CN113621515B

CN113621515B - Glomerular chip

Info

Publication number: CN113621515B
Application number: CN202110919100.4A
Authority: CN
Inventors: 丁卫平; 戴志琳; 李成盼
Original assignee: University of Science and Technology of China USTC
Current assignee: University of Science and Technology of China USTC
Priority date: 2021-08-11
Filing date: 2021-08-11
Publication date: 2023-06-02
Anticipated expiration: 2041-08-11
Also published as: CN113621515A

Abstract

The invention discloses a glomerular chip, which comprises a first chip, a second chip, a third chip and a fourth chip which are stacked from top to bottom, wherein the glomerular chip can simulate an external glomerular capillary vessel structure, can realize separated capillary vessel clusters and abalone cavities, provides a solution idea for the problem that the current glomerular model has low bionic degree, and is based on the glomerular chip, a hollow fiber bundle is packaged in the abalone cavities of the chip in a 'C' shape mode, and foot cells and endothelial cells are respectively inoculated at two sides of a fiber tube, so that the glomerular capillary vessel structure is simulated in vitro, the separated glomerular capillary vessel clusters and the abalone cavities are realized, a filtrate outlet on the chip simulates blood filtering behavior in a human body, and the physiological similarity of the glomerular model is improved.

Description

Glomerular chip

Technical Field

The invention relates to the technical field of organ chips, in particular to a glomerular chip.

Background

Kidneys are vital organs of the human body, and have extremely complex structures and microenvironments. Kidneys consist of millions of nephrons, with glomeruli being an important component of the nephron. The glomerulus is responsible for performing a hemofiltration function, whose filtration membrane is of a three-layer structure with a basement membrane in the middle. Glomerular filtration is the initial process of urine formation, as blood flows through the glomeruli, most of the material is filtered by glomerular capillaries and enters the chamber of the baud, forming raw urine. Studies have shown that most kidney diseases (e.g., chronic kidney disease, diabetic nephropathy, and nephrotic syndrome) are caused by glomerular dysfunction. Thus, researchers have focused on building in vitro glomerular models to reveal the mechanisms of glomerular function.

Currently, there are two main types of in vitro glomerular models: kidney organoids and glomerular chips. Glomeruli constructed by kidney organoids can express podocyte-related proteins and have glomerular filtration function. However, the existing kidney organoids have not achieved a spherical glomerular structure with a bowing cavity, and lack a perfusable vascular network, so their application is limited. Glomerular chips are in vitro glomerular models built based on emerging micro-engineering techniques such as micromachining, microfluidic and 3D printing techniques. In recent years, glomerular chips based on materials such as Polydimethylsiloxane (PDMS) and Plexiglass (PMMA) combined with extracellular matrices such as hydrogels and decellularized tissues have been widely reported. However, the basal membrane constructed by the existing glomerular chip is a flat-shaped rather than spherical twisted-shaped structure, resulting in limited glomerular filtration function. In summary, the current in vitro glomerular model is still low in degree of biomimetic.

Therefore, how to improve the bionics of glomerular chips is a technical problem to be solved by those skilled in the art.

Disclosure of Invention

The invention aims to provide a glomerular chip, which has improved bionics.

In order to achieve the above object, the present invention provides a glomerular chip comprising a first chip, a second chip, a third chip and a fourth chip stacked in this order from top to bottom;

the first chip is provided with a first filtrate outlet channel, a first arteriole outlet channel, an arteriole inlet channel and a first liquid accommodating channel communicated with the arteriole inlet channel, the opening of the first liquid accommodating channel is downward, and the first filtrate outlet channel, the first arteriole outlet channel and the arteriole inlet channel are not communicated with each other;

the second chip is provided with a liquid channel, a Bao cavity, a first filtrate through hole, a first outlet through hole, a first liquid accommodating groove and a liquid conveying through hole communicated with the Bao cavity, the first liquid accommodating groove is communicated with the first liquid accommodating channel, the liquid channel comprises an arteriole afferent micro-channel with an inlet connected with the outlet of the first liquid accommodating groove and a Bao cavity inlet channel communicated with the arteriole afferent micro-channel and the Bao cavity, the Bao cavity is used for accommodating hollow fibers, and two ends of the hollow fibers are respectively embedded into the Bao cavity inlet channel, so that a culture medium can only pass through the inner lumen of the hollow fibers;

