Organ-on-a-chip

Abstract

The application discloses an organ chip. The organ chip comprises a distribution layer, a porous membrane layer, a culture layer and a sensing layer which are sequentially stacked, the distribution layer is provided with a distribution flow channel, the distribution flow channel forms a feeding port on the distribution layer, the porous membrane layer is used for cell growth and is provided with micropores, the culture layer is provided with a culture unit and a communication hole, the culture unit is used for culturing organs or tissues, the micropores are communicated with the feeding port and the culture unit, and the sensing layer comprises a sensing electrode, and the communication hole is communicated with the culture unit and the sensing electrode. The porous membrane layer and the culture layer can culture cells and organs, realize construction of an organ model, the distribution layer can inject drugs, the sensing layer can monitor organ parameters in real time, and one-stop integration of culture, injection and detection functions is realized.

Description

Technical Field

[0001] This invention relates to the field of microfluidic chip technology, and in particular to an organ-on-a-chip. Background Technology

[0002] In drug development, preclinical evaluation relies on in vitro cell models and animal models. However, traditional two-dimensional cell culture cannot simulate the three-dimensional structure, intercellular interactions, and dynamic microenvironment in vivo. Animal models, on the other hand, suffer from problems such as large species differences, high costs, long cycles, and ethical controversies, resulting in approximately 90% of drugs that pass preclinical evaluation failing in human trials. Therefore, there is an urgent need for in vitro models that can more accurately predict human responses.

[0003] To address this, organ-on-a-chip technology has emerged. It simulates the structural and functional units of organs by constructing physiological microsystems on microfluidic chips. For example, research has developed chips for the culture and drug testing of single organs or organoids such as the lungs, liver, intestines, and blood-brain barrier. However, most existing chips have relatively limited functionality, separating culture, drug administration, and detection processes, making it difficult to achieve a complete closed loop—from biomimetic culture and controlled drug exposure to real-time, multi-parameter efficacy evaluation—on a compact, standardized platform. Summary of the Invention

[0004] This invention aims to address at least one of the technical problems existing in the prior art. To this end, this invention proposes an organ-on-a-chip that integrates culture, injection, and detection functions.

[0005] An organ-on-a-chip according to a first aspect of the present invention includes a dispensing layer, a porous membrane layer, a culture layer, and a sensing layer stacked sequentially. The dispensing layer is provided with dispensing channels, and the dispensing channels form a feed port on the dispensing layer. The porous membrane layer is used for cell growth and is provided with micropores. The culture layer is provided with culture units and connecting holes. The culture units are used for culturing organs or tissues, and the micropores connect the feed port and the culture units. The sensing layer includes sensing electrodes, and the connecting holes connect the culture units and the sensing electrodes.

[0006] The organ-on-a-chip according to embodiments of the present invention has at least the following beneficial effects: the porous membrane layer and the culture layer can culture cells and organs to realize the construction of organ models; the distribution layer can inject drugs; and the sensing layer can monitor organ parameters in real time, realizing one-stop integration of culture, injection and detection functions.

[0007] According to some embodiments of the present invention, the organ-on-a-chip further includes a flow guiding layer, which is stacked on the side of the distribution layer opposite to the porous membrane layer. The flow guiding layer is provided with an inlet, an outlet, and a detection window. The inlet is connected to the distribution channel, and the detection window is connected to the culture unit. The culture layer is also provided with an outlet hole, which is connected to the culture unit and the outlet.

[0008] According to some embodiments of the present invention, the distribution channel includes a hierarchical channel, the hierarchical channel including at least one branching unit, the branching unit including a first branch, a second branch and a third branch, the second branch and the third branch being connected to the same end of the first branch.

[0009] According to some embodiments of the present invention, the organ-on-a-chip further includes a damper, and the damper is provided at least one of the dispensing channel, the porous membrane layer, the culture layer and the sensing layer.

[0010] According to some embodiments of the present invention, the organ-on-a-chip further includes a mixer, wherein the mixer is disposed at least at one location of the dispensing channel and the sensing layer.

[0011] According to some embodiments of the present invention, the culture unit includes a first unit and a second unit, wherein the volume of the first unit is larger than the volume of the second unit.

[0012] According to some embodiments of the present invention, a plurality of second units are arranged around the first unit, wherein the first unit is used for culturing organs and the second units are used for culturing cells.

