Apparatus for constructing an open-vascularized organ chip, split open-vascularized organ chip and method of constructing the same

Abstract

The application discloses a device for constructing an open vascularized organ chip, comprising a substrate layer, a channel layer and a culture medium cavity layer; the channel layer has at least one channel unit; the channel unit has a first channel and two or more second channels, the second channels are provided with culture medium inlets and outlets, and the second channels have a preset height difference with the first channel; the culture medium cavity layer has a plurality of culture medium cavities; the substrate layer, the channel layer and the culture medium cavity layer are designed in a split mode, and when stacked from bottom to top, the culture medium cavities correspond to and communicate with the culture medium inlets and outlets one by one. Further, the application also discloses a method for constructing an open vascularized organ chip by using the device and an obtained open vascularized organ chip. The culture medium cavity layer and the channel layer of the organ chip are designed in a split mode, and based on this, high flow rate gap flow through the interior of the matrix gel and pipeline flow along the side wall of the matrix gel are realized in the chip.

Description

Technical Field

[0001] This invention belongs to the field of vascularized tissue construction technology, and specifically relates to an open vascularized organ-on-a-chip construction technology. Background Technology

[0002] There are many methods for constructing vascularized tissues on organ-on-a-chip platforms. Among them, the technique based on endothelial cell self-assembly has attracted widespread attention. This method specifically involves constructing multiple adjacent channels on a microfluidic chip, injecting a matrix gel containing endothelial cells and stromal cells into one of these channels. After the matrix gel solidifies, the endothelial cells spontaneously assemble within it to form an interconnected vascular network. Simultaneously, during vascular development, the chip generates interstitial flow through the matrix gel and channel flow along the matrix gel sidewalls, thereby directionally regulating vascular development. The morphology, structure, and function of the resulting vascular network are similar to those of capillaries in vivo, showing broad application prospects in in vitro model construction, drug screening, and regenerative medicine.

[0003] Currently, the construction of self-assembled vascular networks often relies on closed organ-on-a-chip (OIA). Closed OIA refers to a chip where all channels have sidewall structures and are connected to the outside world through input / output ports at both ends. Due to its closed channel structure, closed OIA allows for complex multi-channel designs and fluid actuation, easily achieving interstitial flow through the matrix gel and channel flow along the matrix gel sidewalls. However, closed OIA suffers from problems such as complex operation, nutrient deficiency near the injection port area, air bubble blockage, matrix gel leakage, and low throughput. To address these issues, open-vascularized OIA has been developed. Unlike closed OIA, open-vascularized OIA channels lack sidewall structures, thus eliminating the reliance on channel sidewalls for matrix gel confinement. Instead, it utilizes capillary flow of liquid within the chip structure, which has height differences, to confine the matrix gel within channels open on both sides. Due to the open channel side structure, open-vascularized OIA offers many advantages: sufficient exchange of nutrients between the matrix gel and the outside, low risk of air bubble blockage, and the ability to achieve higher throughput, among others. However, in open vascularized organ-on-a-chip systems, the sample loading process requires direct pipetting onto the bottom side of the open channel using a pipette. Excessive height of the chamber increases operational difficulty, so the height of the culture medium chamber often needs to be controlled within 1 cm. This limited height results in a low hydrostatic pressure difference across the matrix gel, making it difficult to easily apply high-velocity interstitial flow stimulation to the cells within the matrix gel using this pressure difference. Furthermore, due to the side-loading operation of the bottom matrix gel channel, the side of the matrix gel channel is often just a single culture medium chamber, making it difficult to create additional fluid channels and thus hindering the generation of channel flow on the matrix gel side. In addition, since open vascularized organ-on-a-chip systems are often fabricated using methods such as 3D printing and injection molding, their manufacturing costs are relatively high.

