A tcr-t cytotoxicity test microfluidic chip

Abstract

The utility model discloses a TCR -T cytotoxicity test micro -fluidic chip relates to micro -fluidic chip field, the utility model discloses a chip, the chip includes the chip, the chip includes the main part, be provided with micro -flow pass in the main part, the one side of main part is provided with the hose joint, the one side of hose joint is provided with the analog mechanism for the gas injection micro -flow pass, through set up the analog mechanism for the gas injection micro -flow pass, wherein analog mechanism includes the joint of screwing with hose joint, the hose of connecting joint, the gas pipe plug of the installation slot of end portion is equipped with, and the rubber gas valve of installation slot is used for the gas injection needle tube puncture, and the inside of hose joint is equipped with the breathable water -resisting membrane, solved the chip only can rely on external large -scale equipment adjustment environment gas, can not realize local dynamic gas concentration control in the inside, and the technical problem of poor sealing, easy gas -liquid mixing or pollution when gas injection.

Description

Technical Field

[0001] This utility model relates to the field of microfluidic chips, specifically a microfluidic chip for TCR-T cytotoxicity testing. Background Technology

[0002] As an important means of tumor immunotherapy, the preclinical efficacy assessment of TCR-T cell therapy relies on cytotoxicity testing that precisely simulates the tumor microenvironment in vitro. Microfluidic chips, which can construct microenvironments that closely resemble in vivo physiological conditions, such as dynamic fluids and intercellular interactions, have become a key tool for TCR-T cytotoxicity testing.

[0003] Microfluidic chips used for TCR-T cell toxicity testing primarily focus on functions such as cell co-culture and dynamic fluid transport. Their core structure typically includes a main body, microchannels, and fluid interfaces to enable basic operations such as contact between TCR-T cells and tumor cells and culture medium exchange. However, existing chips can only rely on external large-scale equipment, such as hypoxic incubators, to regulate the ambient gas. They cannot achieve localized and dynamic gas concentration control within the chip, such as simulating the oxygen concentration gradient between the tumor core and periphery. This results in a significant discrepancy between the in vitro functional performance of TCR-T cells and the actual in vivo situation, affecting the reference value of the test results. Utility Model Content

[0004] Based on this, the purpose of this utility model is to provide a microfluidic chip for TCR-T cell toxicity testing, in order to solve the technical problem that existing microfluidic chips can only rely on external large-scale equipment to regulate the ambient gas as a whole, and cannot achieve local and dynamic gas concentration control inside the chip, resulting in a significant deviation between the in vitro functional performance of TCR-T cells and the actual situation in vivo, thus affecting the reference value of the test results.

[0005] To achieve the above objectives, the present invention provides the following technical solution: a microfluidic chip for TCR-T cell toxicity testing, comprising a chip, the chip comprising a main body, the main body having a microchannel disposed therein, a flexible tube interface being provided on one side of the main body, and a simulated mechanism for injecting gas into the microchannel being provided on one side of the flexible tube interface; The simulation mechanism includes a connector, which is screwed to a hose interface. A hose is provided at the other end of the connector. An air pipe plug is provided at the end of the hose away from the connector. An installation groove is provided on one side of the air pipe plug. A rubber air valve that is inserted into an external air injection needle is provided in the installation groove.

[0006] By adopting the above technical solution, a secure connection is achieved through screw-on joints, the flexible hose is adapted for gas transmission, and the mounting groove of the gas tube plug secures the rubber gas valve. The entire system forms a dedicated gas injection path, compatible with the injection needle and ensuring a tight seal. This solves the problem of inconvenient control caused by the lack of a dedicated gas injection structure and improves test stability.

[0007] Furthermore, the hose is made of medical-grade rubber, and the connector is made of stainless steel.

[0008] By adopting the above technical solution, the tubing is made of medical-grade rubber, which has good flexibility and biocompatibility, can be adapted to the dynamic connection of the simulation mechanism and avoids cell contamination; the connector is made of stainless steel, which is high in strength and wear-resistant, ensuring structural stability when screwed to the tubing interface and extending the overall service life of the chip.

[0009] Furthermore, one end of the microchannel is provided with an interface, and the top of the interface is provided with a threaded connector.