[0013] According to some embodiments of the present invention, the sensing layer includes an expansion interface that communicates with the communication hole and is used to connect a sensor.

[0014] According to some embodiments of the present invention, the organ-on-a-chip further includes a physical stimulation module, which is capable of applying at least one of shear force, pressure, deformation force, extrusion force, and electrical stimulation.

[0015] According to some embodiments of the present invention, the porous membrane layer is detachably connected to the distribution layer and the culture layer, and the porous membrane layer includes a first porous membrane layer and a second porous membrane layer that can be selectively used; wherein the size of the micropores in the first porous membrane layer is smaller than the size of the micropores in the second porous membrane layer; and / or, the material of the first porous membrane layer is different from the material of the second porous membrane layer.

[0016] Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Attached Figure Description

[0017] The present invention will be further described below with reference to the accompanying drawings and embodiments, wherein: Figure 1 This is a schematic diagram of an organ-on-a-chip according to an embodiment of the present invention; Figure 2 A schematic diagram of the flow distribution channels; Figure 3 A schematic diagram of a mixer; Figure 4 This is a schematic diagram of the culture layer.

[0018] Figure label: 100, 110, 120, 130, 140, 140; Distribution layer 200, distribution channel 210, feed port 211, grading channel 220, damper 230, mixer 240, first mixing 250, doxorubicin mother liquor 251, culture medium 252, second mixing 260, third mixing 270, third-stage solution 271, branching unit 280, first branch 281, second branch 282, third branch 283, fourth branch 284, serpentine channel 290; Porous membrane layer 300, micropores 310; Culture layer 400, culture unit 410, first unit 411, second unit 412, organoid sphere 420, discharge pore 430; Sensing layer 500, sensing electrode 510, expansion interface 520. Detailed Implementation

[0019] Embodiments of the present invention are described in detail below. Examples of these embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. 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.

[0020] In the description of this invention, it should be understood that the orientation descriptions, such as up, down, front, back, left, right, etc., are based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limiting this invention.

[0021] In the description of this invention, "several" means one or more, "multiple" means two or more, "greater than," "less than," and "exceeding" are understood to exclude the stated number, while "above," "below," and "within" are understood to include the stated number. The use of "first" and "second" in the description is merely for distinguishing technical features and should not be construed as indicating or implying relative importance, or implicitly indicating the number of indicated technical features, or implicitly indicating the order of the indicated technical features.

[0022] In the description of this invention, unless otherwise explicitly defined, terms such as "set up," "install," and "connect" should be interpreted broadly, and those skilled in the art can reasonably determine the specific meaning of the above terms in this invention in conjunction with the specific content of the technical solution.

[0023] In the description of this invention, the terms "one embodiment," "some embodiments," "illustrative embodiment," "example," "specific example," or "some examples," etc., refer to specific features, structures, materials, or characteristics described in connection with that embodiment or example, which are included in at least one embodiment or example of the invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0024] To address the problems in related technologies, this application integrates drug dispensing, organ culture layers, and real-time monitoring into a single, compact, and standardized culture and testing platform, thereby improving the effectiveness of preclinical evaluation.

[0025] The organ-on-a-chip embodiment of this application is described below with reference to the accompanying drawings. It should be noted that... Figure 1 The flow channels are simply illustrated with arrows; please refer to the diagram for details. Figure 2 Configure the flow distribution channels. Figure 3 The serpentine flow channel can be used as a mixer or as a damper.

[0026] Reference Figure 1According to a first aspect of the present invention, an organ-on-a-chip includes a distribution layer 200, a porous membrane layer 300, a culture layer 400, and a sensing layer 500 stacked sequentially. The distribution layer 200 is provided with a distribution channel 210, and a feed port 211 is formed on the distribution layer 200. The porous membrane layer 300 is used for cell growth and is provided with micropores 310. The culture layer 400 is provided with a culture unit 410 and a connecting hole (not shown in the figure). The culture unit 410 is used for culturing organs or tissues, and the micropores 310 connect the feed port 211 and the culture unit 410. The sensing layer 500 includes a sensing electrode 510, and the connecting hole connects the culture unit 410 and the sensing electrode 510. The porous membrane layer 300 and the culture layer 400 can culture cells and organs to build organ models, the distribution layer 200 can inject drugs, and the sensing layer 500 can monitor organ parameters in real time, realizing one-stop integration of culture, injection and detection functions.

