A method for preparing a temperature-sensitive cell culture device

Abstract

The present application relates to the technical field of cell culture, and discloses a preparation method of a temperature-sensitive cell culture device, which relates to a mold and a culture sheet. One side of the culture sheet is coated with a temperature-sensitive liquid layer. The mold is provided with a mold cavity, and the mold cavity is provided with a groove for accommodating the culture sheet. The preparation method comprises the following steps: placing the culture sheet in the groove, with the side coated with the temperature-sensitive liquid layer facing the bottom of the groove; injecting a molten material into the mold cavity, and the molten material forms the cell culture device after cooling; and when the cell culture device is formed, the culture sheet is located at the bottom of the concave cavity of the cell culture device and is combined with the cell culture device to form an integrated structure. The temperature-sensitive cell culture device is prepared by this method, and no gap is generated between the culture sheet and the cell culture device, the combination effect is good, and the cell culture device and the culture sheet do not need to be subjected to adhesive or hot-pressing treatment, so that the combination efficiency is high.

Description

Technical Field

[0001] This invention relates to the field of cell culture technology, and in particular to a method for preparing a cell culture device. Background Technology

[0002] A thermosensitive cell culture device is used in the field of cell culture. After cell culture is completed, cells and extracellular matrix are harvested by a thermosensitive surface cooling desorption method. The harvesting effect is good and damage to cells and extracellular matrix can be avoided. The structure of the thermosensitive cell culture device can be referred to as a thermosensitive cell culture device disclosed in invention patent CN113025493A, published on June 25, 2021. It includes a culture device body with an upward-facing receiving cavity; a culture plate, which is detachably disposed in the receiving cavity, and a thermosensitive liquid layer is coated on the upper surface of the culture plate.

[0003] According to the instructions for the above scheme, the preparation process of the temperature-sensitive cell culture device is as follows: "The culture strip is evenly coated using a coating machine. After coating, the culture strip is cut according to the specifications of different culture device bodies, and then the culture strip is placed into the receiving cavity to complete the installation." Among them, "the culture strip can be placed into the installation position by means of snap-fit, adhesive, ultrasonic welding or hot pressing."

[0004] In the above-mentioned scheme, the culture slide is placed into the cell culture device by snap-fitting. However, there may be gaps between the culture slide and the cell culture device, resulting in poor combination effect. Culture medium and cells may flow into the gaps, affecting the culture effect. On the other hand, fixing the culture slide into the cell culture device by means of adhesive, ultrasonic welding or hot pressing requires adhesive or hot pressing treatment of the cell culture device and the culture slide, which is cumbersome and has low combination efficiency. Summary of the Invention

[0005] The purpose of this invention is to improve the combined effect and efficiency of culture plates and cell culture devices.

[0006] To achieve the above objectives, the present invention provides a method for preparing a temperature-sensitive cell culture device, involving a mold and a culture plate; wherein, one side of the culture plate is coated with a temperature-sensitive liquid layer; the mold has a cavity for manufacturing a cell culture device with a concave cavity, and the cavity has a groove for accommodating the culture plate; the preparation method includes the following steps:

[0007] The culture plate is placed in the groove, with the side coated with the temperature-sensitive liquid layer facing the bottom of the groove;

[0008] Molten material is injected into the mold cavity, and the molten material cools to form the cell culture device; after the cell culture device is formed, the culture plate is located at the bottom of the cavity of the cell culture device and is combined with the cell culture device to form an integral structure.

[0009] Furthermore, the molten material is PS plastic; the culture sheet is made of PET plastic; and the temperature of the molten material injected into the mold cavity is 200℃~255℃.

[0010] Furthermore, the preparation process of the culture plate includes the following steps:

[0011] Prepare the temperature-sensitive solution;

[0012] The temperature-sensitive liquid is coated onto the surface of the substrate to obtain a temperature-sensitive substrate;

[0013] The temperature-sensitive substrate is cut to form the culture sheet coated with a temperature-sensitive liquid layer.

[0014] Furthermore, the preparation process of the thermosensitive solution is as follows: add the thermosensitive compound and free radical initiator to the solvent, stir to dissolve, and react at 50-150℃ for 5-120 min.

