Simple open type tension organ chip and pouring mold thereof
By designing simple open tension organ chips and corresponding casting molds, the problem of difficulty in simulating multiple mechanical microenvironments in the existing technology is solved, and flexible regulation of diversified mechanical stimulation and drug stimulation is achieved, which improves the manufacturing efficiency of the organ chip and Experimental efficacy.
Patent Information
- Application Number
- CN202421050450.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-05-15
- Publication Date
- 2025-01-07
- Estimated Expiration
- 2034-05-15
AI Technical Summary
Existing organ chips are difficult to simulate many different mechanical microenvironments and cannot meet the reproduction needs of complex biomechanical environments, especially in drug screening and disease simulation.
A simple open tension organ chip was designed, including cell culture structure, airbags, negative pressure channels and nutrient channels. Combined with casting molds, it can achieve flexible regulation of diversified mechanical stimulation and drug stimulation, and support the manufacturing and experimental operation of organ chips. .
The chip can simulate diverse mechanical stimulation, improve the efficiency of drug screening and disease simulation, meet the needs of organ chip manufacturing, provide flexible experimental conditions, and support multi-faceted research needs.
Smart Images
Figure CN222313187U_ABST
Abstract
Description
Technical Field
[0001] The utility model belongs to the field of biotechnology, and particularly relates to a simple open tension organ chip and a casting mold thereof. Background Art
[0002] Organ chips are biotechnology that simulates and reproduces the microenvironments of various living cells, tissues, and organs in microfluidic chips; this technology can reproduce the mechanisms of action in vivo in vitro. Different types of organ chips, such as intestine chips, heart chips, lung chips, liver chips, brain chips, and tumor chips, have been born and are widely used in drug screening, disease simulation, personalized medicine, and toxicity testing. In these organ chips, it is particularly important to simulate and reproduce the personalized biomechanical environment of cells, such as simulating the periodic contraction and relaxation of alveoli, blood flow in blood vessels, regular beating of the heart, the forward and backward movement of fluid in bone pores, and the peripheral environment of tumor cell migration. Utility Model Content
[0003] The purpose of the utility model is to meet the ability to simulate a variety of different mechanical microenvironments and provide a simple open tension organ chip.
[0004] Another purpose of the utility model is to meet the needs of organ chip manufacturing and provide a simple open tension organ chip casting mold.
[0005] The technical solution provided by the utility model is:
[0006] A simple open tension organ chip, comprising:
[0007] A cell culture structure, which is a rectangular parallelepiped;
[0008] The interior of the cell culture structure comprises:
[0009] A cell culture chamber, which is a rectangular through hole in the cell culture structure;
[0010] A nutrient inlet channel and a nutrient outlet channel, which are respectively arranged on both sides of the wide side of the cell culture chamber; one end of the nutrient inlet channel and one end of the nutrient outlet channel are respectively connected to the cell culture chamber;
[0011] Two air bags are respectively arranged on both sides of the long side of the cell culture chamber;
[0012] an airbag channel, one end of which is connected to the two airbags respectively;
[0013] A negative pressure channel, which is arranged around the cell culture chamber, the nutrient inlet channel, the nutrient outlet channel, the two air bags and the air bag channel;
[0014] A cell culture cover, which covers the top of the cell culture structure;
[0015] A cell attachment membrane is attached to the bottom of the cell culture structure.
[0016] Preferably, the organ chip further comprises: a negative pressure exhaust port, a nutrient inlet port, a nutrient outlet port and an air bag interface, all of which are circular through holes; the negative pressure exhaust port is arranged at one end outside the negative pressure channel and is connected to the negative pressure channel; the nutrient inlet port, the nutrient outlet port and the air bag interface are respectively connected to the other ends of the nutrient inlet channel, the nutrient outlet channel and the air bag channel.
[0017] Preferably, the cell culture cover comprises:
[0018] A main body, which is a rectangular parallelepiped;
[0019] A rectangular convex portion connected to the bottom of the main body; the rectangular convex portion is completely embedded in the cell culture chamber;
[0020] A plurality of cylindrical protrusions are connected to the bottom of the main body; the plurality of cylindrical protrusions correspond one by one to the negative pressure suction port, the nutrient inlet port, the nutrient outlet port and the air bag interface respectively.
[0021] Preferably, the cell attachment membrane is an elastic PDMS membrane.
[0022] Preferably, the thickness of the PDMS film does not exceed 0.18 mm.
[0023] Preferably, a microstructure is disposed on the upper surface of the cell attachment membrane; and the microstructure is disposed directly below the cell culture chamber.
