Co-substrate double-layer cell culture device, pore plate and incubator
By inoculating different cells in batches on the culture dish of the cell culture device and using fixed clips to maintain the scaffold morphology, the cytotoxicity, activity damage and scaffold deformation problems that may be caused by cell mixing printing during cell culture, and the stability of double-cell bilayer culture and scaffolds are achieved, making it easier for cell observation.
Patent Information
- Application Number
- CN202323662254.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2023-12-29
- Publication Date
- 2025-03-07
- Estimated Expiration
- 2033-12-29
AI Technical Summary
In the prior art, the cell mixing printing method may lead to problems such as cytotoxicity, impaired cell activity, cell contamination and scaffold deformation during cell culture.
A common basal bilayer cell culture device is provided, including body and fixing clips. By inoculating different cells in batches on the culture dish, the use of traditional cell mixing printing methods is directly avoided. The fixing clips keep the morphology of the scaffold stable and suitable for cell observation.
Double-cell bilayer culture was achieved, avoiding cytotoxicity and activity damage, maintaining the stability of the scaffold, facilitating cell observation, and improving the effect of cell culture.
Smart Images
Figure CN222574669U_ABST
Abstract
Description
Technical Field
[0001] The utility model relates to the technical field of cell culture, and more specifically to a common substrate double-layer cell culture device, a well plate and a culture box. Background Art
[0002] In the process of biological 3D printing simulated skin, the same scaffold is needed as a substrate for multi-layer culture of two types of cells. In the prior art, cells are mainly cultured by cell mixed printing, that is, different cells are mixed in the same biological ink, and the biological ink is replaced during the printing process to achieve layered printing and culture of two types of cells.
[0003] However, the application of this method will cause the following problems: some initiators added to the mixed bio-ink have certain cytotoxicity before the reaction, and may cause damage to cells during the mixing process; after printing, the uncured bio-ink needs to be soaked or rinsed to remove it. During the process, reagents such as alcohol and PBS buffer may be used. The solvents and methods used for removal may cause damage to the cells; during the printing process, the cells need to leave the incubator for a long time, and the printing process does not have the conditions for cell survival, which may affect cell activity; the shear force or curing light during the printing process will affect the cells; during the printing process, the cells are in close contact with the environment, which is easy to cause contamination. Utility Model Content
[0004] In order to solve the problems existing in the above-mentioned background technology, the utility model provides a common substrate double-layer cell culture device, a well plate and a culture box.
[0005] In order to solve the above technical problems, the technical solution adopted by the utility model is:
[0006] In a first aspect, the utility model provides a common substrate double-layer cell culture device, comprising a main body and a fixing clamp, wherein the main body comprises a first shell, a second shell, and a culture dish, wherein the first shell and the second shell are hollow inside, the first surface of the culture dish abuts against one end of the first shell, and the other surface of the culture dish abuts against one end of the second shell; the fixing clamp comprises an annular belt body, and the fixing clamp is sleeved on the outside of the main body through the annular belt body to fix the first shell, the second shell and the culture dish.
[0007] In some embodiments of the present application, the first shell and the second shell are cylindrical.
[0008] In some embodiments of the present application, a surface of the first shell facing the second shell and a surface of the second shell facing the first shell are both provided with protrusions.
[0009] In some embodiments of the present application, the fixing clamp further includes a fixing assembly, and the fixing assembly includes a fixing piece, and the fixing piece is passed through two ends of the annular belt body to fix the fixing clamp.
[0010] In some embodiments of the present application, the fixing assembly also includes a nut, the fixing member is a bolt, the bolt passes through both ends of the annular belt body, the nut is sleeved on the bolt, and the nut is rotated to increase or decrease the diameter of the annular belt body.
[0011] In some embodiments of the present application, the fixing clamp further includes a supporting portion, and the supporting portion is disposed between the annular belt bodies to support the main body.
[0012] In some embodiments of the present application, a hydrophobic coating is provided on the inner walls of the first shell and the second shell.
[0013] In a second aspect, the utility model provides a well plate, including the above-mentioned common substrate double-layer cell culture device, wherein the well plate is provided with a plurality of holes compatible with the common substrate double-layer cell culture device, and the common substrate double-layer cell culture device is arranged in the holes.
[0014] In some embodiments of the present application, a plurality of covers for closing the holes are disposed on the plurality of holes.
[0015] In a third aspect, the present application provides a culture box, comprising a box body and a well plate, wherein the well plate is arranged in the box body.