[0010] By adopting the above technical solution, the interface at one end of the microchannel, combined with the top threaded connector, can be securely connected to an external infusion device. The threaded structure enhances sealing performance and prevents leakage of cell reagents. Simultaneously, the reagent injection pathway is clearly defined, ensuring accurate delivery of samples such as TCR-T cells and tumor cells into the microchannel and improving operational convenience.

[0011] Furthermore, the hose interface is connected to the microchannel, and a breathable and water-resistant membrane is provided on the inner side of the hose interface.

[0012] By adopting the above technical solution, the connection between the hose interface and the microchannel ensures smooth gas transmission, and the inner breathable and water-proof membrane achieves gas-liquid isolation, allowing gas to penetrate into the microchannel while blocking liquid from entering the simulation mechanism, avoiding gas-liquid mixing and interference with the test, and also preventing liquid from contaminating the gas path structure.

[0013] Furthermore, the two sides of the main body are provided with anti-slip textures, and the anti-slip textures are composed of multiple particle dots.

[0014] By adopting the above technical solution, the anti-slip texture on both sides of the main body is composed of multiple granular dots, which increases the friction between the hand and the main body, preventing the chip from slipping or shifting during operation. This makes it easier for operators to hold the chip for installation, gas injection, or observation, improving operational safety and stability.

[0015] Furthermore, the rubber valve is disc-shaped, and the surface of the rubber valve away from the mounting groove is an arc-shaped protrusion, with a weak area reserved in the middle of the arc-shaped protrusion for the injection needle to puncture.

[0016] By adopting the above technical solution, the disc-shaped rubber valve is securely fixed in the mounting groove, the arc-shaped protrusion facilitates the positioning of the injection needle, and the weak central area reduces the difficulty of puncture. The elasticity of the rubber material allows it to close automatically after puncture, preventing gas leakage and external contamination, adapting to multiple injection needs, and improving operational convenience.

[0017] Furthermore, the microchannel extends continuously in a serpentine shape within the main body, and the width of the microchannel remains consistent along the fluid flow direction. One end of the microchannel is connected to the fluid interface on the main body, and the other end is connected to the preset reaction area.

[0018] By adopting the above technical solution, the microchannels extend continuously in a serpentine shape with a consistent width, which stabilizes the fluid flow, avoids turbulence, and ensures uniform cell distribution. One end connects to the fluid interface, and the other end connects to the reaction area, clearly defining the reagent delivery pathway and ensuring that cells and reagents are accurately delivered to the reaction area, thereby improving the consistency of the test.

[0019] Furthermore, when the connector of the simulation mechanism is screwed into the hose interface, the outer wall of the connector is flush with the surface of the main body, and a sealing gasket is provided between the end of the connector and the inner wall of the hose interface.

[0020] By adopting the above technical solution, the connector, after being screwed in, is flush with the surface of the main body, avoiding damage from collisions to the protruding structure; the end sealing gasket enhances the seal with the inner wall of the hose interface, preventing gas leakage. Together, these two features improve the reliability of the connection between the analog mechanism and the chip, extending the structure's service life.

[0021] In summary, the present invention has the following main advantages: This invention solves the technical problems of existing chips that rely solely on external large-scale equipment to regulate ambient gas, cannot achieve local dynamic gas concentration control internally, and suffer from poor sealing, gas-liquid mixing, or contamination during gas injection by setting up a simulation mechanism for injecting gas into microchannels. The simulation mechanism includes a connector screwed to a hose interface, a hose connecting the connector, a gas tube plug with an installation groove at the end, and a rubber gas valve in the installation groove for puncturing the injection needle. Furthermore, a breathable and water-proof membrane is provided inside the hose interface. Attached Figure Description

[0022] Figure 1 This is a three-dimensional structural diagram of the present invention; Figure 2 This is a partial three-dimensional structural diagram of the simulation mechanism of this utility model; Figure 3 This utility model Figure 1 Enlarged structural diagram at point A; Figure 4 This utility model Figure 2 A magnified structural diagram at point B in the middle.

