Micro-fluidic chip
By designing the flow channels and three-dimensional culture chambers of microfluidic chips, combined with solenoid valves and biosensors, the challenge of personalized medicine for lung cancer has been solved, enabling multi-drug evaluation and the formulation of precise treatment plans, thereby improving the effectiveness and safety of lung cancer treatment.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-03
- Publication Date
- 2026-03-24
AI Technical Summary
The existing technologies for lung cancer suffer from problems such as drug resistance to targeted therapy and low response rates to immunotherapy, and lack effective efficacy prediction indicators, making personalized medication difficult.
Design a microfluidic chip containing multiple channels and a three-dimensional culture chamber. Control drug delivery via solenoid valves and combine it with biosensors to detect cytokines, simulating the immune microenvironment of lung cancer, to achieve high-throughput drug evaluation and personalized medication regimen development.
It enables high-throughput evaluation of multiple drug regimens, simulates the real human body environment, precisely controls dosage, reduces cell damage, provides reliable drug sensitivity test results, and guides personalized treatment.
Smart Images
Figure CN224031014U_ABST
Abstract
Description
Technical Field
[0001] This utility model relates to the field of biological detection technology, and in particular to a microfluidic chip. Background Technology
[0002] Lung cancer, as one of the leading causes of cancer-related deaths worldwide, poses a serious challenge to global public health. This is particularly true in my country, where its high incidence and mortality rates pose a significant threat to public health. Therefore, the prevention and treatment of lung cancer is especially urgent and important, as it not only affects individual quality of life but also directly impacts the overall health level of society and the allocation of medical resources.
[0003] With advancements in medical technology, lung cancer treatment has undergone a significant transformation, evolving from traditional surgery and chemotherapy to the more recent emergence of targeted therapy and immunotherapy. Among these, targeted therapy and immunotherapy, represented by PD-1 / PD-L1 inhibitors, have significantly improved treatment outcomes and quality of life for lung cancer patients due to their high efficacy and low toxicity, bringing new hope to many. Targeted therapy precisely targets specific tumor-related genes or proteins, reducing damage to normal cells; while immunotherapy modulates the patient's own immune system, stimulating its ability to fight tumors, thus achieving a breakthrough in treatment methods.
[0004] However, despite the significant achievements of these novel therapies, two major challenges remain in clinical practice: first, the problem of drug resistance in targeted therapies, where some patients experience a decrease in tumor sensitivity to drugs over time, leading to weakened treatment efficacy; and second, the low response rate of immunotherapy, with significant differences in efficacy among different patients, and some patients even showing no response at all. The root of these two problems lies in the high heterogeneity of lung cancer, meaning that there are significant differences in the tumor microenvironment (including tumor cells themselves, surrounding blood vessels, immune cells, cytokines, etc.) among different patients. This difference leads to uncertainty in treatment response and a lack of reliable predictive indicators of efficacy to guide clinical decision-making.
[0005] How to achieve drug sensitivity testing in vitro to guide personalized medicine has become an urgent technical problem to be solved. Utility Model Content
[0006] To address the problems existing in the prior art, this utility model provides a microfluidic chip, comprising:
[0007] A substrate, with a cover plate covering the top of the substrate, and multiple flow channels formed between the substrate and the cover plate, each flow channel having a drug delivery port at one end and a first solenoid valve at the other end;
[0008] Multiple sets of three-dimensional culture chambers are arranged sequentially between the first solenoid valve and the second solenoid valve along the extension direction of the corresponding flow channel. The flow channel protrudes in an arc shape towards the cover plate where it flows through each of the three-dimensional culture chambers.
[0009] Each of the three-dimensional culture chambers has an input conduit toward the first solenoid valve and an outlet toward the second solenoid valve, the input conduit and the outlet communicating with the flow channel, and the diameter of the input conduit being smaller than the diameter of the outlet.
[0010] Preferably, the administration port is connected to one end of each of the flow channels via an administration conduit.
[0011] Preferably, each group of three-dimensional culture chambers includes multiple three-dimensional culture chambers, and all three-dimensional culture chambers are arranged in an array on each flow channel.