[0016] Compared with the prior art, the utility model realizes double-cell double-layer culture by inoculating different cells in the culture dish in batches, directly avoiding a series of problems that may be caused by the traditional cell mixing printing method. The fixing clamp can maintain the shape of the bracket, ensure the stability of the bracket during culture, transportation and detection, and avoid the bracket from bending, folding and other deformations due to factors such as immersion in the culture medium, which may affect cell growth; at the same time, this device can better observe the state of the bracket cells, and will not be affected by the position of the microscope lens when observing under the microscope, and different types of cells can be observed by directly turning it upside down. BRIEF DESCRIPTION OF THE DRAWINGS
[0017] Figure 1 Schematic diagram of the structure of the co-substrate double-layer cell culture device in the embodiment of the present application.
[0018] Figure 2 It is an exploded view of the co-substrate double-layer cell culture device in an embodiment of the present application.
[0019] Figure 3 It is a schematic diagram of the structure of the fixing clip of the locking member in the embodiment of the present application.
[0020] Description of reference numerals:
[0021] 1-body; 101-first shell; 102-second shell;
[0022] 2-fixing clip; 201-annular belt body; 202-bolt; 203-nut; 204-supporting part;
[0023] 3-Petri dish. DETAILED DESCRIPTION
[0024] Example 1
[0025] This embodiment provides a common substrate double-layer cell culture device, please refer to Figure 1-3 A common substrate double-layer cell culture device comprises a main body 1 and a fixing clip 2, wherein the main body 1 comprises a first shell 101, a second shell 102, and a culture dish 3, wherein the first shell 101 and the second shell 102 are hollow inside, the first surface of the culture dish 3 abuts against one end of the first shell 101, and the other surface of the culture dish 3 abuts against one end of the second shell 102; the fixing clip 2 comprises an annular belt body 201, and the fixing clip 2 is sleeved on the outer side of the main body 1 through the annular belt body 201 to fix the first shell 101, the second shell 102 and the culture dish 3.
[0026] Specifically, the culture dish 3 is used to place the cell scaffold to be cultured, and the culture dish 3 is arranged between the first shell 101 and the second shell 102. Since the first shell 101 and the second shell 102 are hollow inside, relevant personnel can observe the two sides of the culture dish 3 through the first shell 101 and the second shell 102; the fixing clip 2 includes an annular belt body 201, and the annular belt body 201 is sleeved on the first shell 101, the culture dish 3 and the second shell 102, and the above three parts are arranged in one piece.
[0027] In the specific use process, the first step is to configure the biological ink and use the 3D printer to print the model. After taking it out, use PBS buffer to soak and rinse the bracket and place it in a clean culture dish 3; the second step is to place the culture dish 3 with the bracket under ultraviolet light for 30 minutes; the third step is to place the culture dish 3 between the first shell 101 and the second shell 102, and then fix the first shell 101, the second shell 102 and the culture dish 3 by the annular belt body 201 of the fixing clip 2, and perform double-layer culture on the two sides of the culture dish 3.
[0028] By arranging the culture dish 3 between the first shell 101 and the second shell 102, double-cell double-layer culture can be achieved by inoculating different cells in the culture dish 3 in batches, directly avoiding a series of problems that may be caused by using the traditional cell mixing printing method. For example, the use of mixed ink in the traditional cell mixing and printing method may affect the activity of cells and cause damage to them; the fixing clamp 2 can maintain the shape of the bracket, ensure the stability of the bracket during the culture, transportation and detection process, and avoid the bracket from bending and folding due to factors such as immersion in the culture medium, thereby affecting cell growth; at the same time, the state of the bracket cells can be better observed, and it will not be affected by the position of the microscope lens when observing under the microscope, and different types of cells can be observed by directly turning it upside down.
[0029] Preferably, the first shell 101 and the second shell 102 are cylindrical, and the first shell 101 and the second shell 102 are hollow cylinders. The fixing clamp 2 can better fix the hollow cylinders. At the same time, the cylindrical shape can better adapt to the equipment required for subsequent processes.
[0030] In some embodiments, a protrusion is provided on one side of the first shell 101 facing the second shell 102 and on one side of the second shell 102 facing the first shell 101. The plane area of the first shell 101 and the second shell 102 where the protrusion is provided is larger than the plane area of the other end thereof. The protrusion is provided on the first shell 101 and the second shell 102 to increase the contact area between the two shells, so that the culture dish 3 is better installed on the body 1. At the same time, the protrusion extends outward to increase the contact area while reducing interference with the internal observation field.