the third chip is provided with a second liquid accommodating groove, an arteriole outgoing micro-channel, a second arteriole outlet channel, a Bao's through hole and a filtrate second through hole communicated with the filtrate first through hole, the arteriole outgoing micro-channel and the second arteriole outlet channel are both communicated with the second liquid accommodating groove, the Bao's through hole is communicated with the liquid conveying through hole, the arteriole outgoing micro-channel is communicated with the Bao's cavity inlet channel, the inlet of the arteriole outgoing micro-channel is connected with the outlet end through hole of the Bao's cavity inlet channel, and the Bao's through hole and the filtrate second through hole are not communicated with each other;

the fourth chip is provided with a third liquid containing groove of the near-end small tube micro-channel and a second filtrate outlet channel communicated with the third liquid containing groove, the third liquid containing groove and the Bao's through holes are communicated with the near-end small tube micro-channel, and the second filtrate outlet channel is communicated with the filtrate second through hole.

Preferably, the liquid conveying through holes, the baud cavities and the liquid channels are all multiple, the baud cavities and the liquid channels are in one-to-one correspondence, and the liquid conveying through holes and the baud cavities are in one-to-one correspondence.

Preferably, the plurality of the pall through holes and the plurality of the arteriole efferent micro-channels are all in one-to-one correspondence with the pall cavities, and the arteriole efferent micro-channels are in one-to-one correspondence with the pall cavity inlet channels.

Preferably, the number of the proximal small tube micro-channels is multiple, and the proximal small tube micro-channels are in one-to-one correspondence with the Bobs through holes.

Preferably, the first, second, third and fourth chips are fastened by threaded fasteners.

Preferably, the plurality of the abalone cavities are identical in structure, wherein the abalone cavities are respectively obtained by taking the first liquid accommodating groove as a preset angle from the previous abalone cavity.

Preferably, the width of the arteriole efferent micro-channel is 0.5 mm-1.5 mm, and the depth is 0.3 mm-1.0 mm;

and/or the width of the outlet channel of the second arteriole is 0.5 mm-1.5 mm and the depth is 0.3 mm-1.0 mm;

and/or the width of the micro-channel of the near-end small tube is 0.5 mm-1.5 mm, and the depth is 0.3 mm-1.0 mm;

and/or the width of the second filtrate outlet channel is 0.5 mm-1.5 mm and the depth is 0.3 mm-1.0 mm.

Preferably, the inlet end of the arteriole efferent microchannel is provided with a groove with a diameter larger than the width of the arteriole efferent microchannel.

Preferably, the outlet end of the proximal tubule microchannel is provided with a groove having a diameter larger than the width of the proximal tubule microchannel.

Preferably, the projection is from top to bottom:

the first liquid accommodating channel, the first liquid accommodating groove, the second liquid accommodating groove and the third liquid accommodating groove are overlapped;

and/or the through holes at the outlet end of the Bao cavity inlet channel are respectively overlapped with the circular grooves at the inlet end of the arteriole outgoing micro-channel;

and/or the liquid conveying through holes are respectively overlapped with the Bob through holes.

In the above technical solution, the glomerular chip provided by the present invention includes a first chip, a second chip, a third chip and a fourth chip stacked from top to bottom; the first chip is provided with a filtrate outlet channel, an arteriole inlet channel and a first liquid accommodating channel communicated with the arteriole inlet channel; the second chip is provided with a liquid channel, a Bao cavity, a first filtrate through hole, a first outlet through hole, a first liquid accommodating groove and a liquid conveying through hole communicated with the Bao cavity, the liquid accommodating groove is communicated with the first liquid accommodating channel, and the liquid channel comprises an arteriole afferent micro-channel with an inlet connected with an outlet of the liquid accommodating groove and a Bao cavity inlet channel communicated with the arteriole afferent micro-channel and the Bao cavity; the third chip is provided with a second liquid accommodating groove, an arteriole outgoing micro-channel, an arteriole outlet channel, a Bao's through hole and a filtrate second through hole communicated with the filtrate first through hole, wherein the arteriole outgoing micro-channel and the arteriole outlet channel are both communicated with the second liquid accommodating groove, the Bao's through hole is communicated with the liquid conveying through hole, and the arteriole outgoing micro-channel is communicated with the Bao's cavity inlet channel; the fourth chip is provided with a near-end small pipe micro-channel, a third liquid accommodating groove and a filtrate outlet channel communicated with the third liquid accommodating groove, the third liquid accommodating groove and the Bao's through hole are both communicated with the near-end small pipe micro-channel, and the filtrate outlet channel is communicated with the filtrate second through hole.