[0012] Preferably, it also includes a flushing pipe, which is sleeved on the input pipe. One end of the flushing pipe is fixed to the outer wall of the input pipe, and the other end is connected to the corresponding three-dimensional culture chamber. The inner diameter of the cross-section of the flushing pipe gradually increases along the direction toward the three-dimensional culture chamber.
[0013] The side of the input pipe is connected to the flushing pipe and is connected via a two-way valve.
[0014] Preferably, the two-way valve is a flow valve.
[0015] Preferably, a biosensor is provided at the end of each flow channel located away from the three-dimensional culture chamber from the second solenoid valve.
[0016] Preferably, the first solenoid valve and the second solenoid valve are air pump valves.
[0017] The above technical solution has the following advantages or beneficial effects:
[0018] 1) By providing multiple flow channels through microfluidic chips, multiple drug treatment regimens and combination strategies can be evaluated simultaneously, enabling high-throughput continuous multi-day culture of micro-tumors. This can be used in fields such as clinical personalized medication regimen development, new drug research and development, and basic medical research, thereby promoting precision treatment of lung cancer.
[0019] 2) Each channel provides multiple three-dimensional culture chambers, enabling each drug regimen to act on multiple three-dimensional culture chambers simultaneously, while also reflecting the heterogeneity of different three-dimensional tumor cells from the same patient. Furthermore, the three-dimensional culture chambers within the same channel are interconnected, making it closer to the real human environment.
[0020] 3) The flow channel protrudes in an arc shape towards the cover plate at the point where it flows through each three-dimensional culture chamber, so that multiple three-dimensional culture chambers can be drugged at the same time without affecting each other.
[0021] 4) The inlet channel has a relatively small diameter, which allows for more precise control of the drug dosage, while the outlet of the three-dimensional culture chamber has a relatively large diameter, which allows the three-dimensional tumor spheres to pass through smoothly, reducing the difficulty of chip washing.
[0022] 5) By designing the flushing pipe with a gradually increasing inner diameter of the cross-section, the flushing solution can be buffered and depressurized before entering the three-dimensional culture chamber, preventing excessive flushing pressure from damaging the cells of the three-dimensional tumor spheres and thus affecting the subsequent detection of cytokines.
[0023] 6) By setting a biosensor at the end of the flow channel, it is possible to detect cytokines in three-dimensional tumor spheres after drug sensitivity reaction, thereby reflecting the drug sensitivity reaction results of the corresponding drugs. Attached Figure Description
[0024] Figure 1 A schematic diagram of the structure of a microfluidic chip is shown in a preferred embodiment of this utility model.
[0025] Figure 2 This is a schematic diagram of the structure of the three-dimensional culture chamber in a preferred embodiment of the present invention. Detailed Implementation
[0026] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments. The present invention is not limited to this embodiment; other embodiments that conform to the spirit of the present invention may also fall within its scope.
[0027] In a preferred embodiment of this utility model, based on the above-mentioned problems existing in the prior art, a microfluidic chip is now provided, such as... Figure 1 and Figure 2 As shown, it includes:
[0028] The substrate 1 has a cover plate 2 covering the top of the substrate 1, and multiple flow channels 3 are formed between the substrate 1 and the cover plate 2. One end of each flow channel 3 is connected to the administration port 4 and is provided with a first solenoid valve 5, and the other end is provided with a second solenoid valve 6.
[0029] Multiple sets of three-dimensional culture chambers 7 are arranged sequentially between the first solenoid valve 5 and the second solenoid valve 6 along the extension direction of the corresponding flow channel 3. The flow channel 3 protrudes in an arc shape towards the cover plate 2 where it flows through each three-dimensional culture chamber 7.
[0030] Each three-dimensional culture chamber 7 has an input pipe 71 facing the first solenoid valve 5 and an outlet 72 facing the second solenoid valve 6. The input pipe 71 and the outlet 72 are connected to the flow channel 3, and the diameter of the input pipe 71 is smaller than the diameter of the outlet 72.
