Organ-like culture chip
By setting up injection channels and semi-permeable membranes in microfluidic chips, the blood-brain barrier function of brain-like organ models can be simulated, which solves the problem of insufficient functional simulation of brain-like organ models in the prior art and improves the accuracy of drug screening and evaluation.
Patent Information
- Application Number
- CN202422872639.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-25
- Publication Date
- 2026-01-02
- Estimated Expiration
- 2034-11-25
AI Technical Summary
Existing technologies are unable to effectively simulate the blood-brain barrier function of brain-like organs, resulting in insufficient accuracy in drug screening and evaluation.
Design an organoid culture chip that simulates the blood-brain barrier function of brain-like organoid models by setting up injection channels and semi-permeable membranes in a microfluidic chip, and uses blood simulants to exchange substances with organoid cells.
It improves the accuracy of drug screening and evaluation, simulates the blood-brain barrier function of brain-like organs, and enhances the authenticity and effectiveness of drug detection.
Smart Images
Figure CN223752809U_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The utility model relates to micro -fluidic chip technical field especially, it relates to a kind of organoid culture chip. BACKGROUND
[0002] Organoid is the three-dimensional cell complex that is induced differentiation to stem cell or organ cell using 3D culture technique in vitro and is similar to target organ or tissue in structure and function, it has stable phenotype and genetic characteristics, can be long-term cultured in vitro.Organoid can simulate the genetic characteristics and epigenetic characteristics of target tissue or organ to a great extent, has wide application prospect in organ development, precision medicine, regenerative medicine, drug screening, gene editing, disease modeling and other fields.
[0003] Before drug enters clinical test, drug screening and evaluation are the core link of drug development.Currently commonly used two-dimensional in vitro cell culture model and animal model, in predicting the sensitivity of human pathophysiology or specific patient to drug and drug toxicity, there are great defects.In order to reduce the cost of drug development, the effectiveness of preclinical drug evaluation must be improved, and invalid candidate drugs are eliminated as soon as possible.The emergence of microfluidic organ chip technology is expected to solve many problems encountered in conventional drug screening and evaluation.Microfluidic technology can reconstruct the complex physiological environment in vivo under micron scale, and the constructed bionic organ chip model can more truly reflect the possible action of drug on tissue / organ, thereby improving the accuracy of drug screening and evaluation.
[0004] From the early organ chip prototype, lung, intestine, liver and other single organ chip to the multi-organ chip in series, organoid and organ chip have made great progress. Brain-like organ model is a brain-like tissue induced by pluripotent stem cells, which has various cell types and structural functional domains of brain, can partially reproduce the structure and function of brain tissue, and even the disease occurrence process. Brain-like organ technology is a frontier technology in the field of brain science and stem cells and regenerative medicine, how to realize the blood-brain barrier function simulation of brain-like organ model by using microfluidic chip has become a problem to be solved in the industry. UTILITARY MODEL CONTENT
[0005] The utility model provides a kind of organoid culture chip, to realize the blood vessel barrier function simulation of brain-like organ model by using microfluidic chip.
[0006] The utility model provides a kind of organoid culture chip, including first substrate and second substrate, the first substrate is stacked with the second substrate;The first substrate is equipped with first cavity, and the first cavity is used to accommodate organoid cell, and the second substrate is equipped with second cavity, and the second cavity is communicated with the first cavity, and the second cavity is used to accommodate cell culture medium;The periphery of the second cavity is equipped with liquid injection flow channel, and the liquid injection flow channel is used to flow detection drug and blood analog, and semi-permeable membrane is equipped between the liquid injection flow channel and the second cavity to carry out material exchange.
[0007] According to the organoid culture chip provided by the utility model, the liquid injection flow channel includes a pair of oppositely arranged flow channels, each of the flow channels includes: a first flow channel, a second flow channel and a third flow channel connected in sequence, the second flow channel is a circular arc-shaped flow channel, a pair of the second flow channels are arranged around the periphery of the second cavity, and the semi-permeable membrane is arranged between the second flow channel and the second cavity.