[0015] Furthermore, the coating method for the temperature-sensitive liquid is one of roller coating, atomization, or spraying.

[0016] Furthermore, the mold includes a body, the body includes a front template and a rear template, the front template is provided with a cavity and a flow channel communicating with the cavity; the rear template is provided with a core, and the groove is provided on the side of the core near the front template; after the front template and the rear template are closed, the cavity and the core cooperate to form the mold cavity, and the flow channel is used to inject molten material into the mold cavity.

[0017] Furthermore, the core includes a first core connected to the rear template and a second core sleeved on the outer wall of the first core. The groove is disposed on the first core, and a first annular groove is formed between the first core and the second core. After the front template and the rear template are closed, the first core cooperates with the cavity to form the mold cavity. The front template fits against the second core, and the first annular groove is connected to the mold cavity to form the sidewall of the cell culture device.

[0018] Furthermore, the front mold plate is connected to a front mold core, the cavity is disposed on the front mold core, and the flow channel passes through the front mold core and communicates with the cavity.

[0019] Furthermore, the front mold core is also provided with a second annular groove arranged around the cavity, the inner side of the second annular groove being an open side extending to the cavity, so that the outer wall of the formed cell culture device forms a convex ring; the second annular groove corresponds to the second core.

[0020] Furthermore, the mold also includes a demolding module, which includes a pusher and a drive mechanism. The drive mechanism is used to drive the pusher to move the second core toward the front template so that the formed cell culture device can be demolded.

[0021] The method for preparing a temperature-sensitive cell culture device according to an embodiment of the present invention has the following advantages compared with the prior art:

[0022] In this invention, the mold cavity is provided with a groove. When preparing the cell culture device, the culture plate coated with a temperature-sensitive liquid layer is placed in the groove, and then molten material is injected into the mold cavity. The molten material adheres to the culture plate when forming the cell culture device, so that the cell culture device and the culture plate are combined into an integral structure after molding. By preparing a temperature-sensitive cell culture device in this way, no gaps are generated between the culture plate and the cell culture device, resulting in a good combination effect. In practical applications, since there are no gaps, the culture medium and cells will not flow into the gaps, ensuring the cell culture effect. At the same time, since the cell culture device and the culture plate are combined during the preparation of the cell culture device, compared with the prior art, there is no need to perform adhesive or hot pressing treatment on the cell culture device and the culture plate, resulting in high combination efficiency. Attached Figure Description

[0023] Figure 1 This is a structural diagram of the temperature-sensitive cell culture device according to an embodiment of the present invention;

[0024] Figure 2 This is a structural diagram of the front and rear templates of the mold according to an embodiment of the present invention.

[0025] Figure 3 This is an embodiment of the present invention. Figure 2 Cross-sectional structural diagram;

[0026] Figure 4 This is an embodiment of the present invention. Figure 3 A magnified diagram of G in the middle;

[0027] Figure 5 This is an exploded view of the core and culture plate assembly according to an embodiment of the present invention.

[0028] In the diagram, 100 is the mold; 200 is the culture plate; 300 is the cell culture device; 1 is the front template; 2 is the rear template; 21 is the cooling channel; 3 is the core; 31 is the first core; 32 is the second core; 4 is the front mold core; 5 is the pusher; A is the groove; B is the cavity; C is the runner; D is the first annular groove; E is the second annular groove; F is the water tank; a is the injection mechanism. Detailed Implementation

[0029] The specific embodiments of the present invention will be described in further detail below with reference to the accompanying drawings and examples. The following examples are for illustrative purposes only and are not intended to limit the scope of the invention.