[0024] Preferably, a casting mold for a simple open tension organ chip is used for casting a cell culture structure of a simple open tension organ chip, and the casting mold comprises:
[0025] The casting mold top cover is in the shape of a cuboid, and a rectangular through hole is opened in the middle of the casting mold top cover;
[0026] The casting frame module is a rectangular shell that penetrates from top to bottom; the casting mold top cover is covered on the top of the casting frame module;
[0027] The culture chip space structure mold module has a base that is a rectangular parallelepiped and matches and fits with the casting frame module. There is a relief pattern corresponding to the cell culture structure in the middle of the culture chip space structure mold module. The part of the relief pattern corresponding to the cell culture bin is the culture bin module. The culture bin module is a rectangular parallelepiped. The top of the culture bin module is completely matched and fits with the rectangular through hole in the top cover of the casting mold. The casting frame module is arranged on the outside of the culture chip space structure mold module.
[0028] The beneficial effects of the utility model are:
[0029] The simple open tension organ chip provided by the utility model can carry out diversified mechanical stimulation, and the combination of mechanical stimulation and drug stimulation is flexible and adjustable.
[0030] The simple open tension organ chip casting mold provided by the utility model has a simple structure and is easy to operate, thus meeting the requirements of organ chip manufacturing and improving the efficiency of organ chip manufacturing. BRIEF DESCRIPTION OF THE DRAWINGS
[0031] Figure 1 This is a schematic diagram of the cell culture structure described in the utility model.
[0032] Figure 2 This is a schematic diagram of the cell attachment membrane described in the present utility model.
[0033] Figure 3 This is a schematic diagram of the cell attachment membrane cylinder array described in the present utility model.
[0034] Figure 4 This is a schematic diagram of the organ chip assembly described in the present invention.
[0035] Figure 5 This is a schematic diagram of the casting mold of the organ chip described in the present invention.
[0036] Figure 6 Schematic diagram of the adhesive film pressing structure. DETAILED DESCRIPTION
[0037] The present invention will be further described in detail below in conjunction with the accompanying drawings so that those skilled in the art can implement the invention with reference to the description.
[0038] like Figure 1 As shown, the cell culture structure 120 has a rectangular shape;
[0039] The interior of the cell culture structure 120 includes:
[0040] The cell culture chamber 121 is a rectangular through hole in the cell culture structure 120;
[0041] The nutrient inlet channel 123 and the nutrient outlet channel 124 are respectively arranged on both sides of the wide side of the cell culture chamber 121; one end of the nutrient inlet channel 123 and one end of the nutrient outlet channel 124 are respectively connected to the cell culture chamber 121;
[0042] Two air bags 125, which are respectively arranged on both sides of the long side of the cell culture chamber 121;
[0043] An airbag channel 126, one end of which is connected to the two airbags 125 respectively;
[0044] A negative pressure channel 122 is arranged on the outside near the bottom of the cell culture structure 120. The negative pressure channel 122 is arranged around the cell culture chamber 121, the nutrient inlet channel 123, the nutrient outlet channel 124, the two air bags 125 and the air bag channel 126. The negative pressure channel 122 is rectangular.
[0045] After the chip is assembled, holes are punched at positions corresponding to the negative pressure channel 122, the nutrient inlet channel 123, the nutrient outlet channel 124 and the air bag channel 126 to form four visible circular through holes 140 that penetrate the cell culture structure 120 from top to bottom, which serve as interfaces for each channel. The four circular through holes 140 are respectively a negative pressure air extraction port 141, a nutrient inlet port 142, a nutrient outlet port 143 and an air bag interface 144; the negative pressure air extraction port 141 is arranged at one end outside the negative pressure channel 122 and is connected to the negative pressure channel 122; the nutrient inlet port 142, the nutrient outlet port 143 and the air bag interface 144 are respectively connected to the other ends of the corresponding nutrient inlet channel 123, the nutrient outlet channel 124 and the air bag channel 126.
[0046] In order to meet the needs of use in a variety of environments, the cell culture chamber 121 has four different sizes and combination versions, and the functions are consistent with the above introduction.
[0047] like Figure 2 As shown, the cell attachment membrane 130 is an elastic PDMS membrane with a thickness not exceeding 0.18 mm, which is suitable for confocal oil-immersion fluorescence microscopy imaging and real-time observation; the cell attachment membrane 130 is adsorbed on the bottom of the cell culture structure 120; under the traction of the airbag 125, the cell attachment membrane 130 can undergo a certain elastic deformation.