[0027] The one-stop integration of various functions not only realizes the functions of each layer, but also plays a role in dynamic interaction and synergistic gain effect. Through precise flow channel interconnection and functional partitioning, this application realizes a closed-loop linkage mechanism of "fluid distribution-barrier simulation-real-time monitoring". The drug gradient generated by the distribution layer 200 permeates to the culture layer 400 through the porous membrane layer 300, simulating the in vivo drug transport across the barrier. The integrated sensing layer 500 monitors changes in transmembrane resistance or other values ​​in real time, and adjusts the perfusion rate of the distribution layer 200 accordingly to form an adaptive drug exposure environment. The three major functions of biomimetic microenvironment construction, dynamic drug exposure and gradient generation, and in-situ physiological parameter monitoring are organically integrated into a compact chip, overcoming the problems of cumbersome operation, sample damage and error introduction caused by the separation of culture, drug addition and detection links in traditional methods. It realizes a complete and controllable miniaturized experimental closed loop from cell culture to drug efficacy data acquisition.

[0028] Reference Figure 1 According to some embodiments of the present invention, the organ-on-a-chip further includes a flow guiding layer 100, which is stacked on the side of the distribution layer 200 opposite to the porous membrane layer 300. The flow guiding layer 100 is provided with an inlet 110, an outlet 120, and a detection window 130. The inlet 110 is connected to the distribution channel 210, and the detection window 130 is connected to the culture unit 410. The culture layer 400 is also provided with a discharge port 430, which is connected to the culture unit 410 and the outlet 120. As the outermost layer of the organ-on-a-chip, the flow guiding layer 100, while sealing the distribution channel 210, provides the inlet 110 for drug delivery to the distribution channel 210, provides the outlet 120 for draining the perfused waste liquid, and provides the detection window 130 for direct observation of the inner side of the organ-on-a-chip.

[0029] Specifically, multiple inlet ports 110, outlet ports 120, and detection windows 130 can be adaptively configured. Multiple inlet ports 110 can meet the perfusion requirements of different drugs or the requirements of drug concentration adjustment. For example, one inlet port 110 can serve as the main inlet for sample injection, while several inlet ports 110 can serve as sub-inlet ports, participating in the concentration mixing of the drug solution during the secondary distribution process in the distribution channel 210, ultimately forming multiple drug solutions of different concentrations. Multiple outlet ports 120 can respectively collect waste liquid discharged from different culture units 410. For example, one outlet port 120 can correspond to one culture unit 410, or one outlet port 120 can correspond to multiple culture units 410 within a region. Multiple detection windows 130 can observe different locations within the organ-on-a-chip, for example, observing the monolayer of cells formed on the porous membrane layer 300, the organs formed on the culture layer 400, and the cells formed on the culture layer 400.

[0030] Reference Figure 1 and Figure 2 According to some embodiments of the present invention, the distribution channel 210 includes a graded channel 220, which includes at least one branching unit 280. The branching unit 280 includes a first branch 281, a second branch 282, and a third branch 283, with the second branch 282 and the third branch 283 connected to the same end of the first branch 281. The branching unit 280 is capable of diverting the liquid medicine in the first branch 281 to achieve a single distribution of the liquid medicine.

[0031] Specific reference Figure 2 The distribution channel 210 can be equipped with multiple branching units 280 to distribute the medicine multiple times. Each branch that is distributed can be regarded as a first branch 281. For example, if the medicine distributed by the second branch 282 of the previous level is distributed again, the second branch 282 can be regarded as the first branch 281 of the current level. As needed, a fourth branch 284 can be set on the basis of the second branch 282 and the third branch 283 to distribute the medicine of the first branch 281 into three parts. More branches can also be set to distribute multiple parts.

[0032] Furthermore, the second branch 282 and the third branch 283 can be branches with the same flow capacity or branches with different flow capacities. For example, the flow capacity of the second branch 282 is twice that of the third branch 283, so that 2 / 3 of the medicine is distributed through the second branch 282 and 1 / 3 of the medicine is distributed through the third branch 283.