[0008] According to the organoid culture chip provided by the utility model, each of the second flow channels includes a plurality of branch flow channels, each of the branch flow channels is circular arc-shaped, the first ends of the plurality of branch flow channels are communicated with the first flow channel, and the second ends of the plurality of branch flow channels are communicated with the third flow channel.
[0009] According to the organoid culture chip provided by the utility model, the second substrate is also provided with a cell culture medium flow channel, and the cell culture medium flow channel is communicated with the second cavity.
[0010] According to the organoid culture chip provided by the utility model, the liquid injection flow channel and the second cavity are configured as a blood-brain barrier unit, and a plurality of the blood-brain barrier units are arranged on the second substrate.
[0011] According to the organoid culture chip provided by the utility model, the third substrate is stacked with the second substrate, a plurality of first inlets and a plurality of first outlets are arranged on the third substrate, a plurality of the liquid injection flow channels are divided into a plurality of groups, the plurality of liquid injection flow channels in each group are sequentially communicated, and are communicated with one first inlet and one first outlet.
[0012] According to the organoid culture chip provided by the utility model, a plurality of second inlets and a second outlet are arranged on the second substrate, a plurality of the second cavities are divided into a plurality of groups, the plurality of second cavities in each group are sequentially communicated, and are communicated with one second inlet and the second outlet.
[0013] According to the organ-like culture chip, multiple blood-brain barrier units form a rhombic structure, multiple first inlets are located at one side of one corner of the rhombic structure, and multiple first outlets are oppositely arranged with the remaining three corners of the rhombic structure one by one.
[0014] According to the organ-like culture chip, multiple blood-brain barrier units form a rhombic structure, multiple second inlets are oppositely arranged with one corner of the rhombic structure, and the second outlet is located at the center of the rhombic structure.
[0015] According to the organ-like culture chip, multiple second inlets are used for injecting substances for regulating cell growth into the second cavity.
[0016] The organ-like culture chip provided by the utility model realizes the application of the micro-fluidic chip in the brain-like organ by arranging the liquid injection flow channel around the second cavity and arranging the semi-permeable membrane between the second cavity and the liquid injection flow channel, so that the brain-like organ cells and the blood analogues exchange substances through the semi-permeable membrane to simulate the blood-brain barrier. BRIEF DESCRIPTION OF DRAWINGS
[0017] In order to more clearly illustrate the technical solutions of the utility model or the prior art, the following will briefly introduce the drawings needed to be used in the embodiments or the prior art description. Obviously, the drawings in the following description are some embodiments of the utility model, and for those skilled in the art, other drawings can also be obtained without creative labor on the basis of these drawings.
[0018] Figure 1 It is the structure schematic view of the organ-like culture chip provided by the utility model.
[0019] Figure 2 It is Figure 1 The longitudinal section view of the blood-brain barrier unit shown in the figure.
[0020] Figure 3 It is Figure 1 The transverse section view of the blood-brain barrier unit shown in the figure.
[0021] Reference signs:
[0022] 1, first substrate; 2, second substrate; 11, first cavity; 21, second cavity; 22, liquid injection flow channel; 23, semi-permeable membrane; 24, cell culture medium flow channel; 25, first inlet; 26, first outlet; 27, second inlet; 28, second outlet; 200, blood-brain barrier unit; 221, first flow channel; 222, second flow channel; 223, third flow channel; 2221, branch flow channel. DETAILED DESCRIPTION
[0023] In order to make the purpose, technical scheme and advantages of the utility model more clear, the technical scheme in the utility model will be described clearly and completely below in combination with the drawings in the utility model. Obviously, the described embodiments are part of the embodiments of the utility model, rather than all the embodiments. Based on the embodiments in the utility model, all other embodiments obtained by the person skilled in the art without creative labor belong to the protection scope of the utility model.
[0024] The utility model discloses an organoid culture chip. Figures 1-3 The utility model discloses an organoid culture chip.