[0023] In the diagram: 1. Chip; 101. Main body; 102. Microchannel; 103. Interface; 104. Threaded connector; 105. Anti-slip texture; 106. Hose interface; 107. Breathable and water-resistant membrane; 2. Simulation mechanism; 201. Connector; 202. Hose; 203. Air pipe plug; 204. Mounting slot; 205. Rubber air valve. Detailed Implementation

[0024] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0025] A microfluidic chip for TCR-T cytotoxicity testing, such as Figure 1-4 As shown, the chip includes a chip 1, which includes a main body 101. A microchannel 102 is provided inside the main body 101. A hose interface 106 is provided on one side of the main body 101. A simulation mechanism 2 for injecting gas into the microchannel 102 is provided on one side of the hose interface 106. The simulation mechanism 2 includes a connector 201, which is screwed to a hose interface 106. A hose 202 is located at the other end of the connector 201. An air plug 203 is located at the end of the hose 202 furthest from the connector 201. A mounting groove 204 is provided on one side of the air plug 203, and a rubber valve 205, which connects to an external gas injection needle, is installed within the mounting groove 204. The main body 101 of the chip 1 provides the mounting base, the microchannel 102 provides a channel for cell reactions and fluid transport, and the hose interface 106 enables docking with the simulation mechanism 2. The screw connection between the connector 201 and the hose interface 106 enhances connection stability. The hose 202 provides flexible adaptation for gas transport. The mounting groove 204 of the air plug 203 provides a fixed space for the rubber valve 205, which is adapted to an external gas injection needle. A dedicated gas injection system was constructed to allow for precise injection of gas into the microchannel 102 without relying on external large equipment. At the same time, the various components work together to ensure a gas injection seal, preventing contamination and leakage, and improving the accuracy and convenience of testing.

[0026] See Figure 1 , Figure 2 , Figure 3 , Figure 4The tubing 202 is made of medical-grade rubber, and the connector 201 is made of stainless steel. The tubing 202 of the simulation mechanism 2 is made of medical-grade rubber, whose flexibility allows for adjustment of the connection angle between the connector 201 and the endotracheal plug 203, meeting the layout requirements of different gas injection scenarios. Simultaneously, the medical-grade material is non-toxic, preventing contamination when in contact with cell reagents and ensuring the activity of test samples. The connector 201 is made of stainless steel, possessing excellent structural strength and wear resistance. It is not easily deformed during repeated screwing and assembly with the tubing interface 106 and can withstand the pressure during gas injection, preventing gas leakage due to connector damage. Combined with the material properties of the tubing 202, this improves the reliability and service life of the simulation mechanism 2.

[0027] See Figure 1 , Figure 2 , Figure 3 , Figure 4 One end of the microfluidic channel 102 is provided with an interface 103, and the top of the interface 103 is provided with a threaded connector 104. The interface 103 at one end of the microfluidic channel 102 provides a dedicated channel for reagent injection. The threaded connector 104 at the top of the interface 103 adopts a threaded connection method, which can be precisely adapted to the interface of the external infusion device. The tight engagement of the thread effectively enhances the connection sealing performance, preventing cell reagent leakage or the entry of outside air into the microfluidic channel 102 during injection, thus ensuring sample concentration and activity. Through this structure, operators can quickly complete the docking of the external infusion device with the chip 1, enabling the precise injection of mixed systems such as TCR-T cells, tumor cells, and culture medium into the microfluidic channel 102. The clear pathway design avoids reagent injection confusion and improves operational efficiency and testing stability.

[0028] See Figure 1 , Figure 2 , Figure 3 , Figure 4 The flexible hose interface 106 is connected to the microchannel 102, and a breathable and water-resistant membrane 107 is provided on the inner side of the flexible hose interface 106. This connection between the flexible hose interface 106 and the microchannel 102 creates a dedicated gas transmission path from the simulation mechanism 2 to the microchannel 102, ensuring that the injected gas can smoothly reach the reaction area. The breathable and water-resistant membrane 107 on the inner side of the flexible hose interface 106 has both gas permeability and liquid barrier properties. Gas can permeate through the membrane into the microchannel 102 to regulate the environment, while liquids such as cell reagents in the microchannel 102 are blocked and cannot enter the flexible hose interface 106 or the gas path of the simulation mechanism 2. This design avoids cell distribution disorder caused by gas-liquid mixing, ensuring a stable testing environment, and prevents liquid from entering the gas path and causing contamination or blockage of the simulation mechanism 2, thus improving the accuracy of gas regulation and structural cleanliness.