As can be seen from the above description, in the glomerular chip provided in the present application, by setting the bowing cavity, a functional area is provided for loading the glomerular capillary vessel into the bowing cavity on the chip, so as to form a bionic glomerular capillary vessel-like structure, and further improve the imitativeness of the glomerular chip.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments or the description of the prior art will be briefly described below, and it is obvious that the drawings in the following description are only embodiments of the present invention, and that other drawings can be obtained according to the provided drawings without inventive effort for a person skilled in the art.

FIG. 1 is an exploded view of a glomerular chip provided in an embodiment of the present invention;

fig. 2 is a schematic structural diagram of a first chip according to an embodiment of the present invention;

FIG. 3 is a schematic diagram of a second chip according to an embodiment of the present invention;

fig. 4 is a schematic structural diagram of a third chip according to an embodiment of the present invention;

fig. 5 is a schematic structural diagram of a fourth chip according to an embodiment of the present invention.

Wherein in fig. 1-5: 1-first chip, 2-second chip, 3-third chip, 4-fourth chip, 5-arteriole inlet channel, 6-first filtrate outlet channel, 7-first arteriole outlet channel, 8-first liquid receiving channel, 9-screw hole, 10-screw hole, 11-screw hole, 12-first liquid receiving groove, 13-arteriole afferent microchannel, 14-baud chamber inlet channel, 15-baud chamber, 16-baud chamber, 17-baud chamber, 18-baud chamber, 19-baud chamber, 20-baud chamber, 21-filtrate first through hole, 22-first outlet through hole, 23-liquid delivery through hole, 24-arteriole afferent microchannel, 25-arteriole afferent microchannel a 26-arteriole efferent microchannel, a 27-arteriole efferent microchannel, a 28-arteriole efferent microchannel, a 29-arteriole efferent microchannel, a 30-second arteriole outlet channel, a 31-baud through-hole, a 32-baud through-hole, a 33-baud through-hole, a 34-baud through-hole, a 35-baud through-hole, a 36-baud through-hole, a 37-second liquid receiving slot, a 38-filtrate second through-hole, a 39-proximal tubule microchannel, a 40-proximal tubule microchannel, a 41-proximal tubule microchannel, a 42-proximal tubule microchannel, a 43-proximal tubule microchannel, a 44-proximal tubule microchannel, a 45-second filtrate outlet channel, a 46-third liquid receiving slot.

Detailed Description

The core of the invention is to provide a glomerular chip with improved biomimetic properties.

The present invention will be described in further detail below with reference to the drawings and embodiments, so that those skilled in the art can better understand the technical solutions of the present invention.

Please refer to fig. 1 to 5.

In one embodiment, the glomerular chip provided in the embodiment of the present invention includes a first chip 1, a second chip 2, a third chip 3 and a fourth chip 4 stacked from top to bottom. The shapes of the first chip 1, the second chip 2, the third chip 3 and the fourth chip 4 can be round, round corner rectangular or rectangular, the areas of the upper surfaces and the lower surfaces of the first chip 1, the second chip 2, the third chip 3 and the fourth chip 4 are equal, and the specific areas can be 110mm and 90mm in length and width respectively. The thickness of the first chip 1 is not too thin, which is generally larger than 8mm, and can be 10mm specifically, so that screw holes can be conveniently drilled at the side edges of the chip, and the first chip 1 is too thin, so that leakage of culture medium can be easily caused in the chip operation process. The thickness of the second chip 2 and the third chip 3 is 2mm to 4mm, and is not preferably too thick or too thin. If the two layers of chips in the middle layer are too thick, the assembly of the whole glomerular chip is not facilitated; if the middle two-layer chip is too thin, it is easily bent and deformed during the assembly of the chip. The thickness of the fourth chip 4 is generally greater than 3mm, specifically may be 4mm, and when too thin, the fourth chip is easy to bend and deform and is easy to leak the culture medium in the operation process of the chip.

In this embodiment, the materials of the first chip 1, the second chip 2, the third chip 3 and the fourth chip 4 may be organic glass (PMMA) or Polydimethylsiloxane (PDMS); in this example, the types and sources of the organic glass and PDMS are not particularly limited, and the organic glass and PDMS, which are well known to those skilled in the art, may be used and commercially available; as in the embodiments of the present invention, plexiglas available from su-state and da plastics, inc, or PDMS polymer available from michigan midland corning, usa may be used.