[0004] In summary, although open vascularized tissue microarrays have many advantages, they still suffer from drawbacks such as high manufacturing costs, inability to achieve high-velocity interstitial flow through the matrix gel, and inability to achieve channel flow along the matrix gel sidewalls. These drawbacks limit their application. Summary of the Invention

[0005] To address the aforementioned problems, this invention provides an apparatus for constructing open vascularized organ-on-a-chip, a split open vascularized organ-on-a-chip, and a method for constructing the same. The invention separates the culture medium chamber layer and the channel layer of a traditional open vascularized organ-on-a-chip. In use, a matrix gel is first added to the open matrix gel channels within the channel layer. Then, pressure-sensitive double-sided adhesive is used to bond the channel layer and the culture medium chamber layer together. Finally, culture medium is added to the culture medium chamber of the culture medium chamber layer. This enables high-velocity interstitial flow through the matrix gel interior and channel flow along the matrix gel sidewalls on the chip.

[0006] The first aspect of this invention discloses an apparatus for constructing an open vascularized organ-on-a-chip, comprising: a base layer; a channel layer having at least one channel unit; the channel unit having a first channel open on one side and two or more second channels connected to the side of the first channel, the second channels having two or more culture medium inlets and outlets, and a preset height difference between the second channels and the first channels; and a culture medium chamber layer having multiple culture medium chambers; the base layer, channel layer, and culture medium chamber layer are designed separately, and when stacked from bottom to top, the culture medium chambers of the culture medium chamber layer correspond one-to-one with and are connected to the culture medium inlets and outlets of the second channels of the channel layer.

[0007] As an optional solution, the channel layer is formed by heat-sealing a substrate and a cover sheet; the substrate and the cover sheet are respectively provided with hollow structures, which are used to form a first channel and a second channel; the height of the first channel is determined by the thickness of the substrate, and the height of the second channel is determined by the sum of the thicknesses of the substrate and the cover sheet; the thicknesses of the substrate and the cover sheet are not required to be the same.

[0008] As an optional solution, the thickness of both the substrate and the cover plate is in the range of 100-500μm, including the endpoint values; the height of the first channel is less than the height of the second channel, and the preset height difference is 100-500μm.

[0009] As an alternative, the channel unit has two second channels, which are respectively arranged on the side of the first channel; the second channel is provided with two culture medium inlets and outlets, which are respectively arranged at the ends of the second channel away from the first channel.

[0010] As an optional embodiment, the number of culture medium chambers in the culture medium chamber layer is consistent with the number of culture medium inlets and outlets in the channel layer; the height of the culture medium chamber is 1-5cm; the cross-section of the culture medium chamber is circular; the culture medium inlet and outlet are circular holes, and the diameter of the circular holes is the same as the diameter of the circular cross-section of the culture medium chamber.

[0011] As an alternative, the channel layer is made of acrylic or polycarbonate; the culture medium chamber layer is made of acrylic / polycarbonate; the base layer is made of a biocompatible material with a visible light transmittance greater than 90%; preferably glass or quartz.

[0012] As an alternative, when the device is used, the base layer, channel layer, and culture medium chamber layer are bonded together and fixed with pressure-sensitive double-sided adhesive.

[0013] A second aspect of this invention discloses a method for constructing an open vascularized organ-on-a-chip, and an apparatus for constructing an open vascularized organ-on-a-chip based on this invention and any of its alternative embodiments, comprising:

[0014] The base layer and the channel layer are fixedly connected, and the first channel is closed from top to bottom after the connection.

[0015] A matrix adhesive solution is injected into the first channel, and the matrix adhesive is confined within the first channel;

[0016] After the matrix adhesive has cured, the culture medium chamber layer and the channel layer are fixedly connected, and the culture medium chamber and the culture medium inlet and outlet of the second channel are connected one by one.

[0017] Culture medium is added to the culture medium chamber, and a hydrostatic pressure difference is generated between the sides of the matrix gel, which promotes interstitial flow within the matrix gel.