[0029] See Figure 1 , Figure 2 , Figure 3 , Figure 4 The main body 101 has anti-slip textures 105 on both sides, and these textures 105 are composed of multiple granular dots. The distribution of these granular dots effectively increases the contact friction between the operator's hand and the surface of the main body 101. This prevents the chip 1 from shifting due to hand slippage during operations such as assembling the simulation mechanism 2, puncturing with a gas injection needle, or microscopic observation. Especially when precisely docking with external devices or observing the cell state in the reaction area, the anti-slip design improves operational stability and avoids interface docking errors, gas injection deviations, or observation errors caused by chip shifting. At the same time, the granular dot structure is simple and does not affect the overall structural strength and transparent observation performance of the main body 101, thus balancing practicality and functionality.

[0030] See Figure 1 , Figure 2 , Figure 3 , Figure 4 The rubber valve 205 is disc-shaped, with an arc-shaped protrusion on the side of the valve away from the mounting groove 204. A weak area is reserved in the center of this arc-shaped protrusion for the injection needle to puncture. The disc-shaped design of the rubber valve 205 allows for precise contact with the mounting groove 204 of the air tube plug 203, ensuring a secure fixation and preventing valve displacement during injection. The arc-shaped protrusion on the side of the rubber valve 205 away from the mounting groove 204 provides clear puncture guidance for the external injection needle, facilitating quick alignment by the operator. The reserved weak area in the center reduces resistance during needle puncture, improving puncture convenience. The rubber material has good elasticity; after the injection needle is removed, the puncture hole in the weak area automatically closes, achieving a gas path seal and preventing internal gas leakage or contamination from external impurities. This structure can withstand multiple punctures, adapting to different gas control needs and improving operational flexibility and sealing.

[0031] See Figure 1 , Figure 2 , Figure 3 , Figure 4The microchannel 102 extends continuously in a serpentine pattern within the main body 101, with its width remaining consistent along the fluid flow direction. One end of the microchannel 102 connects to a fluid interface on the main body 101, while the other end connects to a pre-defined reaction area. This continuous serpentine design, combined with a consistent width along the fluid flow direction, effectively optimizes fluid flow, preventing turbulence caused by abrupt path changes or width variations. This ensures a uniform distribution of the TCR-T cell and tumor cell mixture during flow, preventing cell accumulation or uneven dispersion. The connection between the microchannel 102 and the fluid interface on the main body 101, along with the pre-defined reaction area, establishes a complete pathway from reagent injection to reaction. This ensures that injected cells and reagents are smoothly and accurately delivered to the reaction area, avoiding abnormal reaction environments caused by pathway disruptions and improving the consistency of conditions and the reliability of results between different test samples.

[0032] See Figure 1 , Figure 2 , Figure 3 , Figure 4 When the connector 201 of the simulation mechanism 2 is screwed onto the hose interface 106, the outer wall of the connector 201 is flush with the surface of the main body 101, and a sealing gasket is provided between the end of the connector 201 and the inner wall of the hose interface 106. This flush design prevents the connector 201 from protruding from the main body surface and forming a collision-prone structure, thus preventing deformation or breakage due to collisions during operation or storage, and ensuring the integrity of the connection structure. The sealing gasket between the end of the connector 201 and the inner wall of the hose interface 106 is squeezed and filled during tightening, significantly enhancing the sealing performance between the two, preventing gas leakage from the interface gap, and ensuring that all injected gas enters the microchannel 102 through the hose interface 106. The flush design and the sealing gasket not only improve the safety and durability of the structure but also ensure the accuracy of gas control, enhancing the overall structural reliability.