[0030] like Figures 1 to 5 As shown, a preferred embodiment of the present invention discloses a method for preparing a temperature-sensitive cell culture device, involving a mold 100 and a culture plate 200; wherein, one side of the culture plate 200 is coated with a temperature-sensitive liquid layer; the mold 100 has a cavity for manufacturing a cell culture device 300 with a concave cavity, and the cavity has a groove A for accommodating the culture plate 200; the preparation method includes the following steps:

[0031] The culture plate 200 is placed in the groove A, with the side coated with the temperature-sensitive liquid layer facing the bottom of the groove A;

[0032] Molten material is injected into the mold cavity, and the molten material cools to form the cell culture device 300. After the cell culture device 300 is formed, the culture plate 200 is located at the bottom of the cavity of the cell culture device 300 and is integrated with the cell culture device 300 to form an integral structure. The integral cell culture device 300 and culture plate 200 are the temperature-sensitive cell culture device described in this embodiment, and its structure is as follows. Figure 1 As shown;

[0033] Specifically, the molten material adheres to the culture plate 200 during the formation of the cell culture device 300, thereby forming an integral structure with the culture plate 200 after molding. This method of preparing a temperature-sensitive cell culture device eliminates gaps between the culture plate 200 and the cell culture device 300, resulting in a good combination effect. In practical applications, the absence of gaps prevents the culture medium and cells from flowing into any gaps, ensuring effective cell culture. Furthermore, since the cell culture device 300 and culture plate 200 are combined during the fabrication of the cell culture device 300, compared to existing technologies, there is no need for adhesive or hot-pressing treatment of the cell culture device 300 and culture plate 200, resulting in high combination efficiency.

[0034] As a specific implementation of this embodiment, the molten material is injected into the cavity B under the action of the injection mechanism a of the injection molding machine. The injection mechanism a is a needle valve type hot runner. The molten material is PS plastic; the culture sheet 200 is made of PET plastic; and the temperature of the molten material injected into the mold cavity is 200℃~255℃.

[0035] The PS plastic is a polystyrene-based plastic, which refers to a class of plastics whose macromolecular chains include styrene groups, including styrene and its copolymers. Specific varieties include ordinary polystyrene, high-impact polystyrene, expandable polystyrene, and metallocene polystyrene, etc.

[0036] PET plastic material is polyethylene terephthalate, commonly known as polyester resin, which is the most important type of thermoplastic polyester.

[0037] In practical applications, the temperature of molten PS plastic is higher than 200℃ but lower than 255℃. At this temperature, the side of the culture sheet 200 that is in contact with the molten PS plastic is in a slightly molten state. Under the action of injection pressure, the culture sheet 200 in the slightly molten state can better adhere to the molten material, which is conducive to the integration of the culture sheet 200 and the cell culture device 300 to form an integrated structure.

[0038] In this embodiment, the preparation process of the culture plate 200 includes the following steps:

[0039] Step 1: Prepare the temperature-sensitive solution;

[0040] The preparation process of the thermosensitive solution is as follows: add the thermosensitive compound and free radical initiator to the solvent, stir to dissolve, and react at 50-150℃ for 5-120 min; the preparation method of the thermosensitive solution can be referred to the invention patent publication number CN105219644A, which describes a temperature-sensitive cell culture surface and its preparation method.

[0041] Step 2: Apply the temperature-sensitive liquid to the surface of the substrate to obtain a temperature-sensitive substrate; the coating method can be one of roller coating, atomization, or spraying, and this embodiment does not limit this method; taking roller coating as an example, the specific process is coating-drying-washing to remove harmful substances from the surface-drying.

[0042] Step 3: Cut the temperature-sensitive substrate to form the culture plate 200 coated with a temperature-sensitive liquid layer; the shape of the cut depends on the shape of the bottom of the cavity of the cell culture device 300 actually produced, and this embodiment does not limit this; taking the bottom of the cavity of the cell culture device 300 as a circle as an example, the shape of the culture plate 200 is a circle that matches it; taking the bottom of the cavity of the culture device as a hexagon as an example, the shape of the culture plate 200 is a hexagon that matches it.

[0043] In this embodiment, the mold 100 includes a body, which includes a front template 1 and a rear template 2. The front template 1 is provided with a cavity B and a flow channel C communicating with the cavity B. The rear template 2 is provided with a core 3, and a groove A is provided on the side of the core 3 facing the front template 1. After the front template 1 and the rear template 2 are closed, the cavity B and the core 3 cooperate to form the mold cavity, and the flow channel C is used to inject molten material into the mold cavity.