[0018] Optionally, the construction method further includes: creating a liquid level difference between two or more culture medium chambers connected by each second channel through a swinging operation, so as to form a pipe flow along the sidewall of the matrix gel within the second channel.

[0019] The third aspect of this invention discloses an open vascularized organ-on-a-chip, characterized in that it is constructed by the construction method of the open vascularized organ-on-a-chip described in the second aspect of this invention and any optional embodiment thereof.

[0020] The present invention has at least the following beneficial effects:

[0021] (1) The present invention adopts an open matrix gel channel design (the matrix gel channel has no sidewalls and its side is directly connected to the outside world), which overcomes the problems of complex operation, lack of nutrition in the near injection port area, air bubble blockage, matrix gel leakage, and low throughput of traditional closed vascularized organ-on-a-chip.

[0022] (2) The present invention adopts a separate design of channel layer and culture medium chamber layer. The culture medium chamber can be designed with any height according to the needs, thereby overcoming the problems that traditional open vascularized organ-on-a-chip cannot generate high-velocity gap flow through the matrix gel due to the limited height of the culture chamber, and cannot generate pipe flow along the side wall of the matrix gel because the side of the matrix gel channel is a single culture medium chamber.

[0023] (3) The channel layer designed in this invention can be prepared by hot pressing and sealing two or more layers of laser-cut acrylic sheets. Compared with 3D printing, injection molding, CNC machining and other processes, it can greatly reduce manufacturing costs while ensuring efficiency.

[0024] (4) The split open vascularized organ-on-a-chip provided by the present invention can be used for high-throughput vascularized tissue construction and has broad application prospects in in vitro model construction, drug screening, regenerative medicine and other fields. Attached Figure Description

[0025] Figure 1 This is a schematic diagram of the layered structure of the organ-on-a-chip construction device in an embodiment of the present invention, and a partially enlarged view thereof;

[0026] Figure 2 This is a schematic diagram of the layered structure and preparation process of the channel layer in an embodiment of the present invention;

[0027] Figure 3 This is a flowchart illustrating the construction process of an open vascularized organ-on-a-chip in an embodiment of the present invention;

[0028] Figure 4 This is a fluorescence micrograph of vascularized tissue constructed using an organ-on-a-chip construction device in an embodiment of the present invention. Detailed Implementation

[0029] The technical solution of the present invention will now be clearly and completely described with reference to specific embodiments and accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0030] In the description of this invention, the use of terms such as "upper," "lower," "inner," "outer," "left," "right," "top," and "bottom," indicating orientation or positional relationships, is based on the orientation or positional relationships shown in the accompanying drawings. These terms are used solely for the convenience of describing the invention and for 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 limitations on the invention. The terms "first," "second," etc., in the specification, claims, and accompanying drawings of this invention are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence. It should be understood that such terms can be used interchangeably where appropriate; this is merely a way of distinguishing objects with the same attributes in the description of embodiments of the invention. Furthermore, the terms "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusion, but rather to include other units not explicitly listed or inherent to these products or devices.

[0031] like Figure 1 As shown, this embodiment of the invention provides a device for constructing open vascularized organ-on-a-chip (hereinafter referred to as "organ-on-a-chip construction device" or "device"), which mainly consists of three functional layers with separate designs: a culture medium chamber layer 1, a channel layer 2, and a base layer 3. The three-layer structure can be bonded together by a pressure-sensitive double-sided adhesive with good biocompatibility.

[0032] In this embodiment, the device has eight repeating units, also known as culture units, arranged in an array. Each culture unit has one channel unit and four culture medium chambers. However, in other embodiments, the number and arrangement of culture units on the organ-on-a-chip fabrication device can be set as needed; there can be one or more culture units. The cross-sectional shape and arrangement of the culture medium chambers within the culture unit can also be designed according to requirements, typically with a circular cross-section. Furthermore, the shape, structure, and arrangement of the culture medium chambers in multiple culture units within the same device are not required to be completely identical. Each culture unit within the same device can serve the same or different functions; for example, different length and width dimensions of vascularized tissue can be constructed in different culture units.