[0033] The implementation principle of this embodiment is as follows: First, the chip 1 and the simulation mechanism 2 are assembled. The connector 201 of the simulation mechanism 2 is screwed and fixed to the hose interface 106 on the main body 101. At this time, the outer wall of the connector 201 is flush with the surface of the main body 101, and the sealing gasket between the end of the connector 201 and the inner wall of the hose interface 106 is pressed to achieve a sealed connection between the two. At the same time, the breathable and water-proof membrane 107 preset on the inner side of the hose interface 106 can block liquid from passing through and allow gas to permeate, laying the foundation for subsequent gas-liquid isolation. The connector 201 is made of stainless steel to ensure structural strength, and the hose 202 is made of medical rubber to ensure flexibility and biocompatibility. Subsequently, the cells and reagents required for the experiment are injected into the chip 1: an external infusion device is connected through the threaded connector 104 at the top of the interface 103 at one end of the microchannel 102, and the mixture of TCR-T cells, tumor cells and culture medium is injected into the microchannel 102; since the microchannel 102 extends continuously in a serpentine shape within the main body 101 and its width remains consistent along the direction of fluid flow, the fluid can form a stable flow state during the flow process, avoiding uneven cell distribution caused by turbulence, and the mixture can flow smoothly through the microchannel 102 and finally enter the preset reaction area to complete the spreading; Next, the target gas is injected into the microchannel 102 through the simulation mechanism 2 to regulate the reaction environment: the external gas injection needle is aligned with the rubber valve 205 in the mounting slot 204 of the endotracheal plug 203, and the weak area in the middle of the arc-shaped protrusion of the rubber valve 205 is punctured; after the target gas is injected through the gas injection needle, it passes through the rubber valve 205, endotracheal plug 203, tubing 202 and connector 201 in sequence into the tubing interface 106, and then permeates into the microchannel 102 through the breathable and water-proof membrane 107, contacting the cell system in the reaction area to construct the target gas environment; after the gas injection is completed, the gas injection needle is pulled out, and the elasticity of the rubber valve 205 can automatically close the puncture site to prevent gas leakage or external contamination; Throughout the experiment, the operator can hold the chip 1 stably using the anti-slip texture 105 composed of multiple particles on both sides of the main body 101, preventing slippage and chip 1 from shifting during operation; the interaction between TCR-T cells and tumor cells in the reaction area can be observed through the transparent main body 101 of the chip 1; the connection design between the microchannel 102 and the reaction area ensures nutrient supply and metabolic product discharge; the sealing and permeability design of the simulation mechanism 2 enables precise gas control; and the synergistic effect of each structure completes the TCR-T cell toxicity test.

[0034] Although embodiments of the present invention have been shown and described, these specific embodiments are merely explanations of the present invention and are not intended to limit the invention. The specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. After reading this specification, those skilled in the art may make modifications, substitutions, and variations to the embodiments as needed without departing from the principles and spirit of the present invention, provided that such modifications, substitutions, and variations are within the scope of the claims of the present invention and are protected by patent law.

Claims

1. A microfluidic chip for TCR-T cell toxicity testing, characterized in that: Includes a chip (1), the chip (1) includes a body (101), the body (101) is provided with a microchannel (102), a hose interface (106) is provided on one side of the body (101), and a simulation mechanism (2) for injecting gas into the microchannel (102) is provided on one side of the hose interface (106). The simulation mechanism (2) includes a connector (201), which is screwed to a hose interface (106). A hose (202) is provided at the other end of the connector (201). An air pipe plug (203) is provided at the end of the hose (202) away from the connector (201). An installation groove (204) is provided on one side of the air pipe plug (203). A rubber air valve (205) that is inserted into an external air injection needle is provided in the installation groove (204).

2. The TCR-T cytotoxicity test microfluidic chip according to claim 1, wherein: The hose (202) is made of medical rubber, and the connector (201) is made of stainless steel.

3. The microfluidic chip for TCR-T cell toxicity testing according to claim 1, characterized in that: One end of the microchannel (102) is provided with an interface (103), and the top of the interface (103) is provided with a threaded connector (104).

4. The TCR-T cytotoxicity test microfluidic chip of claim 1, wherein: The hose interface (106) is connected to the microchannel (102), and a breathable and water-proof membrane (107) is provided on the inner side of the hose interface (106).

5. The TCR-T cytotoxicity test microfluidic chip of claim 1, wherein: The main body (101) has anti-slip textures (105) on both sides, and the anti-slip textures (105) are composed of multiple particle dots.

6. The TCR-T cytotoxicity test microfluidic chip of claim 1, wherein: The rubber valve (205) is in the shape of a disc, and the surface of the rubber valve (205) away from the mounting groove (204) is an arc-shaped protrusion, with a weak area reserved in the middle of the arc-shaped protrusion for the injection needle to puncture.

7. The TCR-T cytotoxicity test microfluidic chip of claim 1, wherein: The microchannel (102) extends continuously in a serpentine shape within the main body (101), and the width of the microchannel (102) remains consistent along the fluid flow direction. One end of the microchannel (102) is connected to the fluid interface on the main body (101), and the other end is connected to the preset reaction area.

8. The TCR-T cytotoxicity test microfluidic chip of claim 1, wherein: When the connector (201) of the simulation mechanism (2) is screwed into the hose interface (106), the outer wall of the connector (201) is flush with the surface of the main body (101), and a sealing gasket is provided between the end of the connector (201) and the inner wall of the hose interface (106).

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