The first chip 1, the second chip 2, the third chip 3 and the fourth chip 4 are fastened by threaded fasteners. The first chip 1, the second chip 2, the third chip 3 and the fourth chip 4 may also be fastened by means of a snap-fit. Specifically, the first chip 1 is provided with

screw holes

9, 10 and 11, the

screw holes

9, 10 and 11 are circumferentially arranged around the first chip 1, and can be ten, and the second chip 2, the third chip 3 and the fourth chip 4 are respectively provided with mounting holes corresponding to the

screw holes

9, 10 and 11 so as to be integrally fixed.

The first chip is provided with a first filtrate outlet channel 6, a first arteriole outlet channel 7, an arteriole inlet channel 5 and a first liquid containing channel 8 communicated with the arteriole inlet channel 5, the opening of the first liquid containing channel 8 is downward, and specifically, the arteriole inlet 5 is communicated with the first liquid containing channel 8 at the bottom end of the first layer chip 1. The arteriole inlet channel 5, the first filtrate outlet channel 6 and the first arteriole outlet channel 7 are independently arranged, i.e. are mutually disjoint, and are not communicated. Specifically, the arteriole inlet channel 5 is responsible for delivering culture medium and perfusing a cell suspension, the first filtrate outlet channel 6 is responsible for collecting culture medium that is not filtered by glomeruli, and the first arteriole outlet channel 7 is responsible for collecting filtrate after glomeruli filtration.

The second chip is provided with a liquid channel,

bob cavities

15, 16, 17, 18, 19 and 20, a first through hole 21 for filtrate, a first outlet through hole 22, a first liquid accommodating groove 12 and a liquid conveying through hole 23 communicated with the Bob cavities 15, 16, 17, 18, 19 and 20, the first liquid accommodating groove 12 is communicated with the first liquid accommodating channel 8, and the liquid channel comprises an arteriole afferent micro-channel 13 with an inlet connected with the outlet of the first liquid accommodating groove 12 and a Bob cavity inlet channel 14 communicated with the arteriole afferent micro-channel 13 and the Bob cavities 15, 16, 17, 18, 19 and 20. Specifically, the number of the

pall cavities

15, 16, 17, 18, 19, 20 may be one or at least two, and as shown, the number of the pall cavities is six, which are respectively the pall cavity 15, the pall cavity 16, the pall cavity 17, the pall cavity 18, the pall cavity 19 and the pall cavity 20.

When the number of the liquid delivery through holes 23, the

abalone cavities

15, 16, 17, 18, 19, 20 and the number of the liquid channels are all plural, the

abalone cavities

15, 16, 17, 18, 19, 20 are in one-to-one correspondence, the number of the liquid delivery through holes 23, the

abalone cavities

15, 16, 17, 18, 19, 20 are in one-to-one correspondence, and specifically, the number of the liquid delivery through holes 23, the arteriole afferent micro-channels 13 and the number of the abalone cavity inlet channels 14 are all equal to the number of the

abalone cavities

15, 16, 17, 18, 19, 20.

Specifically, the first liquid accommodating groove 12 is communicated with the bottom end opening of the first liquid accommodating channel 8 and receives the culture medium flowing in from the arteriole inlet channel 5 in the first chip 1; the first liquid accommodating groove 12 is communicated with the arteriole afferent micro-channel 13 and the Bao cavity inlet channel 14, and is used for conveying the culture medium to the Bao cavities 15, 16, 17, 18, 19 and 20; the

Bob chambers

15, 16, 17, 18, 19 and 20 are the main functional areas of the whole chip for loading of glomerular capillary clusters.

The plurality of

abalone chambers

15, 16, 17, 18, 19, 20 are the same in structure, wherein the

abalone chambers

15, 16, 17, 18, 19, 20 are respectively obtained by taking the first liquid accommodating groove 12 as a preset angle from the

previous abalone chambers

15, 16, 17, 18, 19, 20. Specifically, the diameter of the

abalone chambers

15, 16, 17, 18, 19 and 20 is 10mm, the depth is 1mm, and the

abalone chambers

16, 17, 18, 19 and 20 are respectively obtained by rotating 36 degrees around the first liquid accommodating groove 12 as the center of a circle in the previous abalone chamber. The chamber inlet passages 14 tangential to the

chambers

15, 16, 17, 18, 19, 20 are 3mm wide and the chamber inlet passages 14 have a depth of 1mm.