[0025] As Figure 2 shown, in the embodiment of the utility model, organoid culture chip includes: first substrate 1 and second substrate 2, first substrate 1 is overlaid with second substrate 2, and first substrate 1 is equipped with first cavity 11, and first cavity 11 is used to hold organoid cell, and second substrate 2 is equipped with second cavity 21, and second cavity 21 communicates with first cavity 11, and second cavity 21 is used to hold cell culture medium. The periphery of second cavity 21 is equipped with liquid injection flow channel 22, and liquid injection flow channel 22 is used to flow detection drug and blood analog, and semi-permeable membrane 23 is equipped between liquid injection flow channel 22 and second cavity 21 to exchange material.
[0026] Specifically, organoid cell is placed in first cavity 11, and cell culture medium is injected in second cavity 21 to culture organoid cell. Blood analog flows along liquid injection flow channel 22, and blood analog exchanges material with organoid cell in second cavity 21 through semi-permeable membrane 23 to control drug or nutrient supply of organoid culture medium and blood vessel model by organoid culture chip, simulate blood-brain barrier. Detection drug flows to detection equipment by liquid injection flow channel 22 to detect.
[0027] In the embodiment, the bottom of first cavity 11 is nanofiber mesh scaffold bottom, and first cavity 11 is cylindrical, which is conducive to organoid cell culture, aims at increasing the attachment point of organoid cell and reusability.
[0028] Further, the injection channel 22 includes an inlet channel, an intermediate channel, and an outlet channel connected in sequence. Optionally, in one embodiment of the present invention, the inlet channel and the outlet channel can be arranged parallel to each other, and the intermediate channel is a similar annular channel. The intermediate channel surrounds the second cavity 21 so that the blood simulant in the intermediate channel can exchange substances with the organoid cells in the second cavity 21. Optionally, in another embodiment of the present invention, the inlet channel, the intermediate channel, and the outlet channel constitute a channel. The injection channel 22 includes a pair of channels arranged opposite to each other. The intermediate channel in each channel is an arc-shaped channel. The pair of arc-shaped channels surround the two sides of the second cavity 21 so that the blood simulant in the intermediate channel can exchange substances with the organoid cells in the second cavity 21.
[0029] Optionally, in embodiments of this invention, the organoid cells are brain microvascular endothelial cells, embryonic stem cells (ESCs), and induced pluripotent stem cells (iPSCs), which are used as cultured cells for research. These cells constitute the main barrier of the blood-brain barrier, are highly selective, and can restrict certain substances from entering the brain from the blood.
[0030] The organoid culture chip provided in this embodiment of the invention allows organoid cells and blood simulants to exchange substances through a semipermeable membrane by setting a liquid injection channel around the second cavity and setting a semipermeable membrane between the second cavity and the liquid injection channel, thereby simulating the blood-brain barrier and realizing the application of microfluidic chips in brain-like organs.
[0031] like Figure 3 As shown, in an embodiment of this utility model, the injection channel 22 includes a pair of oppositely arranged channels. Each channel includes a first channel 221, a second channel 222, and a third channel 223 connected in sequence. The second channel 222 is an arc-shaped channel. The pair of second channels 222 surround the second cavity 21. The semi-permeable membrane 23 is disposed between the second channel 222 and the second cavity 21.
[0032] Specifically, the blood simulant enters the second channel 222 from the first channel 221. In the second channel 222, the blood simulant exchanges substances with the organoid cells in the second cavity 21 through the semipermeable membrane 23 to simulate the blood-brain barrier.
[0033] like Figure 2 As shown, each second flow channel 222 includes multiple branch flow channels 2221, each branch flow channel 2221 is arc-shaped, the first end of the multiple branch flow channels 2221 is connected to the first flow channel 221, and the second end of the multiple branch flow channels 2221 is connected to the third flow channel 223.
[0034] Specifically, the second flow channel 222 is designed as a plurality of branch flow channels 2221, aiming to simulate the real blood vessel distribution state, the blood vessel thickness, and increase the contact area between the blood simulation and the organoid cells.
[0035] As shown in the embodiment of the utility model, the second substrate 2 is further provided with a cell culture medium flow channel 24, and the cell culture medium flow channel 24 is in communication with the second cavity 21. Figure 2 The cell culture medium flow channel 24 also includes an inlet flow channel and an outlet flow channel, both of which are in communication with the second cavity 21, and the cell culture medium enters the second cavity 21 from the inlet flow channel and flows out from the outlet flow channel.