[0033] The hierarchical flow channel 220 can specifically form a tree-like flow distribution network, such as... Figure 2As shown, taking the preparation of doxorubicin as an example, the doxorubicin stock solution 251 and culture medium 252 are first mixed 250 times, then split to form two first-stage solutions. These two first-stage solutions are then split and mixed a second time 260 times to form four second-stage solutions. After being split and mixed a third time 270 times, the four second-stage solutions form twelve third-stage solutions. These twelve third-stage solutions are then transported in parallel to their respective culture units 410. The twelve third-stage solutions can be twelve solutions of different concentrations, or solutions with some of the same concentration. For example, eight solutions of different concentrations can be prepared, with three portions of each of two concentrations prepared. The specific preparation can be adapted to meet specific needs.

[0034] The microfluidic network designed in the distribution layer 200 can mix single or multiple drug solutions in a preset ratio to generate a series of concentration gradients, which are then distributed in parallel to the individual culture units 410 below. Combined with the mixer 240 and damper 230, the accuracy and stability of concentration generation are further ensured. Through active mixing or passive diffusion principles, a series of precise and stable concentration gradients are automatically generated and synchronously and independently distributed to the multiple culture units 410 arranged in an array below via parallel flow channels. This design allows a single chip to simultaneously perform dose-response relationship testing at multiple concentrations of a single drug, or to conduct parallel screening of multiple different drugs, greatly improving the throughput and efficiency of drug testing.

[0035] Reference Figure 2 According to some embodiments of the present invention, the organ-on-a-chip further includes a damper 230, which is provided at least at one location in the distribution channel 210, the porous membrane layer 300, the culture layer 400, and the sensing layer 500. The damper 230 can suppress flow pulsation and stabilize flow rate and pressure. Installed at the inlet front end of the distribution channel 210, it can smooth out periodic pulsations generated by the pump delivering the drug solution and provide a stable initial flow rate. Installed at the first branch 281 of the branching unit 280 of the distribution channel 210, it can ensure that the instantaneous flow rate and pressure of each branch are consistent at the distribution unit, thereby generating a precise concentration gradient or achieving uniform distribution. Installed at the inlet front end of the porous membrane layer 300, the inlet front end of the culture layer 400, and the inlet front end of the sensing layer 500, it can isolate pressure surges from other upstream parts (such as valve opening and closing, disturbances from other branches), creating a local stable flow field for sensitive areas.

[0036] Specifically, the damper 230 can be an active damper or a passive damper. It can be a standalone device or a damping structure formed within the flow channel. For example, a microfluidic vibration reduction chip based on electrorheological fluid can be used as an active standalone device damper 230. By controlling the electric field, the magnitude of the damping force can be adjusted in real time and precisely, achieving more intelligent control of vibration.

[0037] For example, a microfluidic gas damper integrated into the liquid inlet front end of the distribution channel 210 serves as a passive, single-section damper. This microfluidic gas damper employs a multi-layer structure, containing an elastic membrane. The membrane's deformation absorbs pressure fluctuations, achieving miniaturization and passive stabilization through a passive approach. The elastic membrane can be directly integrated into the flow channel; for instance, a cavity can be fabricated beside the flow channel and covered with an elastic membrane (such as PDMS). When the pressure increases, the membrane deforms outward, increasing the cavity volume and storing fluid; when the pressure decreases, the membrane rebounds, releasing the fluid and thus absorbing pulsations.

[0038] For example, designs in the flow channel can incorporate structures that abruptly contract or expand (such as flares or chambers) or gradually contracting / expanding nozzles / diffusers. When fluid flows through these areas, the dramatic changes in velocity lead to pressure loss and eddy formation, thus consuming energy. Alternatively, a very narrow channel can be created within the flow channel, or a porous medium can be embedded. According to the Hagen-Poiseuille law, flow resistance is inversely proportional to the fourth power of the hydraulic radius of the channel; a small change in cross-sectional area can significantly increase flow resistance and stabilize the flow rate. Or, refer to... Figure 3 By setting a portion of the flow channel as a serpentine flow channel 290, the flow path of the fluid is greatly increased and its direction is changed multiple times. The viscous friction between the fluid and the wall and between the fluid and its interior continuously consumes its energy, thereby stabilizing the flow.