The third chip is provided with a second liquid accommodating groove 37, arteriole

outgoing micro-channels

24, 25, 26, 27, 28, 29, a second arteriole outlet channel 30, bob through

holes

31, 32, 33, 34, 35, 36 and a filtrate second through hole 38 communicated with the filtrate first through hole 21, the arteriole

outgoing micro-channels

24, 25, 26, 27, 28, 29 and the second arteriole outlet channel 30 are communicated with the second liquid accommodating groove 37, the Bob through

holes

31, 32, 33, 34, 35, 36 are communicated with the liquid conveying through hole 23, and the arteriole

outgoing micro-channels

24, 25, 26, 27, 28, 29 are communicated with the Bob cavity inlet channel 14. Preferably, the plurality of Bob's through

holes

31, 32, 33, 34, 35, 36 and arteriole efferent microchannels 24, 25, 26, 27, 28, 29 are provided, and the Bob's through

holes

31, 32, 33, 34, 35, 36 are in one-to-one correspondence with the Bob's

chambers

15, 16, 17, 18, 19, 20, and the arteriole efferent microchannels 24, 25, 26, 27, 28, 29 are in one-to-one correspondence with the Bob's chamber inlet channels 14.

Specifically, the arteriole efferent microchannels 24, 25, 26, 27, 28, 29 and the second arteriole outlet channel 30 on the third chip 3 are in communication with the second fluid reservoir 37; the baud's through

holes

31, 32, 33, 34, 35, 36 and the filtrate second through holes 38 are independently provided, and do not communicate with each other, i.e., do not intersect with each other. The inlet of arteriole efferent microchannels 24, 25, 26, 27, 28 and 29 are connected to the outlet port throughbore of the Bob's lumen inlet channel 14. Specifically, arteriole

efferent microchannels

24, 25, 26, 27, 28 and 29 collect medium from the second chip 2 that has not been glomerular filtered, pool on the second arteriole outlet channel 30 and exit through the first outlet through-hole in the second chip 2.

The arteriole efferent microchannels 24, 25, 26, 27, 28, 29 have a width of 0.5mm to 1.5mm, specifically 1mm, and a length of 28.5mm. The depth is 0.3mm to 1.0mm, and may be specifically 0.5mm.

The second arteriole outlet channel 30 has a length of 30mm, a width of 0.5mm to 1.5mm, specifically 1mm, and a depth of 0.3mm to 1.0mm, specifically 0.5mm.

The fourth chip is provided with proximal tubule micro-channels 39, 40, 41, 42, 43, 44, a third liquid containing groove 46 and a second filtrate outlet channel 45 communicated with the third liquid containing groove 46, wherein the third liquid containing groove 46 and the Bob through

holes

31, 32, 33, 34, 35, 36 are communicated with the proximal tubule micro-channels 39, 40, 41, 42, 43, 44, and the second filtrate outlet channel 45 is communicated with the filtrate second through hole 38.

The proximal tubule microchannels 39, 40, 41, 42, 43, 44 and the second filtrate outlet channel 45 are in communication with a third liquid reservoir 46. Specifically, the proximal tubule microchannels 39, 40, 41, 42, 43, 44 are 41.5mm long, the proximal tubule microchannels 39, 40, 41, 42, 43, 44 have a width of 0.5mm to 1.5mm, specifically 1mm, a depth of 0.3mm to 1.0mm, specifically 0.5mm, and the second filtrate outlet channel 45 is 30mm long, 0.5mm to 1.5mm wide, specifically 1mm, and a depth of 0.3mm to 1.0mm, specifically 0.5mm. The glomerulfiltered medium is collected via proximal tubule microchannels 39, 40, 41, 42, 43, 44 onto a second filtrate outlet channel 45 and finally returned to the first chip 1 via filtrate second through holes 38 on the third chip 3 and filtrate first through holes 21 on the second chip 2.

As shown, the number of proximal tubule micro-channels 39, 40, 41, 42, 43, 44 is plural, and the proximal tubule micro-channels 39, 40, 41, 42, 43, 44 are in one-to-one correspondence with the bowden through

holes

31, 32, 33, 34, 35, 36.