[0033] like Figure 1As shown, in this embodiment, channel layer 2 includes eight channel units. Taking one channel unit as an example, it mainly has two channel structures: a matrix gel channel 21 and a culture medium channel 22. The matrix gel channel 21 adopts an open design, unlike the closed channel design. The open channel has no sidewalls and its sides are connected to the outside. Utilizing the capillary flow of matrix gel in the open channel, the matrix gel can be confined within the open channel, i.e., the matrix gel channel 21. There are two culture medium channels 22, respectively located on both sides of the matrix gel channel 21 and connected to it. Furthermore, at the two ends of the culture medium channel 22 away from the matrix gel channel 21, there are circular holes with a diameter larger than the width of the culture medium channel, i.e., culture medium inlet / outlet ports 23 (referred to as "culture medium inlet / outlet"). After the channel layer 2 is assembled with the culture medium chamber layer 1, the culture medium inlet / outlet 23 is arranged correspondingly to the culture medium chamber 11 in the culture medium chamber layer 1 and the two are connected. The shape and size of the culture medium inlet / outlet 23 are basically the same as the cross-sectional shape of the culture medium chamber 11, usually a circular hole.

[0034] In channel layer 2, the matrix gel channel 21 and the culture medium channel 22 have a certain height difference, typically 100-500 μm. This height difference, along with the wetting properties of the liquid, confines the matrix gel within the channels. For example... Figure 3As shown, the channel layer is formed by hot-pressing two laser-cut acrylic sheets. The thickness of each acrylic sheet is 100-500 μm. The thickness of the two acrylic sheets can be the same or different, depending on the height difference requirements of the two channels. The two acrylic sheets have certain structural differences. The lower acrylic sheet can be called "substrate 2a", and its thickness is usually 100-500 μm; the upper acrylic sheet can be called "cover sheet 2b", and its thickness is usually 100-500 μm. In this embodiment, the hollowed-out area on the substrate 2a has an axisymmetric structure. The central part, i.e., the axial region, is used to form the matrix adhesive channel 21. The regions on both sides of the axial region are connected to form the culture medium channel 22 and the culture medium inlet / outlet 23. The cover sheet 2b also has an axisymmetric structure, but the difference is that the hollowed-out area is only used to form the culture medium channel 22 and the culture medium inlet / outlet 23. These two parts, together with the hollowed-out area at the corresponding position of the substrate 2a, constitute the culture medium channel 22 and the culture medium inlet / outlet 23. After the substrate 21 and cover plate 22 are heat-sealed together, a channel layer 2 is formed, creating an open matrix adhesive channel 21 and two culture medium channels 22 located on either side of the matrix adhesive channel 21. After the channel layer 2 is fixedly connected to the base layer 3, the matrix adhesive channel 21 is open on its sides and closed at the top and bottom; the culture medium channels 22 are only open on their upper surface and the side adjacent to the matrix adhesive channel 21. The height of the matrix adhesive channel 21 is equal to the thickness of the substrate 2a, and the height of the culture medium channel 2 is the sum of the thicknesses of the substrate 2a and the cover plate 2b, thus creating a height difference between the matrix adhesive channel 21 and the culture medium channel 22. Besides acrylic, in other embodiments, the channel layer can also be made of materials with good biocompatibility and easy processing properties, such as polycarbonate.

[0035] The culture medium chamber layer 1 is mainly used to contain the culture medium. In this embodiment, the culture medium chamber layer 1 adopts a through-hole array structure, that is, it has 32 circular through holes arranged in an array. The circular through holes of the culture medium chamber layer 1 correspond one-to-one with the circular culture medium inlet / outlet 23 of the channel layer 2, so that one culture medium channel 22 of the channel layer 2 is connected to two culture medium chambers 11 in the culture medium chamber layer 1. The culture medium chamber layer 1 can be made of materials with good biocompatibility, such as polycarbonate or acrylic, and is prepared by CNC machining or laser cutting, with a thickness of 1-5 cm.

