer, which includes an extracellular matrix.
[0042] Furthermore, pre-coating the surface of the culture component 120 with an extracellular matrix can enhance cell adhesion and proliferation. In some embodiments, the extracellular matrix is any one of Matrigel, Lamin-521, or Vitronectin-XF. Specifically, Matrigel is derived from the basement membrane extract of mouse sarcoma and contains laminin, collagen IV, nestin, growth factors, etc., which can promote cell adhesion and polarization, mimic the structure of the natural basement membrane, and support cell growth. The polar growth of epithelial cells, endothelial cells, and tumor cells can also maintain the pluripotency of stem cells, provide a biomimetic microenvironment for induced pluripotent stem cells and embryonic stem cells, and inhibit spontaneous differentiation; Lamin-521 can bind to a variety of receptors on the cell surface, including integrins and polysaccharides, which can improve cell survival and cloning efficiency; Vitronectin-XF is composed of recombinant human vitronectin and does not contain animal components (Xeno-Free), and mediates cell adhesion through integrin αvβ3 / β5.
[0043] Furthermore, in some embodiments, the cell culture device is a six-well plate, wherein all six culture dishes are equipped with culture components 120.
[0044] In a specific embodiment of this utility model, the culture component 120 is processed onto the bottom of the culture dish by injection molding or compression molding.
[0045] While the present invention has been disclosed above, it is not limited thereto. Any person skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention; therefore, the scope of protection of the present invention should be determined by the scope defined in the claims.
Claims
1. A cell culture device, characterized in that, The cell culture device includes a culture plate (100) and a cover plate (200), wherein the culture plate (100) is provided with a plurality of slots (110) spaced apart from each other; At least one of the plurality of pores (110) is provided with a culture assembly (120), the culture assembly (120) including a plurality of culture tanks (121) arranged at intervals from each other; the culture tanks (121) are used to contain pre-cultured cells; The cover plate (200) is detachably placed on the culture plate (100). The cover plate (200) is provided with a plurality of frames (210), which correspond to the plurality of holes (110).
2. The cell culture apparatus according to claim 1, characterized in that, The culture tank (121) is a curved groove.
3. The cell culture apparatus according to claim 2, characterized in that, The diameter of the culture tank (121) is 3mm~4mm; The center height of the culture tank (121) is 0.1mm~0.2mm.
4. The cell culture apparatus according to claim 1, characterized in that, The distance between two adjacent culture tanks (121) in the plurality of culture tanks (121) is greater than 5 mm.
5. The cell culture apparatus according to claim 1, characterized in that, The culture component (120) is a cell-compatible material; the cell-compatible material is any one of polystyrene, polycarbonate, glass and polydimethylsiloxane.
6. The cell culture apparatus according to claim 1, characterized in that, The culture component (120) has a coating layer on its surface, the coating layer including an extracellular matrix.
7. The cell culture apparatus according to claim 1, characterized in that, The culture component (120) is processed onto the bottom of the cavity (110) by injection molding or compression molding.