[0031] In one embodiment, the fixing clamp 2 further includes a fixing assembly, and the fixing assembly includes a fixing piece, and the fixing piece is passed through the two ends of the annular belt body 201 to fix the fixing clamp 2. The fixing piece fixes the annular belt body 201 of the fixing clamp 2. Further, the fixing assembly also includes a nut 203, and the fixing piece is a bolt 202, and the bolt 202 passes through the two ends of the annular belt body 201. The nut 203 is rotated to increase or decrease the diameter of the annular belt body 201. The annular belt body 201 of the fixing clamp 2 is fixed by the bolt 202, and the nut 203 is rotated to adjust its tightness to better fix the main body 1. It can be understood that the fixing piece is not limited to the bolt 202, and can also be fixed in other ways, as long as the fixing effect is achieved.
[0032] Preferably, the fixing clip 2 further includes a support portion 204, and the support portion 204 is disposed between the annular belt body 201 to support the body 1. The support portion 204 is disposed on one side of the annular belt body 201 opposite to the fixing assembly. The common substrate double-layer cell culture device generally needs to be used in conjunction with a well plate, and the support portion 204 can provide a good supporting effect on the body 1. At the same time, the support portion 204 can also be used to conveniently flip the common substrate double-layer cell culture device for easy observation on a microscope.
[0033] Preferably, a hydrophobic coating is provided on the inner walls of the first shell 101 and the second shell 102, and a special super-hydrophobic coating is coated on the inner walls of the first shell 101 and the second shell 102, which can effectively prevent cells from adhering to the side walls and growing upward during the growth process, ensuring that all cells grow in the middle support position, making subsequent cell-related detection more accurate.
[0034] Example 2
[0035] This embodiment provides a well plate, including a common substrate double-layer cell culture device, wherein the well plate is provided with a plurality of holes adapted to the common substrate double-layer cell culture device, and the common substrate double-layer cell culture device is arranged in the holes. The size of the well plate is adapted to the common substrate double-layer cell culture device, and preferably, the well plate is a twelve-well plate, which can realize double-cell double-layer culture by inoculating different cells in batches on a culture dish 3, directly avoiding a series of problems that may be caused by using a traditional cell mixing printing method, and the fixing clamp 2 can maintain the shape of the support, ensure the stability of the support during the culture, transportation and detection process, and avoid the support from bending, folding and other deformations due to factors such as immersion in the culture medium, which affects cell growth; at the same time, the state of the support cells can be better observed, and it will not be affected by the position of the microscope lens when observing under a microscope, and different types of cells can be observed by directly turning it upside down.
[0036] Preferably, a plurality of covers for closing the holes are provided on the plurality of holes. The covers can close the holes, and after the common substrate double-layer cell culture device is placed in the holes, the covers are closed to better cultivate the cells.
[0037] The specific usage is:
[0038] 1. Prepare the bio-ink and use a 3D printer to print the model. After taking it out, soak and rinse the scaffold with PBS buffer and place it in a clean culture dish 3.
[0039] 2. Place the culture dish 3 with the bracket under ultraviolet light for 30 minutes.
[0040] 3. Use round-tipped tweezers to take out the bracket in the clean bench, place it between the first shell 101 and the second shell 102 as a sandwich, and use the fixing clip 2 to fix it and ensure that the bracket cannot move.
[0041] 4. Place the common substrate double-layer cell culture device into the holes of a twelve-well plate in an ultra-clean bench, drop the cell suspension obtained by cell passage in the ultra-clean bench into the first shell 101 layer of the device, cover the twelve-well plate with a cover, and place it in an incubator.
[0042] 5. After 4 hours of culture, take out the 12-well plate, take out the device in the clean bench, remove the culture medium in the upper layer, add fresh culture medium to the new wells, turn the device upside down and put it into the wells to perform double-layer cell culture.