The inlet ends of the arteriole efferent microchannels 24, 25, 26, 27, 28, 29 are provided with grooves having a diameter greater than the width of the arteriole efferent microchannels 24, 25, 26, 27, 28, 29.

The outlet ends of the proximal tubule microchannels 39, 40, 41, 42, 43, 44 are provided with grooves having a diameter larger than the width of the proximal tubule microchannels 39, 40, 41, 42, 43, 44.

Based on the schemes, the projection is from top to bottom:

the first liquid containing channel 8, the first liquid containing groove 12, the second liquid containing groove 37, and the third liquid containing groove 46 are overlapped.

The outlet end through holes of the Bao cavity inlet channel 14 are respectively overlapped with the round grooves of the inlet ends of the arteriole

outgoing micro-channels

24, 25, 26, 27, 28 and 29.

The liquid transport through-holes 23 coincide with the Bobs through-

holes

31, 32, 33, 34, 35, 36, respectively.

The assembly method of the glomerular chip design can be as follows: the first chip 1, the second chip 2, the third chip 3 and the fourth chip 4 are arranged in sequence from top to bottom. The first liquid containing channel 8 at the bottom of the first chip 1 is overlapped with the first liquid containing groove at the top of the second chip 2; overlapping the small hole at one end of the small artery outlet channel on the first chip 1 with the first outlet through hole 22 on the second chip 2; overlapping the hole of the first filtrate outlet channel 6 at the bottom of the first chip 1 with the first filtrate through hole 21 on the second chip 2; the relative positions of the first chip 1 and the second chip 2 can be determined by the alignment of these structures. Then, the tail end through holes of the Bao cavity inlet channel 14 on the second chip 2 are respectively overlapped with the front end circular grooves of the arteriole

outgoing micro-channels

24, 25, 26, 27, 28 and 29 on the third chip 3; overlapping the filtrate first through hole 21 on the second chip 2 with the filtrate second through hole 38 on the third chip 3; overlapping the first outlet through hole 22 on the second chip 2 with the end circular hole of the second arteriole outlet channel 30 on the third chip 3; the relative positions of the second chip 2 and the third chip 3 can be determined by the alignment of these structures. Then, the second liquid-receiving groove 37 on the third chip 3 is aligned with the third liquid-receiving groove 46 on the fourth chip 4, and the Bobs's through-

holes

31, 32, 33, 34, 35, 36 on the third chip 3 are aligned with the inlet circular grooves of the proximal tubule micro-channels 44, 43, 42, 41, 40, 39 on the fourth chip 4, respectively; aligning the second filtrate through hole 38 of the third chip 3 with the circular groove of the outlet end of the second filtrate outlet channel 45 of the fourth chip 4; by alignment of these structures, the relative positions of the third chip 3 and the fourth chip 4 can be determined and the assembly of the glomerular chip design can be completed. Specifically, the first chip 1, the second chip 2, the third chip 3 and the fourth chip 4 may be fastened by edge 10 screws. Specifically, between the first chip 1 and the second chip 2, between the second chip 2 and the third chip 3, between the third chip 3 and the fourth chip 4, a silica gel film can be respectively sandwiched between the first chip and the second chip, so that the sealing performance of the chips is improved.

The glomerular chip provided by the application is designed for in vitro construction of a glomerular model, and the specific process is as follows:

wrapping the hollow fiber end (-3 cm) of the plasma separator with equal amount of silica gel, and drying in a 60 deg.C drying oven for 20min until the silica gel is completely dried; cutting the dried hollow fiber along the middle of the silica gel wrapping part to expose a hollow channel, and bending the hollow fiber into an 'oc' shape; placing the bent hollow fiber in the Bob's chambers 15, 16, 17, 18, 19, 20 on the second chip 2, specifically placing the main part of the bent hollow fiber in the Bob's chambers 15, 16, 17, 18, 19, 20, and embedding the two ends of the hollow fiber in the second chip 2The abalone cavity inlet channel 14 is fixed by silica gel, so that the culture medium can only pass through the inner lumen of the hollow fiber, thereby simulating the capillary cluster structure of human glomeruli; the hollow fibers were coated with human fibronectin (0.5 mg/mL; formulated directly with sterile PBS) for 1 hour (37 ℃,5% CO) ₂ ) To support podocyte adhesion and growth; mu.L of podocyte suspension (1X 10) ⁷ cells/mL) was inoculated to the hollow fiber surfaces in the bowden cavities 15, 16, 17, 18, 19 and 20, and incubated in an incubator for two hours to promote adhesion of cells on the hollow fiber tube surfaces. A silicone film (thickness: -0.5 mm) was sandwiched between each two layers of chips, and the chip assembly process was completed. 1mL of endothelial cell suspension (2X 10) was injected using a 5mL syringe ⁵ cells/mL) was infused into the hollow fiber through arteriole inlet 5 on the first chip 1 and allowed to stand for 2h to support adhesion of endothelial cells on the inner surface of the hollow fiber. Sterile peristaltic tubing was connected to a flask of medium containing 60mL of cell culture medium and placed in an incubator. The culture medium was driven by peristaltic pump at 0.2mL/h through the arteriole inlet 5 on the first chip 1, then into the capillaries in the six