[0031] Specifically, in this embodiment, the substrate 1 is preferably prepared using polydimethylsiloxane (PDMS) soft etching technology. Specifically, a template containing the aforementioned flow channels 2 and the three-dimensional culture chamber 7 can be prepared using 3D printing and soft photolithography, followed by casting of the PDMS prepolymer, and then thermal curing and sealing. The cover plate 2 is made of transparent material, facilitating observation of the morphological changes of the three-dimensional tumor spheres in the three-dimensional culture chamber 7 and drug administration.
[0032] In practical use, fresh tumor samples from patients can first undergo single-cell processing in vitro, followed by suspension culture in low-adhesion culture dishes using a microtumor-specific culture medium for 2-7 days. This allows primary tumor cells to rapidly proliferate and self-assemble with cells in the microenvironment to form three-dimensional tumor spheres with a diameter of 40-300 μm. These three-dimensional tumor spheres are then placed in three-dimensional culture chambers 7, preferably containing 30-50 spheres per chamber 7. Each flow channel 2 should have at least three three-dimensional culture chambers 7 for parallel control.
[0033] After the three-dimensional tumor spheres are placed, an injection pump can be connected to the administration port 4 via a tubing. The injection pump injects the drug to be tested for drug sensitivity into the administration port 4. The drug flows into the corresponding three-dimensional culture chambers 7 through the corresponding flow channels 3. Specifically, for each flow channel 3, when drug injection is required, the corresponding first solenoid valve 5 is first opened to allow the drug to enter the three-dimensional culture chamber 7, and the second solenoid valve 6 is closed to prevent the drug from being flushed out of the microfluidic chip. Then, the injection pump is controlled to inject the corresponding drug, realizing high-throughput continuous multi-day culture of microtumors on the microfluidic chip to observe the drug sensitivity test results of the cultured tumor cells.
[0034] Among them, such as Figure 1 As shown, the side of the substrate 1 can also be provided with connection holes for each of the first solenoid valves 5, which are electrically connected to each of the first solenoid valves 5. Each connection hole can be connected to the controller by an outward lead wire to realize the sequential control of each of the first solenoid valves 5.
[0035] Preferably, each flow channel 3 flows sequentially through each three-dimensional culture chamber 7. When the drug is injected, it is delivered to the first three-dimensional culture chamber 7 through the flow channel 3 at the front end and then splits into two paths. One path is input into the three-dimensional culture chamber 7 through the input pipe 71, and the other path bypasses the first three-dimensional culture chamber 7 and is delivered to the next three-dimensional culture chamber 7, and so on. There is no need to inject the first three-dimensional culture chamber 7 first and then inject the second three-dimensional culture chamber 7, which reduces the delay of drug injection in each three-dimensional culture chamber 7.
[0036] More preferably, the outlet diameter of each three-dimensional culture chamber 7 is the same as the diameter of the flow channel 3 and is larger than the diameter of the input pipe 71. This allows for more precise control of the drug dosage while ensuring that the three-dimensional tumor spheres can pass smoothly through the outlet, reducing the difficulty of chip rinsing after drug sensitivity culture.
[0037] In a preferred embodiment of the present invention, the administration port 4 is connected to one end of each flow channel 3 via the administration conduit 41.
[0038] In a preferred embodiment of the present invention, each group of three-dimensional culture chambers 7 includes multiple three-dimensional culture chambers 7, and all three-dimensional culture chambers 7 are arranged in an array on each flow channel 3.
[0039] Specifically, in this embodiment, by designing each three-dimensional culture cavity 7 to be distributed in an array, the three-dimensional culture cavities 7 located on different flow channels 3 can be aligned, which facilitates intuitive comparison of the sensitivity detection results of different drugs.
[0040] In a preferred embodiment of the present invention, a flushing pipe 8 is further included, which is sleeved on the input pipe 71. One end of the flushing pipe 8 is fixed to the outer wall of the input pipe 71, and the other end is connected to the corresponding three-dimensional culture chamber 7. The inner diameter of the cross section of the flushing pipe 8 gradually increases along the direction toward the three-dimensional culture chamber 7.