[0039] According to some embodiments of the present invention, the organ-on-a-chip further includes a mixer 240, and the mixer 240 is disposed at least at one location in the distribution channel 210 and the sensing layer 500. The mixer 240 can thoroughly and rapidly mix different fluids uniformly. Disposing the mixer 240 at the inlet end of the distribution channel 210 enables rapid and uniform mixing of the drug solution. In some cases requiring optical detection (such as photometric absorption), placing the mixer 240 at the inlet end of the sensing layer 500 ensures that the liquid flowing through the detection window 130 is uniform, avoiding fluctuations in the detection signal due to uneven mixing, thereby ensuring data accuracy.

[0040] Specifically, the mixer 240 can be selected as an active mixer, such as an ultrasonic micro mixer, a magnetic stirring micro mixer, an electric active micro mixer, a pressure perturbation mixer, etc., which uses ultrasound, magnetism, electricity or pressure to achieve active stirring and achieve rapid and efficient mixing.

[0041] Mixer 240 can also be selected as a passive mixer (directly integrated into the flow channel), for example Figure 3Vortex mixers utilize curved flow channels (such as serpentine channels 290) to generate secondary flows like Dean vortices during fluid flow, thereby stretching, folding, and reorganizing fluids from different flow layers to achieve convective mixing. For example, laminar flow mixers first separate two fluid streams into multiple thin streams, then arrange them in alternating layers, greatly increasing the contact area and thus achieving rapid mixing through diffusion. Chaotic convective mixers, for instance, design specific asymmetric barriers, grooves, or contraction-expansion structures within the flow channel to generate chaotic streamlines as the fluid flows through, greatly promoting mixing. The flow channel can also be designed with a corrugated shape to achieve mixing.

[0042] According to some embodiments of the present invention, the culture unit 410 can culture cells, organoids, or tissue blocks for different conditions. The culture unit 410 forms microchannels through connecting holes at its bottom, communicating with the sensing layer 500 to achieve real-time detection. (Refer to...) Figure 1 and Figure 4 In some embodiments, the culture unit 410 includes a first unit 411 and a second unit 412, wherein the volume of the first unit 411 is larger than the volume of the second unit 412. The larger volume of the first unit 411 is used, for example, to form a culture chamber for culturing pre-fabricated organoids or tissue blocks. The smaller volume of the second unit 412 is used, for example, to form micropits for capturing cells to form organoid spheres 420.

[0043] Reference Figure 1 and Figure 4 Furthermore, multiple second units 412 are arranged around the first unit 411, where the first unit 411 is used for culturing organs and the second units 412 are used for culturing cells. Specifically, the first unit 411 serves as the central chamber for culturing tumor organoids, and vascular endothelial cells are cultured around the first unit 411 to construct a vascularized tumor model. Other target models can also be constructed by replacing the organoids in the first unit 411. The number and specific location of the second units 412 can be adaptively varied according to the target model.

[0044] On the culture layer 400, the culture units 410 can be independent or interconnected and can be arranged in an array, such as a 12×8 array, forming 96 circular micro-culture chambers. This array design makes organ-on-a-chip easily integrated with automated pipetting workstations and high-speed imaging systems, suitable for large-scale drug screening and personalized medicine. Combined with the dispensing channel 210, it allows for simultaneous testing of multiple drugs or different concentrations, achieving high-throughput parallel culture and delivery. Furthermore, the sensing layer 500 monitors the cells in the culture layer 400, enabling non-invasive, real-time monitoring of key physiological parameters during drug action (such as barrier integrity, metabolites, and cell viability), achieving in-situ real-time evaluation. Based on real-time data, it can react rapidly, achieving dynamic and controllable drug perfusion.

[0045] The porous membrane layer 300 serves as the boundary. The distribution channels 210 on top of the porous membrane layer 300 simulate the lumen of a blood vessel. The porous membrane layer 300 is made of biocompatible materials (such as polycarbonate and PDMS) and simulates physiological barriers (such as the blood-brain barrier and the intestinal epithelial barrier). The culture layer 400 below the porous membrane layer 300 simulates the lumen of a tissue, thus simulating the complex organ microenvironment, including the tissue-blood vessel interface, to stably culture three-dimensional organoids or co-culture multiple cells, achieving a high degree of biomimicry.