[0048] The upper surface of the cell attachment membrane 130 is a smooth surface or a surface provided with a microstructure 131 ; the microstructure 131 is arranged directly below the cell culture chamber 121 .
[0049] The material selection of the cell attachment membrane 130 is diverse, and can be nitrocellulose membrane, nylon membrane, glass membrane, etc. The selected cell attachment membrane 130 should have a certain elasticity and should be able to have a certain elastic deformation under the traction of the airbag 125.
[0050] like Figure 3 As shown, the microstructure 131 is a micro-nano structure pattern or other microstructures, the micro-nano structure pattern is a micro-nano cylinder array, the micro-nano cylinder 132 has a diameter of 0.02 mm and a height of 0.04 mm, and the spacing between two adjacent micro-nano cylinders 132 is 0.06 mm; the other microstructures are grooves with a spacing of 1 μm and a depth of 0.2 μm, or soft nano pillars with a diameter of 700 nm, a height of 1500 nm, and a spacing of 800 nm.
[0051] Cells can be cultured in the micro-nano cylinders 132 , and a certain distance is left between two adjacent micro-nano cylinders 132 , so as to facilitate observation of the influence between cells when the cells are stretched.
[0052] like Figure 4 As shown, the utility model provides a simple open tension organ chip, including: a cell culture cover 110, which is covered on the top of the cell culture structure 120; the cell culture cover 110 includes: a main body 111, a rectangular convex part 112 and a plurality of cylindrical convex parts 113; the main body 111 is a rectangular prism; the rectangular convex part 112 and the cylindrical convex part 113 are connected to the bottom of the main body 111; the rectangular convex part 112 is completely embedded with the cell culture chamber 121; the number of the cylindrical convex parts 113 is four, and the cylindrical convex parts 113 are respectively embedded with the negative pressure exhaust port 141, the nutrient inlet port 142, the nutrient outlet port 143 and the airbag interface 144.
[0053] The negative pressure suction port 141 is connected to negative pressure to provide suction to tighten the cell attachment layer 130. The airbag interface 144 is connected to negative pressure to allow gas to control the airbag 125 through the gas channel, causing it to deform and drive the cell attachment membrane 130 to deform, giving the cells a pulling force.
[0054] Nutrients enter through the nutrient inlet port 142 at the top of the nutrient inlet channel 123 to provide nutrients for the cells in the cell culture chamber 121, and waste liquid such as metabolic products is taken away through the nutrient outlet channel 124 and the nutrient outlet 143; at the same time, negative pressure can be applied at the nutrient outlet 143 when adding liquid to speed up the nutrient supply.
[0055] When the chip is cultured in an incubator, the cell culture cover 110 is covered to provide a more sterile environment for the cell culture chamber 121 and each channel.
[0056] First, the cell attachment membrane 130 is attached to the bottom of the cell culture structure 120 through the negative pressure channel 122 to form a complete chip, and the cells to be observed are placed in the cell culture chamber 121 for culture, sample addition, reaction and observation. By applying tension to the airbag 125, the cell attachment layer 130 is deformed, driving the cells to deform, so as to facilitate the observation of the effects of the cells under different tensions and deformations. Then, nutrients are injected into the cell culture chamber 121 through the nutrient entry channel 123 to provide the cells with necessary nutrients. At the same time, there is also a nutrient discharge channel 124 at the other end of the cell culture chamber 121 to discharge waste liquid, thereby realizing the culture of cells and the observation of cells under the influence of different tensions.
[0057] There are various choices of chip materials, and transparent materials are generally chosen to facilitate observation of cells.
[0058] The experimental process is divided into three steps: the first step is to assemble the chip; the second step is to plant cells in the cell culture chamber and cover and seal the cell culture cover 110; the third step is to apply tension to conduct the experiment.
[0059] Step 1: All glass objects are immersed in sulfuric acid for 12 hours, and then taken out wearing gloves, and the remaining sulfuric acid is rinsed off with running water, immersed in 75% alcohol for several rinses, and then cleaned in distilled water, placed on a slide rack for drying and ready for use; silicone objects are cleaned with a special detergent; the cell attachment membrane 130 is prepared; the cell culture structure 120 and the cell attachment membrane 130 are assembled, and after the two are tightly fitted, the negative pressure channel 122 is negatively evacuated to prevent the two from separating and the sealing is better, so that liquid will not leak out during operations such as planting cells.