[0036] Understandably, after the culture medium chamber layer 1 and the channel layer 2 are bonded together with pressure-sensitive double-sided adhesive, the culture medium channel 22 changes from an open structure when the matrix adhesive is added to a closed structure after sealing, thereby enabling the flow of culture medium inside the channel 22 along the side wall of the matrix adhesive.

[0037] The base layer 3 can be made of biocompatible materials such as glass and quartz with a visible light transmittance of more than 90%. For example, commonly used microscope slides or microscope coverslips can be used. It is mainly used to seal the bottom of the chip and facilitate microscopic imaging of the sample.

[0038] It is understood that the above-described structural design of the culture unit is only one option, and various designs can be made in other embodiments. For example, the culture unit is not required to be an axisymmetric structure and can be any shape; the number of culture medium channels 22 is also not limited, and can be several surrounding and connected to the matrix gel channel 21, as long as they can be connected to the matrix gel channel 21 and form a height difference; the number of culture medium inlets / outlets 23 connected to the culture medium channel 22 is also not limited, and can be designed in two or more according to the shape of the channel. Generally, they should be the same as the number of culture medium chambers in the culture medium chamber layer 1, with basically the same shape, and arranged corresponding to the culture medium chambers. In other words, the structure of the culture unit generally only needs to meet the internal channel design requirements, especially the open design of the matrix gel channel 21, the height difference between the matrix gel channel 21 and the culture medium channel 22, and the connection of two or more culture medium inlets / outlets 23 to the same culture medium channel 22.

[0039] It is worth noting that in this embodiment, channel layer 2 is obtained by heat-sealing a laser-cut substrate and a cover plate. This method is simple to operate and has a low cost. In other embodiments, channel layer 2 may not be layered and may be integrally formed by 3D printing, injection molding, or other methods.

[0040] In this embodiment, the biocompatible pressure-sensitive double-sided adhesive is mainly used to achieve bonding after chip separation. As a preferred solution, the pressure-sensitive double-sided adhesive has the advantages of biocompatibility, no leakage risk, convenient operation, and reliable bonding.

[0041] Another embodiment of the present invention discloses the application of the organ-on-a-chip construction apparatus described in the above embodiments in the construction of vascularized tissue. For example... Figure 3 As shown, this method for constructing a split, open-system vascularized organ-on-a-chip mainly includes the following steps:

[0042] S1: Sterilize the organ-on-a-chip device with ultraviolet irradiation or ethylene oxide, and assemble the relevant parts.

[0043] Understandably, before practical application, the device has a split structure, consisting of three parts: culture medium chamber layer 1, base layer 3, and channel layer 2. In practical application, channel layer 2 is usually formed by first sealing the cover plate 2a and the substrate 2b together by heat pressing, and then the channel layer 2 and the base layer 3 are bonded together with pressure-sensitive double-sided adhesive to form a whole.

[0044] S2: Endothelial cells, stromal cells and other parenchymal cells (optional) are resuspended in a matrix gel solution such as fibrin or collagen, and then added into the matrix gel channel 21 of channel layer 2. The matrix gel is confined within the open matrix gel channel 21 by the capillary flow of the matrix gel in the open channel.

[0045] S3: After the matrix adhesive has cured, the culture medium chamber layer 1 is bonded to the channel layer 2 using a biocompatible pressure-sensitive double-sided adhesive, ensuring that the culture medium chamber 11 of the culture medium chamber layer 1 corresponds one-to-one with the culture medium inlet / outlet 23 at both ends of the culture medium channel 21 of the channel layer 2, and that the two are connected.

[0046] S4: Different volumes of culture medium are added to the culture medium chambers 11 on both sides of the matrix gel channel 21, generating a hydrostatic pressure difference on both sides of the matrix gel, thereby creating interstitial flow in the matrix gel and promoting angiogenesis.