[0041] The side of the inlet pipe 71 is connected to the flushing pipe 8 and is connected via a two-way valve 9.
[0042] Specifically, due to the small diameter of the input pipe 71, if it were used as the flushing pipe 8 during rinsing, the small diameter would result in a high flow rate and pressure of the flushing fluid reaching the three-dimensional culture chamber 7, which could easily damage the three-dimensional tumor spheres after drug sensitivity testing. Therefore, in this embodiment, a flushing pipe 8 with a gradually increasing cross-sectional inner diameter is designed. When drug injection is required, the input pipe 71 and the flushing pipe 8 are isolated by a two-way valve 9, ensuring that the drug can only reach the corresponding three-dimensional culture chamber 7 through the input pipe 71 without affecting the drug injection volume. When the microfluidic chip needs to be flushed after drug sensitivity testing, the input pipe 71 and the flushing pipe 8 are connected by the two-way valve 9, and the original outlet of the input pipe 71 is isolated by the two-way valve 9. At this time, the flushing solution flows through the input pipe 71 to the two-way valve 9 and then into the flushing pipe 8. The gradually increasing cross-sectional inner diameter of the flushing pipe 8 effectively reduces the flow rate and pressure of the flushing solution, thus avoiding excessive mechanical stress or damage to the cells.
[0043] In a preferred embodiment of this utility model, the two-way valve 9 is a flow valve.
[0044] Specifically, in this embodiment, the two-way valve 9 is designed as a flow valve, which can close the input pipe 71 when the amount of drug injected reaches a preset threshold during drug injection, so that the corresponding three-dimensional culture chamber 7 no longer receives drugs, and all drugs are sent to the next three-dimensional culture chamber 7 through the flow channel 3 until all flow valves reach the preset threshold, then the injection pump is controlled to be turned off, and the drug injection process ends.
[0045] In a preferred embodiment of the present invention, a biosensor is provided at the end of each flow channel 3 located away from the three-dimensional culture chamber 7 of the second solenoid valve 6.
[0046] Specifically, in this embodiment, the biosensor can detect cytokines in the three-dimensional tumor spheres after drug sensitivity testing to determine the specific points where the drug acts. These cytokines include, but are not limited to, IFNβ, TNF, and IL-6.
[0047] IFNβ is a type of interferon (IFN), belonging to the low-molecular-weight proteins (mainly glycoproteins) with various biological activities. It possesses multiple biological activities, including antiviral, antitumor, and immunomodulatory effects. It does not directly kill or inhibit viruses, but rather inhibits viral replication by inducing cell surface receptors to produce antiviral proteins. In cancer treatment, IFNβ exerts its antitumor effects by activating immune cells, inhibiting tumor angiogenesis, and inducing tumor cell apoptosis. However, the efficacy of IFNβ and patient response may vary due to individual differences. By detecting IFNβ levels, the sensitivity of patients to cancer drugs can be indirectly assessed, thereby guiding the development of personalized treatment plans.
[0048] TNF is the abbreviation for tumor necrosis factor, a family of cytokines that cause necrosis of tumor cells. TNF can be divided into two types: TNF-α and TNF-β. TNF-α is mainly produced by activated monocytes / macrophages, while TNF-β is mainly produced by activated T cells. It has important biological functions such as antiviral, antitumor, immunomodulatory, inflammatory response promotion, pyrogenicity, and cachexia induction, and can be used in tumor treatment. In particular, TNF-α plays a crucial role in tumor treatment. It can induce tumor cell apoptosis, enhance the killing activity of immune cells, and promote inflammatory responses, thereby exerting an antitumor effect. The detection of TNF is of great significance in tumor drug sensitivity testing for the following reasons:
[0049] Predicting therapeutic efficacy: TNF levels can reflect the sensitivity of tumor cells and the activity of immune cells, thus predicting the efficacy of tumor drugs.
[0050] Monitoring changes in the condition: During treatment, changes in TNF levels can reflect changes in the condition and the effectiveness of treatment, which helps to adjust the treatment plan in a timely manner.