bowden cavities

15, 16, 17, 18, 19 and 20, respectively, through the arteriole efferent microchannels 24, 25, 26, 27, 28 and 29 on the third chip 3, and finally back into the culture medium bottle from the arteriole outlet 7 on the first chip 1, and the other portion of the culture medium was filtered by the capillaries (glomerular capillary structures) in the bowden cavities 15, 16, 17, 18, 19 and 20, then through the proximal tubule microchannels 39, 40, 41, 42, 43, 44 on the fourth chip 4 and back into the filtrate outlet 6 on the first chip 1, simulating blood filtration behavior in humans.

According to the glomerular capillary blood vessel simulation method, simulation of an external glomerular capillary blood vessel structure can be achieved, meanwhile, separated capillary blood vessel clusters and abalone cavities can be achieved, a solution idea is provided for the problem that the glomerular model existing at present is low in bionic degree, based on the glomerular chip, a hollow fiber bundle is packaged in the abalone cavities of the chip in an 'oc' shape mode, foot cells and endothelial cells are inoculated to two sides of a fiber tube respectively, and therefore the glomerular capillary blood vessel structure is simulated in vitro, the separated glomerular capillary blood vessel clusters and the abalone cavities are achieved, and physiological similarity of the glomerular model is improved. The filtrate outlet 6 on the chip 1 simulates the blood filtration behaviour in a human body.

In the present specification, each embodiment is described in a progressive manner, and each embodiment is mainly described in a different point from other embodiments, and identical and similar parts between the embodiments are all enough to refer to each other.

The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. The glomerular chip is characterized by comprising a first chip (1), a second chip (2), a third chip (3) and a fourth chip (4) which are sequentially overlapped from top to bottom;

the first chip (1) is provided with a first filtrate outlet channel (6), a first arteriole outlet channel (7), an arteriole inlet channel (5) and a first liquid accommodating channel (8) communicated with the arteriole inlet channel (5), the opening of the first liquid accommodating channel (8) is downward, and the first filtrate outlet channel (6), the first arteriole outlet channel (7) and the arteriole inlet channel (5) are not communicated with each other;

the second chip (2) is provided with a liquid channel, a Bao's cavity (15), (16), (17), (18), (19), (20), a first through hole (21) for filtrate, a first outlet through hole (22), a first liquid accommodating groove (12) and a liquid conveying through hole (23) communicated with the Bao's cavity (15), (16), (17), (18), (19), (20), the first liquid accommodating groove (12) is communicated with the first liquid accommodating channel (8), the liquid channel comprises an arteriole afferent micro-channel (13) with an inlet connected with the outlet of the first liquid accommodating groove (12) and a Bao's cavity inlet channel (14) communicated with the arteriole afferent micro-channel (13) and the Bao's cavities (15), (16), (17), (18), (19), (20), the Bao's cavities (15), (16), (17), (18), (19), (20) are used for accommodating hollow fibers, and two ends of the hollow fibers are respectively embedded into the Bao's cavity inlet channel (14) so that culture mediums can only pass through the inner tube cavities of the hollow fibers;

the third chip (3) is provided with a second liquid accommodating groove (37), an arteriole outgoing micro-channel (24), (25), (26), (27), (28), (29), a second arteriole outlet channel (30), a Bao's through-hole (31), (32), (33), (34), (35), (36) and a filtrate second through-hole (38) communicated with the filtrate first through-hole (21), the arteriole outgoing micro-channel (24), (25), (26), (27), (28), (29) and the second arteriole outlet channel (30) are communicated with the second liquid accommodating groove (37), the Bao's through-hole (31), (32), (33), (34), (35), (36) are communicated with the liquid conveying through-hole (23), the arteriole outgoing micro-channel (24), (25), (26), (27), (28), (29) are communicated with the Bao's cavity inlet channel (14), the inlets of the arteriole outgoing micro-channel (24), (25), (26), (27), (28), (29) are connected with the outlet end of the Bao's cavity inlet channel (14), the Bob through holes (31), (32), (33), (34), (35), (36) and the filtrate second through holes (38) are not communicated with each other;