[0047] Optionally, in some applications, such as constructing a complete endothelial cell barrier on the sidewall of the matrix gel, it is also necessary to generate channel flow along the sidewall of the matrix gel. Therefore, a rocker-type shaker can be used to apply a tilting oscillation parallel to the channel direction, so that there is a liquid level difference between the two culture medium chambers at both ends of the culture medium channel 22, thereby realizing the formation of channel flow in the culture medium channel (i.e., along the sidewall of the matrix gel).

[0048] Furthermore, the present invention also provides a specific example of applying the organ-on-a-chip construction device described in the above embodiments to the evaluation of anti-angiogenic drugs.

[0049] Two acrylic sheets with eight channel units were cut from a 500μm thick acrylic sheet using a laser cutting instrument. The two acrylic sheets were then heat-pressed together to form a channel layer. Pressure-sensitive double-sided adhesive cut by a die-cutting machine was used to bond the sealed acrylic sheets to a rectangular coverslip to form a whole. The whole structure formed by the base layer and the channel layer, as well as the 5cm high acrylic culture medium chamber layer machined by CNC, were then sterilized with ethylene oxide for later use.

[0050] Subsequently adopted as follows Figure 4 The experimental procedure shown involves resuspending endothelial cells and stromal cells in a matrix gel solution with fibrinogen as the main component, mixing it with thrombin, and then rapidly adding it to fill the matrix gel channels in the channel layer. Thrombin is used to promote the solidification of the matrix gel solution. Then, pressure-sensitive double-sided tape cut by a die-cutting machine is used to bond the culture medium chamber layer and the channel layer together. Different volumes of culture medium are then added to the culture medium chambers on the left and right sides of the matrix gel that are connected to the culture medium channels to achieve a hydrostatic pressure difference across the matrix gel, thereby driving the interstitial flow within the matrix gel and promoting the assembly of endothelial cells to form a vascular network.

[0051] Meanwhile, bevacizumab at concentrations of 0, 100, 200, 400, 600, 800, 1000, and 2000 μg / ml were added to eight culture units, respectively. After 7 days of culture, microscopic images of the vascular networks formed under different conditions were acquired using fluorescence confocal microscopy. Structural information such as the length, area, and number of nodes of the vascular networks were statistically analyzed to evaluate the anti-angiogenic efficacy of bevacizumab.

[0052] Furthermore, the present invention also provides a specific example of applying the open vascularization device described in the above embodiments to the construction of vascularized liver tissue.

[0053] An acrylic sheet capable of forming a repeating channel unit structure was cut from a 100μm thick acrylic sheet using a laser cutting instrument. Then, two cut acrylic sheets were heat-pressed together to form a channel layer. The acrylic sheet cut with double-sided adhesive was then glued onto a rectangular glass slide to form a whole. The whole structure formed by the base layer and the channel layer, as well as the CNC-machined 1cm high polycarbonate culture medium chamber layer, were then sterilized under ultraviolet light for 12 hours for later use.

[0054] Then proceed as follows Figure 4 The experimental procedure shown involves resuspending endothelial cells, stromal cells, and parenchymal cells (primarily composed of primary hepatocytes) in fibrinogen solution, mixing them with thrombin, and then rapidly adding the mixture to fill the matrix gel channels in the channel layer. Pressure-sensitive double-sided tape, cut by a die-cutting machine, is then used to bond the culture medium chamber layer and the channel layer together. Different volumes of culture medium are then added to the culture medium chambers on both sides of the matrix gel, which connect to the culture medium channels, creating a hydrostatic pressure difference across the matrix gel. This drives interstitial flow within the matrix gel, promoting angiogenesis.

[0055] After approximately 7 days of culture, vascularized tissue containing primary hepatocytes was obtained. This vascularized tissue can be further used for research such as drug hepatotoxicity evaluation and drug metabolism testing.