[0051] IL-6 is a type of interleukin (IL), a pleiotropic cytokine produced by various cells in the body, such as activated T cells, B cells, monocytes, and endothelial cells. It participates in a variety of physiological and pathophysiological processes, playing a particularly important role in inflammation and immune responses. In cancer treatment, changes in IL-6 levels can reflect the proliferation, invasion, and metastasis of tumor cells, as well as the activity of immune cells. Therefore, IL-6 detection has the following significance in tumor drug sensitivity testing:
[0052] Assessing tumor activity: IL-6 levels can reflect the activity of tumor cells, thereby assessing the malignancy and prognosis of the tumor.
[0053] Predicting drug efficacy: Changes in IL-6 levels can predict the killing effect of tumor drugs on tumor cells, thereby guiding the formulation of treatment plans.
[0054] Monitoring treatment response: Changes in IL-6 levels during treatment can reflect treatment response and changes in the condition, helping to adjust the treatment plan in a timely manner.
[0055] The detection of cytokines IFNβ, TNF, and IL-6 not only has independent predictive and monitoring value in tumor drug susceptibility testing, but their interactions and balance also have significant clinical implications. For example, IFNβ and TNF can synergistically exert anti-tumor effects, while elevated IL-6 may indicate drug resistance and immune evasion mechanisms in tumor cells. Therefore, comprehensive detection of these cytokine levels can provide a more complete assessment of the efficacy of tumor drugs and patient response, offering a more accurate basis for developing personalized treatment plans.
[0056] The type and specific model of the aforementioned biosensor (not shown in the figure) are not limited, as long as it can detect the aforementioned cytokines.
[0057] In a preferred embodiment of this utility model, the first solenoid valve 5 and the second solenoid valve 6 are air pump valves.
[0058] In summary, the microfluidic chip of this invention can highly simulate the immune microenvironment characteristics of lung cancer, and can simultaneously evaluate multiple drug treatment regimens and combination strategies. It can be used in fields such as clinical personalized medication regimen formulation, new drug development and basic medical research, thereby promoting precision treatment of lung cancer.
[0059] The above description is only a preferred embodiment of the present utility model and does not limit the implementation method and protection scope of the present utility model. Those skilled in the art should realize that all solutions obtained by equivalent substitutions and obvious changes made using the content of this specification and illustrations should be included within the protection scope of the present utility model.
Claims
1. A microfluidic chip, characterized in that, include: A substrate, with a cover plate covering the top of the substrate, and multiple flow channels formed between the substrate and the cover plate, each flow channel having a drug delivery port at one end and a first solenoid valve at the other end; Multiple sets of three-dimensional culture chambers are arranged sequentially between the first solenoid valve and the second solenoid valve along the extension direction of the corresponding flow channel. The flow channel protrudes in an arc shape towards the cover plate where it flows through each of the three-dimensional culture chambers. Each of the three-dimensional culture chambers has an input pipe toward the first solenoid valve and an outlet toward the second solenoid valve, the input pipe and the outlet communicating with the flow channel, and the diameter of the input pipe being smaller than the diameter of the outlet; It also includes a flushing pipe, which is sleeved on the input pipe. One end of the flushing pipe is fixed to the outer wall of the input pipe, and the other end is connected to the corresponding three-dimensional culture chamber. The inner diameter of the cross-section of the flushing pipe gradually increases along the direction toward the three-dimensional culture chamber. The side of the input pipe is connected to the flushing pipe and is connected via a two-way valve.
2. The microfluidic chip according to claim 1, characterized in that, The administration port is connected to one end of each of the flow channels via an administration conduit.
3. The microfluidic chip according to claim 1, characterized in that, Each group of three-dimensional culture chambers contains multiple three-dimensional culture chambers, and all three-dimensional culture chambers are arranged in an array on each flow channel.
4. The microfluidic chip according to claim 1, characterized in that, The two-way valve is a flow valve.
5. The microfluidic chip according to claim 1, characterized in that, Each of the flow channels has a biosensor positioned at the end opposite to the three-dimensional culture chamber of the second solenoid valve.
6. The microfluidic chip according to claim 1, characterized in that, The first solenoid valve and the second solenoid valve are air pump valves.