[0036] As shown in the embodiment of the utility model, the second substrate 2 is further provided with a cell culture medium flow channel 24, and the cell culture medium flow channel 24 is in communication with the second cavity 21. Figure 1
[0037] Further, the organoid culture chip further includes a third substrate, the third substrate is stacked with the second substrate 2, and the third substrate is provided with a first inlet 25, a second inlet 27, a first outlet 26 and a second outlet 28.
[0038] Specifically, in the embodiment, the organoid culture chip is a three-layer structure, and the first substrate 1, the second substrate 2 and the third substrate are stacked from bottom to top. The first substrate 1 is provided with a first cavity 11, the second substrate 2 is provided with a plurality of blood-brain barrier units 200, and the third substrate encapsulates the plurality of blood-brain barrier units 200. The third substrate is provided with a first inlet 25, a second inlet 27, a first outlet 26 and a second outlet 28 to communicate with the plurality of blood-brain barrier units 200, and inject cell culture medium and blood simulation into the blood-brain barrier units 200.
[0039] Specifically, the third substrate is provided with a plurality of first inlets 25 and first outlets 26, and the plurality of injection flow channels 22 are divided into a plurality of groups. The plurality of injection flow channels 22 in each group are sequentially connected and communicated with one first inlet 25 and one first outlet 26.
[0040] As described above, the injection flow channel 22 includes a pair of flow channels, for the convenience of description, the pair of flow channels are respectively named as flow channel A and flow channel B. The plurality of flow channels A in each group are sequentially connected, and the plurality of flow channels B in each group are sequentially connected. After the blood simulation enters from the first inlet 25, it flows to the flow channel A and the flow channel B, and then flows out from the first outlet 26. During the flow of the blood simulation, the blood simulation exchanges substances with the organoid cells in the second flow channel 222 of each injection flow channel 22 and the second cavity 21.
[0041] As shown in the embodiment of the utility model, the second substrate 2 is further provided with a cell culture medium flow channel 24, and the cell culture medium flow channel 24 is in communication with the second cavity 21. Figure 1 In the shown embodiment, the plurality of blood brain barrier units 200 form a rhombus structure, the plurality of first inlets 25 are located on one side of one corner of the rhombus structure, and the plurality of first outlets 26 are respectively arranged opposite to the remaining three corners of the rhombus structure.
[0042] Specifically, in the embodiment, the three liquid injection channels 22 on the upper left side are sequentially communicated with the first outlet 26 located in the uppermost position in the middle; the three liquid injection channels 22 on the lower left side are sequentially communicated with the first outlet 26 located in the lowermost position in the middle; and the three liquid injection channels 22 on the right side are sequentially communicated with the first outlet 26 located on the right side in the middle. Figure 1 Figure 1 Figure 1
[0043] In the embodiment shown in FIG. 1, the number of first inlets 25 is four, one of which is a standby inlet, and the remaining three first inlets 25 are used in cooperation with one first outlet 26.
[0044] It can be understood that in the embodiment of the utility model, by grouping the plurality of liquid injection channels 22, the plurality of liquid injection channels 22 in each group are sequentially communicated, which can reduce the number of microchannels on the second substrate 2, and in addition, multiple blood brain barrier simulations can be realized at the same time. Further, the structure formed by the plurality of blood brain barrier units 200 can also be other, such as a regular hexagonal structure, and is not limited to the rhombus structure described in the embodiment of the utility model.
[0045] The third substrate is further provided with a plurality of second inlets 27 and a second outlet 28, and the plurality of second cavities 21 are divided into a plurality of groups, the plurality of second cavities 21 in each group are sequentially communicated, and are communicated with one second inlet 27 and the second outlet 28.
[0046] Specifically, the plurality of second cavities 21 in each group are sequentially communicated through the cell culture medium channel 24 and connected with one second inlet 27. The cell culture medium is injected from the respective second inlet 27, then enters each second cavity 21 along the cell culture medium channel 24 of each group, and flows out from the second outlet 28.