[0046] Specifically, the sensing electrode 510 is a miniaturized electrode pair, formed as a gold thin-film ring electrode through photolithography and deposition. The electrode leads are connected to the pads at the edge of the organ-on-a-chip, and the electrode positions are directly aligned with the culture units 410 above. The electrodes can be used to measure the transmembrane resistance of the cell layer in real time and non-invasively, quantitatively assessing the effect of drugs on barrier integrity. Compared with evaluating drug efficacy through endpoint methods (such as fluorescence staining), it is more effective to reflect the efficacy or toxic mechanism of drugs earlier and more sensitively by monitoring dynamic parameters such as transmembrane resistance in real time. For example, by monitoring the change in TEER value in transmembrane resistance, the effect of drugs on the integrity of tissue barriers such as vascular endothelium, intestinal epithelium, and blood-brain barrier (i.e., potential toxicity) can be quantitatively and sensitively assessed, or the functional state of the cell layer can be dynamically reflected. This provides a dynamic pharmacological and toxicological data dimension that cannot be obtained by traditional endpoint staining methods, which is conducive to rapid experimental adjustments.

[0047] Reference Figure 1 According to some embodiments of the present invention, the sensing layer 500 includes an expansion interface 520, which is connected to a connecting hole and is used to connect a sensor. The expansion interface 520 can integrate or subsequently connect other sensors, such as a pH sensor, an oxygen sensor, or a metabolite detection electrode, to provide richer monitoring data.

[0048] The following uses a blood-brain barrier model as an example to illustrate the model construction and drug testing in some embodiments. Human brain microvascular endothelial cells were seeded on the upper surface of the porous membrane layer 300, and astrocytes were seeded in the lower chamber (culture layer 400). Dynamic perfusion culture was performed for 5-7 days to form a mature barrier. The transmembrane resistance was continuously monitored using the sensing electrode 510 of the sensing layer 500. After stabilization, culture medium containing different concentrations of the tested nanomedicine (simulating the blood side) was perfused from the distribution layer 200. Changes in transmembrane resistance were monitored in real time to assess drug toxicity, and samples were taken from the culture layer 400 at specific time points. Drug concentration was detected by HPLC-MS, and its penetration rate was calculated.

[0049] The following uses a tumor pharmacodynamic model as an example to illustrate the model construction and drug testing in some embodiments. Patient-derived glioblastoma organoids are pre-cultured in culture unit 410 of culture layer 400. Different concentrations of anticancer drugs or the aforementioned barrier-crossing nanomedicines are perfused through dispensing channels. The possible acidification of the organoid microenvironment (indirectly reflecting metabolism) is monitored through sensing layer 500, and finally, live-cell fluorescence staining (such as Calcein-AM / PI) is performed through the top detection window 130. High-throughput imaging analysis is then used to analyze the drug's killing effect on the organoids.

[0050] In specific data analysis, combining real-time acquired transmembrane resistance data, drug penetration rate data, and endpoint efficacy data allows for a comprehensive evaluation of a nanomedicine delivery system's safety across the blood-brain barrier, penetration efficiency, and tumor-killing effect. The organ-on-a-chip technology described in this application provides a powerful platform for the synergistic evaluation of complex drug formulations.

[0051] According to some embodiments of the present invention, the organ-on-a-chip further includes a physical stimulation module 140 to further simulate the in vivo environment. The physical stimulation module 140 is capable of applying at least one of shear force, pressure, deformation force, extrusion force, and electrical stimulation.

[0052] For example, flexible films and pneumatic channels can be integrated into the flow-guiding layer 100 or other layers. Periodic vacuuming can deform the porous membrane layer 300, thereby applying mechanical stretching stimulation to the cells growing on the membrane, simulating respiration or peristalsis. For example, culture medium can be driven to flow within the microchannels using injection pumps, peristaltic pumps, etc., generating tangential frictional force on adherent cells. For example, uniform hydrostatic pressure or periodic pressure can be applied to cells or three-dimensional cultures within a closed microchamber using pneumatic or hydraulic systems. For example, the elastic basement membrane of cell growth can be stretched isometrically or uniaxially using pneumatic, magnetic, or mechanical devices. For example, micron / nanometer-scale "fishbone" or other geometric patterns can be fabricated on the cell growth substrate, stimulating cells through mechanical compression or contact guidance effects. For example, electrodes can be integrated on both sides of the microchannel to apply a controllable electric field for studying electroactive cells (such as cardiomyocytes and nerve cells) or electroporation transfection.