[0044] Furthermore, the core 3 includes a first core 31 connected to the rear template 2 and a second core 32 sleeved on the outer wall of the first core 31. The groove A is disposed on the first core 31, and a first annular groove D is formed between the first core 31 and the second core 32. After the front template 1 and the rear template 2 are closed, the first core 31 cooperates with the cavity B to form the mold cavity. The front template 1 is attached to the second core 32, and the first annular groove D is connected to the mold cavity to form the sidewall of the cell culture device 300.

[0045] The first annular groove D can reduce the depth of cavity B while ensuring that the cavity of cell culture device 300 has a certain depth, making the structure of mold 100 compact and reducing the manufacturing cost of mold 100.

[0046] As a specific implementation of this embodiment, the second core 32 is provided with a first through hole and a second through hole arranged in a stepped manner, wherein the first through hole and the second through hole are arranged concentrically and the area of ​​the first through hole is larger than the area of ​​the second through hole, and a step is formed between the first through hole and the second through hole; in actual use, the first core 31 passes through the second through hole and the first through hole in sequence so that the first core 31 and the second core 32 are combined to form the core 3, wherein the second through hole is fitted with the outer wall of the first core 31, and the inner wall of the first through hole cooperates with the outer wall of the first core 31 to form the first annular groove D.

[0047] The front template 1 is connected to the front mold core 4, the cavity B is disposed on the front mold core 4, and the flow channel C passes through the front mold core 4 and communicates with the cavity B. Specifically, the core 3 is the rear mold core that cooperates with the front mold core 4. In this embodiment, the front mold core 4 is detachably connected to the front template 1, and the rear mold core is detachably connected to the rear template 2, so that the mold 100 can produce temperature-sensitive cell culture devices of different specifications by replacing the front mold core 4 and the rear mold core, thereby improving the versatility of the mold 100 and reducing production costs.

[0048] When the mold 100 is closed, the front mold core 4 fits into the second core 32;

[0049] The mold 100 also includes a demolding module, which includes a pusher 5 and a drive mechanism (not shown in the figure). The drive mechanism is used to drive the pusher 5 to work, so that the second core 32 moves towards the front template 1 to demold the formed cell culture device 300. Specifically, the second core 32 pushes the formed cell culture device 300 to demold through the steps formed between the first through hole and the second through hole.

[0050] The front mold core 4 is also provided with a second annular groove E arranged around the cavity B. The inner side of the second annular groove E is an open side extending to the cavity B, so that the outer wall of the formed cell culture device 300 forms a convex ring; the second annular groove E corresponds to the second core 32.

[0051] The second annular groove E causes the outer wall of the molded cell culture device 300 to form a convex ring structure. In actual use, the convex ring structure makes it easy for the experimenter to hold and move the temperature-sensitive cell culture device. When the cell culture device 300 is demolded, the second core 32 will also push the convex ring structure. That is, the second annular groove E causes the cell culture device 300 to form a convex ring structure, which also facilitates the demolding process.

[0052] The end of the first core 31 that extends into the cavity B is the first end, and the shape of the first end determines the shape of the cavity of the cell culture device 300 to be produced.

[0053] In this embodiment, the first end is a frustum-shaped structure, which allows the cell culture device 300 to have a demolding slope, facilitating demolding. The frustum-shaped structure at one end of the first core 31 is just one specific implementation of this embodiment. This embodiment does not limit the shape of the first end of the first core 31, but determines it according to the cavity shape of the cell culture device 300 to be produced. The shape of the first through hole of the second core 32 is adapted to the shape of the first core 31, so that the inner cavity shape and outer wall shape of the cell culture device 300 are the same.

[0054] In another embodiment, the end of the first core 31 that extends into the cavity B can also be a frustum pyramidal structure, which can also enable the cell culture device 300 to have a demolding slope.

[0055] In this embodiment, the pusher 5 is a pusher plate, which is disposed on the rear template 2 and located between the front template 1 and the rear template 2; the first core 31 passes through the pusher plate and the second core 32 is mounted on the pusher plate; the pusher plate pushes the second core 32 to demold the thermosensitive cell culture device formed.

[0056] The front mold core 4 is also provided with an exhaust groove arranged around the second annular groove E, which is used to allow gas in the mold cavity to be discharged. When the gas is discharged, it is discharged from the gap between the front mold core 4 and the second core 32 into the exhaust groove. The exhaust groove can ensure the molding quality of the cell culture device 300.