[0047] When the plurality of blood brain barrier units 200 form a rhombus structure, the plurality of second inlets 27 are respectively arranged opposite to one corner of the rhombus structure, and the second outlet 28 is located at the center of the rhombus structure. Specifically, three second cavities 21 above the second outlet 28 are communicated and then communicated with the second inlet 27 above the second outlet 28; three second cavities 21 below the second outlet 28 are communicated and then communicated with the second inlet 27 below the second outlet 28; the second cavity 21 on the left side of the second outlet 28 is communicated with the second inlet 27 on the left side of the second outlet 28; and the second cavity 21 on the right side of the second outlet 28 is communicated with the second inlet 27 on the right side of the second outlet 28.
[0048] Alternatively, the grouping of the plurality of second cavities 21 can also be other, such as three second cavities 21 on the left side of the second outlet 28 as a group, communicated with the second inlet 27 on the left side; three second cavities 21 on the right side of the second outlet 28 as a group, communicated with the second inlet 27 on the right side; and the second cavities 21 above and below the second outlet 28 are respectively a group, communicated with the respective adjacent second inlet 27.
[0049] Further, as shown in Figure 1 The number of second inlets 27 above the second outlet 28 is multiple, and the remaining second inlets 27 are used to inject growth factors such as CK-MB, GST-α, TGF-β1 into the second cavity 21.
[0050] Finally, it should be noted that: the above examples are only used to illustrate the technical solutions of the present application, and not to limit it; although the present application has been described in detail with reference to the foregoing examples, those skilled in the art should understand that: it can still modify the technical solutions recorded in the foregoing examples, or make equivalent replacement for part of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present application.
Claims
1. An organoid culture chip, characterized by, The device comprises a first substrate and a second substrate, and the first substrate is stacked with the second substrate; The first substrate is provided with a first cavity for accommodating organoid cells, and the second substrate is provided with a second cavity in communication with the first cavity for accommodating cell culture medium; The second cavity is provided with a liquid injection channel for flowing detection drugs and blood analogues, and a semi-permeable membrane is arranged between the liquid injection channel and the second cavity for material exchange.
2. The organoid culture chip of claim 1, wherein, The liquid injection channel comprises a pair of oppositely arranged channels, each of which comprises a first channel, a second channel and a third channel in sequence, the second channel is a circular arc channel, and a pair of second channels are arranged around the second cavity, and the semi-permeable membrane is arranged between the second channel and the second cavity.
3. The organoid culture chip of claim 2, wherein, Each of the second channels comprises a plurality of branch channels, each of which is a circular arc type, the first ends of the plurality of branch channels are in communication with the first channel, and the second ends of the plurality of branch channels are in communication with the third channel.
4. The organoid culture chip of claim 1, wherein, The second substrate is also provided with a cell culture medium channel in communication with the second cavity.
5. The organoid culture chip of claim 4, wherein, The liquid injection channel and the second cavity are configured as a blood-brain barrier unit, and the second substrate is provided with a plurality of blood-brain barrier units.
6. The organoid culture chip of claim 5, wherein, The device further comprises a third substrate stacked with the second substrate; The third substrate is provided with a plurality of first inlets and a plurality of first outlets, and a plurality of liquid injection channels are divided into a plurality of groups, and the liquid injection channels in each group are in communication with each other and with one first inlet and one first outlet.
7. The organoid culture chip of claim 6, wherein, The second substrate is also provided with a plurality of second inlets and a second outlet, and a plurality of second cavities are divided into a plurality of groups, and the second cavities in each group are in communication with each other and with one second inlet and the second outlet.
8. The organoid culture chip of claim 6, wherein, The plurality of blood-brain barrier units form a rhombus structure, the plurality of first inlets are located on one side of one corner of the rhombus structure, and the plurality of first outlets are respectively arranged opposite to the remaining three corners of the rhombus structure.
9. The organoid culture chip of claim 7, wherein, The plurality of blood-brain barrier units form a rhombus structure, the plurality of second inlets are respectively arranged opposite to one corner of the rhombus structure, and the second outlet is located at the center of the rhombus structure.
10. The organoid culture chip of claim 7, wherein, The plurality of second inlets are used to inject substances for regulating cell growth into the second cavities.