[0043] Example 3
[0044] The present embodiment provides a culture box, including conventional structures such as a box body and a well plate, wherein a common base double-layer cell culture device is provided in the well plate, and the well plate is arranged in the box body. The well plate and the common base double-layer cell culture device are placed in the culture box for culture, providing a good growth environment for cells, which can make the scaffold culture effect better and accelerate its culture process. This culture box can achieve double-cell double-layer culture by inoculating different cells on the culture dish 3 in batches, directly avoiding a series of problems that may be caused by using the traditional cell mixing printing method. The fixing clip can maintain the shape of the scaffold, ensure the stability of the scaffold during the culture, transportation and detection process, and avoid the scaffold from bending, folding and other deformations due to factors such as immersion in the culture medium, which affects cell growth; at the same time, the state of the scaffold cells can be better observed, and it will not be affected by the position of the microscope lens when observing under the microscope, and different types of cells can be observed by directly turning it upside down.
[0045] The drawings are only for illustrative purposes and cannot be construed as limiting the present invention. To better illustrate the present embodiment, some parts of the drawings may be omitted, enlarged, or reduced, and do not represent the size of the actual product. For those skilled in the art, it is understandable that some well-known structures and their descriptions may be omitted in the drawings. The positional relationships described in the drawings are only for illustrative purposes and cannot be construed as limiting the present invention.
[0046] The same or similar reference numerals in the drawings of the embodiments of the present invention correspond to the same or similar parts; in the description of the present invention, it should be understood that if there are terms such as "upper", "lower", "left", "right", "long", "short", etc. indicating the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, it is 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. Therefore, the terms describing the positional relationship in the drawings are only used for exemplary explanations and cannot be understood as limitations on this patent. For ordinary technicians in this field, the specific meanings of the above terms can be understood according to the specific circumstances. In addition, the terms "first", "second", "third", etc. are only used for descriptive purposes and cannot be understood as indicating or implying relative importance. "Vertical" is not strictly vertical, but within the allowable error range. "Parallel" is not strictly parallel, but within the allowable error range.
[0047] In the description of this application, it is also necessary to explain that, unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense. For example, the specific relationship represented can be a fixed connection, a detachable connection, or an integral connection; it can be directly connected, or indirectly connected through an intermediate medium, or it can be the internal communication of two elements. For ordinary technicians in this field, they can understand the specific meanings of the above terms in this application according to specific circumstances.
[0048] Obviously, the above embodiments of the utility model are only examples for clearly explaining the utility model, and are not intended to limit the implementation methods of the utility model. For ordinary technicians in the relevant field, other different forms of changes or modifications can be made on the basis of the above description. It is not necessary and impossible to list all the implementation methods here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the utility model should be included in the protection scope of the claims of the utility model.
Claims
1. A common substrate double-layer cell culture device, characterized in that: It comprises a body and a fixing clip, wherein the body comprises a first shell, a second shell, and a culture dish, wherein the first shell and the second shell are hollow inside, a first surface of the culture dish abuts against one end of the first shell, and another surface of the culture dish abuts against one end of the second shell; The fixing clip comprises an annular belt body, and the fixing clip is sleeved on the outer side of the main body through the annular belt body to fix the first shell, the second shell and the culture dish.
2. The common substrate double-layer cell culture device according to claim 1, characterized in that: The first shell and the second shell are cylindrical.
3. The common substrate double-layer cell culture device according to claim 1, characterized in that: A surface of the first shell facing the second shell and a surface of the second shell facing the first shell are both provided with protrusions.
4. The common substrate double-layer cell culture device according to claim 1, characterized in that: The fixing clamp further comprises a fixing assembly, and the fixing assembly comprises fixing pieces, and the fixing pieces are passed through two ends of the annular belt body to fix the fixing clamp.
5. The common substrate double-layer cell culture device according to claim 4, characterized in that: The fixing assembly also includes a nut. The fixing member is a bolt. The bolt passes through two ends of the annular belt body. The nut is sleeved on the bolt. The nut is rotated to increase or decrease the diameter of the annular belt body.
6. The common substrate double-layer cell culture device according to claim 1, characterized in that: The fixing clip further comprises a supporting portion, and the supporting portion is arranged between the annular belt bodies to support the main body.
7. The common substrate double-layer cell culture device according to claim 1, characterized in that: A hydrophobic coating is provided on the inner walls of the first shell and the second shell.
8. A well plate, comprising the common substrate double-layer cell culture device according to any one of claims 1 to 7, characterized in that: The orifice plate is provided with a plurality of holes matched with the common-base double-layer cell culture device, and the common-base double-layer cell culture device is arranged in the holes.
9. The orifice plate according to claim 8, characterized in that A plurality of covers for closing the holes are arranged on the holes.
10. An incubator, characterized in that: It comprises a box body and the orifice plate according to claim 9, wherein the orifice plate is arranged in the box body.