[0053] According to some embodiments of the present invention, the porous membrane layer 300 is detachably connected to the distribution layer 200 and the culture layer 400 to achieve modular replacement of the porous membrane layer 300. Specifically, the porous membrane layer 300 includes a first porous membrane layer 300 and a second porous membrane layer 300, which can be selectively used. In some embodiments, the size of the micropores 310 of the first porous membrane layer 300 is smaller than the size of the micropores 310 of the second porous membrane layer 300. In some embodiments, the materials of the first porous membrane layer 300 and the second porous membrane layer 300 are different. In some embodiments, the first porous membrane layer 300 and the second porous membrane layer 300 differ in both the size of the micropores 310 and their own materials. By replacing different porous membrane layers 300, different cells can be cultured. In addition, by adjusting the culture units 410 in the culture layer 400, different models can be quickly adapted and constructed, such as the blood-brain barrier model (evaluating drug penetration), the liver sinusoids model (evaluating metabolism and hepatotoxicity), the vascularized tumor model (evaluating the efficacy and metastasis of anticancer drugs), and the intestinal model (evaluating the absorption of oral drugs). This enables the creation of a reconfigurable biomimetic microenvironment for evaluating different properties of drugs, such as penetration, metabolism, and targeting efficacy, and has a wide range of applications.

[0054] The embodiments of the present invention have been described in detail above with reference to the accompanying drawings. However, the present invention is not limited to the above embodiments, and various changes can be made within the scope of knowledge possessed by those skilled in the art without departing from the spirit of the present invention. Furthermore, the embodiments of the present invention and the features thereof can be combined with each other unless otherwise specified.

Claims

1. An organ-on-a-chip, characterized in that, The device comprises a distribution layer, a porous membrane layer, a culture layer, and a sensing layer stacked sequentially. The distribution layer is provided with a distribution channel, which forms a feed port on the distribution layer. The porous membrane layer is used for cell growth and is provided with micropores. The culture layer is provided with culture units and connecting holes. The culture units are used for culturing organs or tissues, and the micropores connect the feed port and the culture units. The sensing layer includes a sensing electrode, and the connecting holes connect the culture units and the sensing electrodes.

2. The organ-on-a-chip according to claim 1, characterized in that, It also includes a flow guiding layer, which is stacked on the side of the distribution layer opposite to the porous membrane layer. The flow guiding layer is provided with an inlet, an outlet and a detection window. The inlet is connected to the distribution channel and the detection window is connected to the culture unit. The culture layer is also provided with a discharge hole, which is connected to the culture unit and the outlet.

3. The organ-on-a-chip according to claim 1, characterized in that, The distribution channel includes a hierarchical channel, which includes at least one branching unit. The branching unit includes a first branch, a second branch, and a third branch, with the second branch and the third branch connected to the same end of the first branch.

4. The organ-on-a-chip according to claim 1, characterized in that, It also includes a damper, and the damper is provided at least one of the distribution channel, the porous membrane layer, the culture layer and the sensing layer.

5. The organ-on-a-chip according to claim 1, characterized in that, It also includes a mixer, which is provided at least at one location of the distribution channel and the sensing layer.

6. The organ-on-a-chip according to claim 1, characterized in that, The culture unit includes a first unit and a second unit, wherein the volume of the first unit is larger than the volume of the second unit.

7. The organ-on-a-chip according to claim 6, characterized in that, Multiple second units are arranged around the first unit, the first unit being used for culturing organs and the second units being used for culturing cells.

8. The organ-on-a-chip according to claim 1, characterized in that, The sensing layer includes an expansion interface that connects to the connecting hole and is used to connect a sensor.

9. The organ-on-a-chip according to claim 1, characterized in that, It also includes a physical stimulation module, which is capable of applying at least one of shear force, pressure, deformation force, extrusion force, and electrical stimulation.

10. The organ-on-a-chip according to claim 1, characterized in that, The porous membrane layer is detachably connected to the distribution layer and the culture layer, and the porous membrane layer includes a first porous membrane layer and a second porous membrane layer that can be selectively used. Wherein, the size of the micropores in the first porous membrane layer is smaller than the size of the micropores in the second porous membrane layer; and / or, the material of the first porous membrane layer is different from the material of the second porous membrane layer.

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