the fourth chip (4) is provided with proximal small tube micro-channels (39), (40), (41), (42), (43), (44), a third liquid accommodating groove (46) and a second filtrate outlet channel (45) communicated with the third liquid accommodating groove (46), the third liquid accommodating groove (46) and the Bob through holes (31), (32), (33), (34), (35), (36) are communicated with the proximal small tube micro-channels (39), (40), (41), (42), (43), (44), and the second filtrate outlet channel (45) is communicated with the filtrate second through hole (38).

2. The glomerular chip of claim 1, wherein the liquid delivery through-hole (23), the bowden cavities (15), (16), (17), (18), (19), (20) and the liquid channel are all plural, the bowden cavities (15), (16), (17), (18), (19), (20) and the liquid channel are in one-to-one correspondence, and the liquid delivery through-hole (23) and the bowden cavities (15), (16), (17), (18), (19), (20) are in one-to-one correspondence.

3. The glomerular chip of claim 2, wherein the bowden through holes (31), (32), (33), (34), (35), (36) and the arteriole efferent microchannels (24), (25), (26), (27), (28), (29) are all plural, and the bowden through holes (31), (32), (33), (34), (35), (36) are in one-to-one correspondence with the bowden lumens (15), (16), (17), (18), (19), (20), the arteriole efferent microchannels (24), (25), (26), (27), (28), (29) are in one-to-one correspondence with the bowden lumen inlet channels (14).

4. The glomerular chip of claim 3, wherein the proximal tubule micro-channels (39), (40), (41), (42), (43), (44) are plural, and the proximal tubule micro-channels (39), (40), (41), (42), (43), (44) are in one-to-one correspondence with the bowden through holes (31), (32), (33), (34), (35), (36).

5. The glomerular chip of claim 1, wherein the first, second, third, and fourth chips are secured by threaded fasteners.

6. The glomerular chip of claim 1, wherein the plurality of bowden cavities (15), (16), (17), (18), (19), (20) are identical in structure, and wherein the bowden cavities (15), (16), (17), (18), (19), (20) are respectively obtained from the previous bowden cavity (15), (16), (17), (18), (19), (20) by taking the first liquid accommodating groove (12) as a preset angle.

7. The glomerular chip of claim 1, wherein the arteriole efferent microchannels (24), (25), (26), (27), (28), (29) have a width of 0.5mm to 1.5mm and a depth of 0.3mm to 1.0mm;

and/or the width of the second arteriole outlet channel (30) is 0.5mm to 1.5mm and the depth is 0.3mm to 1.0mm;

and/or the width of the proximal tubule micro-channels (39), (40), (41), (42), (43), (44) is 0.5 mm-1.5 mm and the depth is 0.3 mm-1.0 mm;

and/or the second filtrate outlet channel (45) has a width of 0.5mm to 1.5mm and a depth of 0.3mm to 1.0mm.

8. The glomerular chip of claim 1, wherein the inlet ends of the arteriole efferent microchannels (24), (25), (26), (27), (28), (29) are provided with grooves having a diameter greater than the width of the arteriole efferent microchannels (24), (25), (26), (27), (28), (29).

9. The glomerular chip of claim 1, wherein the outlet ends of the proximal tubule microchannels (39), (40), (41), (42), (43), (44) are provided with grooves having a diameter larger than the width of the proximal tubule microchannels (39), (40), (41), (42), (43), (44).

10. The glomerular chip of any one of claims 1-9, wherein the projections are from top to bottom:

the first liquid accommodating channel (8), the first liquid accommodating groove (12), the second liquid accommodating groove (37) and the third liquid accommodating groove (46) are overlapped;

and/or the through holes at the outlet end of the Bao cavity inlet channel (14) are respectively overlapped with the circular grooves at the inlet ends of the arteriole outgoing micro-channels (24), (25), (26), (27), (28) and (29);

and/or the liquid delivery through-hole (23) coincides with the Bob through-holes (31), (32), (33), (34), (35), (36), respectively.