[0060] Step 2: Plant cells at the center of the cell culture chamber 121 and seal it with the cell culture cover 110. Inject the nutrient solution required for cell culture through the nutrient inlet channel 123. After completion, place the chip in a carbon dioxide constant temperature incubator for culture. Take it out for observation after a period of time. If the cell culture meets the required requirements, proceed to the third step; if the culture solution is insufficient, apply negative pressure to the nutrient outlet channel 124 to extract the original nutrient and other waste liquids, and re-inject nutrients through the nutrient inlet channel 123 for culture. At the same time, if cell drugs need to be added, they can also be added through the nutrient inlet channel 123.
[0061] Step 3: After the cell culture is completed, the seal is removed and the negative pressure exhaust port 141 is connected to negative pressure to make the cell attachment membrane 130 adhere more closely. Then, negative pressure is connected through the airbag channel 126, so that the gas can pass through the airbag channel 126 to control the airbag 125 to deform, thereby driving the third layer of the cell attachment membrane 130 to deform and applying tension to the cells, so as to observe the cell morphology and structure under different conditions.
[0062] like Figure 5 As shown, the utility model provides a simple open tension organ chip casting mold, which includes: a casting mold top cover 210, which is in the shape of a cuboid, and a rectangular through hole 211 is opened in the middle of the casting mold top cover 210; the casting mold top cover 210 is used to compact the gel chip to prevent gel from overflowing and control the maximum height of the gel chip to ensure that the height of the chip after the mold is consistent;
[0063] The casting frame module 220 is a rectangular shell that is connected from top to bottom and has a certain height, so that the chip can be molded according to the needs to obtain a chip of a certain height. The casting mold top cover 210 is covered on the top of the casting module 220.
[0064] The culture chip space structure mold module 230 has a base 231 that is a rectangular parallelepiped, and the base 231 is matched and embedded with the casting frame module 220. The middle of the culture chip space structure mold module 230 has a relief pattern corresponding to the cell culture structure 120, and the part of the relief pattern corresponding to the cell culture chamber 121 is the culture chamber module 232. The culture chamber module 232 is a rectangular parallelepiped, and the top of the culture chamber module 232 is completely matched and embedded with the rectangular through hole 211 in the casting mold top cover 210; the casting frame module 220 is arranged on the outside of the culture chip space structure mold module 230.
[0065] The culture chip space structure mold module 230 is combined with the casting frame module 220, and the two are completely embedded, so the gel will not leak out from the bottom, and the gel is poured into the casting frame module 220, and the gel is compacted with the casting mold top cover 210; because the culture chip space structure mold module 230 has a relief pattern corresponding to the cell culture structure 120, the gel will solidify into a gel chip with a corresponding space structure according to the relief pattern after the mold is poured. After the gel is solidified, the casting mold top cover 210, the casting frame module 220, and the culture chip space structure mold module 230 are separated from the gel to obtain a chip with a corresponding space structure.
[0066] like Figure 6As shown, the attachment film pressing structure 310 is a rectangular structure with protrusions on all sides and a depression in the middle; the depression 311 of the attachment film pressing structure 310 is consistent in size with the cell culture structure 120 and can be completely fitted, and is used to assemble the cell culture structure 120 and the cell attachment film 130. The cell attachment film 130 is placed in the depression 311 of the attachment film pressing structure, and the cell culture structure 120 after the reverse molding is placed in the depression 311. After the cell culture structure 120 and the cell attachment film 130 are completely fitted, the chip fitted with the cell attachment film 130 can be directly taken out as a whole without damaging the chip structure.
[0067] Organ chip casting process:
[0068] Step 1: Prepare the chip gel. Prepare the chip gel of the materials you need in advance. The gel needs to be prepared and used immediately, otherwise the gel will solidify if it is used for a long time.
[0069] Step 2: Assemble the casting frame module 220 and the culture chip space structure mold module 230, and make them fit together completely. Pour the prepared gel into the casting frame module 220, and pour the gel according to the chip height you need.
[0070] Step 3: Cover the casting mold top cover 210 on the casting frame module 220 with the gel poured in it, and completely fit the rectangular through hole 211 in the casting mold top cover 210 with the culture chamber module 232 of the culture chip space structure die module. Press the bubbles in the gel lightly with your hands, and let the casting mold top cover 210 determine the height; put the assembled casting mold into the relevant environment for solidification, and remove the template after solidification for 24 hours before proceeding to the next step
[0071] Step 4: Place the cell attachment membrane 130 into the groove 311 of the attachment membrane pressing structure 310, and place the solidified cell culture structure 120 on top of the cell attachment membrane 130. After they are tightly attached, they can be taken out to form a complete chip structure, and experiments can be carried out.