[0057] The rear template 2 is also provided with a cooling water channel 21, and the first core 31 is also provided with a water tank F that is connected to the cooling water channel 21. Cooling water enters the water tank F through the cooling water channel 21 to cool the first core 31, thereby realizing the cooling and molding of the molten material.

[0058] The mold 100 also includes an upper fixing plate, a lower fixing plate, and other structures, wherein the front template 1 is connected to the upper fixing plate, and the rear template 2 is connected to the lower fixing plate; the mold 100 described in this embodiment also includes the general structure of molds in the prior art, which will not be described in detail in this embodiment.

[0059] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and substitutions can be made without departing from the technical principles of the present invention, and these improvements and substitutions should also be considered within the scope of protection of the present invention.

Claims

1. A method for preparing a temperature-sensitive cell culture device, comprising a mold and a culture plate; wherein, One side of the culture plate is coated with a temperature-sensitive liquid layer; the mold has a cavity for manufacturing a cell culture device with a concave cavity, characterized in that the cavity has a groove for accommodating the culture plate; the preparation method includes the following steps: The culture plate is placed in the groove, with the side coated with the temperature-sensitive liquid layer facing the bottom of the groove; Molten material is injected into the mold cavity, and the molten material cools to form the cell culture device; after the cell culture device is formed, the culture plate is located at the bottom of the cavity of the cell culture device and is combined with the cell culture device to form an integral structure.

2. The method for preparing the thermosensitive cell culture device according to claim 1, characterized in that, The molten material is PS plastic; the culture sheet is made of PET plastic; the temperature of the molten material injected into the mold cavity is 200℃~255℃.

3. The method for preparing the thermosensitive cell culture device according to claim 1, characterized in that, The preparation process of the culture plate includes the following steps: Prepare the temperature-sensitive solution; The temperature-sensitive liquid is coated onto the surface of the substrate to obtain a temperature-sensitive substrate; The temperature-sensitive substrate is cut to form the culture sheet coated with a temperature-sensitive liquid layer.

4. The method for preparing the thermosensitive cell culture device according to claim 3, characterized in that, The preparation process of the thermosensitive solution is as follows: add the thermosensitive compound and free radical initiator to the solvent, stir to dissolve, and react at 50-150℃ for 5-120 min.

5. The method for preparing the thermosensitive cell culture device according to claim 3, characterized in that, The temperature-sensitive liquid can be applied by one of the following methods: roller coating, atomization, or spraying.

6. The method for preparing the thermosensitive cell culture device according to claim 1, characterized in that, The mold includes a body, which includes a front template and a rear template. The front template has a cavity and a flow channel communicating with the cavity. The rear template has a core, and the groove is located on the side of the core near the front template. After the front template and the rear template are closed, the cavity and the core cooperate to form the mold cavity, and the flow channel is used to inject molten material into the mold cavity.

7. The method for preparing the thermosensitive cell culture device according to claim 6, characterized in that, The core includes a first core connected to the rear template and a second core sleeved on the outer wall of the first core. The groove is disposed on the first core, and a first annular groove is formed between the first core and the second core. After the front template and the rear template are closed, the first core and the cavity cooperate to form the mold cavity. The front template and the second core are attached to each other. The first annular groove is connected to the mold cavity to form the sidewall of the cell culture device.

8. The method for preparing the thermosensitive cell culture device according to claim 7, characterized in that, The front mold plate is connected to the front mold core, the cavity is disposed on the front mold core, and the flow channel passes through the front mold core and communicates with the cavity.

9. The method for preparing the thermosensitive cell culture device according to claim 8, characterized in that, The front mold core is also provided with a second annular groove arranged around the cavity. The inner side of the second annular groove is an open side extending to the cavity, so that the outer wall of the formed cell culture device forms a convex ring. The second annular groove corresponds to the second core.

10. The method for preparing the thermosensitive cell culture device according to claim 9, characterized in that, The mold also includes a demolding module, which includes a pusher and a drive mechanism. The drive mechanism is used to drive the pusher to move the second core toward the front template so that the formed cell culture device can be demolded.

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