[0056] Finally, it should be noted that although the embodiments of the present invention have been described above in conjunction with the accompanying drawings, the present invention is not limited to the specific embodiments and application fields described above. The specific embodiments described above are merely illustrative and instructive, and not restrictive. Those skilled in the art, guided by this specification, can make many other forms without departing from the scope of protection of the claims of the present invention, and all of these are within the scope of protection of the present invention.

Claims

1. A device for constructing open-system vascularized organ-on-a-chip, characterized in that, include: basal layer; The channel layer has at least one channel unit; the channel unit has a first channel with one side open and two or more second channels connected to the side of the first channel, the second channel has two or more culture medium inlets and outlets, and there is a preset height difference between the second channel and the first channel; The culture medium chamber layer has multiple culture medium chambers; The base layer, channel layer and culture medium chamber layer are designed separately. When stacked from bottom to top, the culture medium chambers of the culture medium chamber layer correspond one-to-one with the culture medium inlet and outlet of the second channel of the channel layer and are connected.

2. The apparatus as claimed in claim 1, characterized in that, The channel layer is formed by hot-pressing a substrate and a cover plate together; the substrate and the cover plate are respectively provided with hollow structures, which are used to form the first channel and the second channel; The height of the first channel is determined by the thickness of the substrate, and the height of the second channel is determined by the sum of the thicknesses of the substrate and the cover plate. The thickness of the substrate and cover plate does not need to be the same.

3. The apparatus as described in claim 2, characterized in that, The thickness of the substrate and cover plate is 100-500μm, including the endpoint values; the height of the first channel is less than the height of the second channel, and the preset height difference is 100-500μm.

4. The apparatus according to any one of claims 1 to 3, characterized in that, The channel unit has two second channels, which are respectively arranged on the side of the first channel; the second channel is provided with two culture medium inlets and outlets, which are respectively arranged at the ends of the second channel away from the first channel.

5. The apparatus according to any one of claims 1 to 3, characterized in that, The number of culture medium chambers in the culture medium chamber layer is consistent with the number of culture medium inlets and outlets in the channel layer; the height of the culture medium chamber is 1-5cm; the cross-section of the culture medium chamber is circular; the culture medium inlet and outlet are circular holes, and the diameter of the circular holes is the same as the diameter of the circular cross-section of the culture medium chamber.

6. The apparatus according to any one of claims 1 to 3, characterized in that, The channel layer is made of acrylic or polycarbonate; the culture medium chamber layer is made of acrylic / polycarbonate; the base layer is made of a biocompatible material with a visible light transmittance greater than 90%; preferably glass or quartz.

7. The apparatus according to any one of claims 1 to 3, characterized in that, When the device is in use, the base layer, channel layer, and culture medium chamber layer are bonded and fixed together by pressure-sensitive double-sided adhesive.

8. A method for constructing an open-system vascularized organ-on-a-chip, characterized in that, The apparatus for constructing open-system vascularized organ-on-a-chip according to claims 1 to 7 comprises: The base layer and the channel layer are fixedly connected, and the first channel is closed from top to bottom after the connection. A matrix adhesive solution is injected into the first channel, and the matrix adhesive is confined within the first channel; After the matrix adhesive has cured, the culture medium chamber layer and the channel layer are fixedly connected, and the culture medium chamber and the culture medium inlet and outlet of the second channel are connected one by one. Culture medium is added to the culture medium chamber, and a hydrostatic pressure difference is generated between the sides of the matrix gel, which promotes interstitial flow within the matrix gel.

9. The method for constructing an open-system vascularized organ-on-a-chip as described in claim 8, characterized in that, Also includes: By oscillating, a liquid level difference is created between two or more culture medium chambers connected by each second channel, so as to form a channel flow along the sidewall of the matrix gel within the second channel.

10. An open-system vascularized organ-on-a-chip, characterized in that, Constructed by the method for constructing open vascularized organ-on-a-chip as described in claim 8 or 9.

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