[0072] The simple open tension organ chip provided by the utility model provides a variety of mechanical stimulation: the chip can simulate a variety of mechanical microenvironments, such as tensions of different intensities and frequencies, and different modes of fluid shear force or drug stimulation; the combination of mechanical stimulation and drug stimulation in the chip is flexible and adjustable: allowing users to adjust the combination of mechanical stimulation and drug stimulation according to needs, meeting various research needs, and helping to discover scientific problems; the chip can be open for cell culture, which is simple to operate and convenient for collecting cells for quantitative detection and analysis of nucleic acids and proteins; the chip meets the needs of confocal real-time imaging, provides high-definition images, and helps to accurately analyze cell responses and draw accurate conclusions.
[0073] The simple open tension organ chip casting mold provided by the utility model has a simple structure and is easy to operate, which meets the needs of organ chip manufacturing and improves the efficiency of organ chip manufacturing.
[0074] Although the implementation scheme of the utility model has been disclosed as above, it is not limited to the applications listed in the specification and implementation modes. It can be fully applied to various fields suitable for the utility model. For those familiar with the art, additional modifications can be easily implemented. Therefore, without departing from the general concept defined by the claims and the scope of equivalents, the utility model is not limited to the specific details and the illustrations shown and described herein.
Claims
1. A simple open tension organ chip, characterized in that: include: A cell culture structure, which is a rectangular parallelepiped; The interior of the cell culture structure comprises: A cell culture chamber, which is a rectangular through hole in the cell culture structure; A nutrient inlet channel and a nutrient outlet channel, which are respectively arranged on both sides of the wide side of the cell culture chamber; one end of the nutrient inlet channel and one end of the nutrient outlet channel are respectively connected to the cell culture chamber; Two air bags are respectively arranged on both sides of the long side of the cell culture chamber; an airbag channel, one end of which is connected to the two airbags respectively; a negative pressure channel, which is arranged around the cell culture chamber, the nutrient inlet channel, the nutrient outlet channel, the two air bags and the air bag channel; A cell culture cover, which covers the top of the cell culture structure; A cell attachment membrane is attached to the bottom of the cell culture structure.
2. The simple open tension organ chip according to claim 1, characterized in that: The organ chip also includes: a negative pressure exhaust port, a nutrient inlet port, a nutrient outlet port and an air bag interface, all of which are circular through holes; the negative pressure exhaust port is arranged at one end outside the negative pressure channel and is connected to the negative pressure channel; the nutrient inlet port, the nutrient outlet port and the air bag interface are respectively connected to the other end of the nutrient inlet channel, the nutrient outlet channel and the air bag channel.
3. The simplified open tension organ chip according to claim 2, characterized in that: The cell culture cover comprises: A main body, which is a rectangular parallelepiped; A rectangular convex portion connected to the bottom of the main body; the rectangular convex portion is completely embedded in the cell culture chamber; A plurality of cylindrical protrusions are connected to the bottom of the main body; the plurality of cylindrical protrusions correspond one by one to the negative pressure suction port, the nutrient inlet port, the nutrient outlet port and the air bag interface respectively.
4. The simplified open tension organ chip according to claim 1, characterized in that: The cell attachment membrane is an elastic PDMS membrane.
5. The simplified open tension organ chip according to claim 4, characterized in that: The thickness of the PDMS film does not exceed 0.18 mm.
6. The simplified open tension organ chip according to claim 4, characterized in that: A microstructure is arranged on the upper surface of the cell attachment membrane; the microstructure is arranged directly below the cell culture chamber.
7. A casting mold for a simple open tension organ chip, used for casting the cell culture structure of the simple open tension organ chip as described in any one of claims 1 to 6, characterized in that: Casting mold includes: The casting mold top cover is in the shape of a cuboid, and a rectangular through hole is opened in the middle of the casting mold top cover; The casting frame module is a rectangular shell that penetrates from top to bottom; the casting mold top cover is covered on the top of the casting frame module; The culture chip space structure mold module has a base that is a rectangular parallelepiped and matches and fits with the casting frame module. There is a relief pattern corresponding to the cell culture structure in the middle of the culture chip space structure mold module. The part of the relief pattern corresponding to the cell culture bin is the culture bin module. The culture bin module is a rectangular parallelepiped. The top of the culture bin module is completely matched and fits with the rectangular through hole in the top cover of the casting mold. The casting frame module is arranged on the outside of the culture chip space structure mold module.