CN113621515A

CN113621515A - Glomerulus chip

Info

Publication number: CN113621515A
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: 2021-11-09
Anticipated expiration: 2041-08-11
Also published as: CN113621515B

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, can realize the simulation of an in vitro glomerular capillary structure, can realize a separated capillary tube cluster and a Bob's cavity at the same time, and provides a solution for the problem of low bionic degree of a currently existing glomerular model.

Description

Glomerulus chip

Technical Field

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

Background

The kidneys are important organs of the human body and have an extremely complex structure and microenvironment. The kidney consists of millions of nephrons, of which glomeruli are important components. The glomerulus is responsible for performing the blood filtering function, and the filtering membrane of the glomerulus is a three-layer structure with a basement membrane in the middle. Glomerular filtration is the initial process of urine formation, and as blood flows through the glomeruli, most of the material is filtered by the glomerular capillaries, entering the lumen of bauer, and forming the crude urine. Studies have shown that most kidney diseases (e.g., chronic kidney disease, diabetic nephropathy, and nephrotic syndrome) are caused by glomerular dysfunction. Therefore, researchers have focused on building in vitro glomerular models to reveal the mechanisms of glomerular function.

Currently, there are two main types of glomerular models in vitro: kidney organoids and glomerular chips. The glomerulus constructed by the kidney organoid can express podocyte-associated protein and has the glomerulus filtering function. However, the existing kidney organoids have not realized a spherical glomerular structure with a bowden's lumen, and lack perfusable vascular networks, so that the application thereof is limited. Glomerular chips are in vitro glomerular models built based on emerging microengineering technologies (e.g., microfabrication, microfluidics, and 3D printing technologies). In recent years, a glomerular chip based on materials such as Polydimethylsiloxane (PDMS) and organic glass (PMMA) and combining an extracellular matrix such as hydrogel and acellular tissue has been widely reported. However, the basement membrane constructed by the existing glomerular chip is a flat structure rather than a spherical twisted structure, so that the glomerular filtration function is limited. In conclusion, the current in vitro glomerular models are still less biomimetic.

Therefore, how to improve the bionic property of the glomerular chip is a technical problem to be solved urgently by those skilled in the art.

Disclosure of Invention

The object of the present invention is to provide a glomerular chip having 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 which are first stacked from top to bottom;

the first chip is provided with a filtrate outlet channel, an arteriolar inlet channel and a first liquid containing channel communicated with the arteriolar inlet channel, the first liquid containing channel is downward opened, and the filtrate outlet channel, the arteriolar outlet channel and the arteriolar inlet channel are not communicated with each other;

the second chip is provided with a liquid channel, an Bausch cavity, a first filtrate through hole, a first outlet through hole, a first liquid containing groove and a liquid conveying through hole communicated with the Bausch cavity, the liquid containing groove is communicated with the first liquid containing groove, and the liquid channel comprises a arteriolar afferent microchannel with an inlet connected with an outlet of the liquid containing groove and a Bausch cavity inlet channel communicated with the arteriolar afferent microchannel and the Bausch cavity;

the third chip is provided with a second liquid containing groove, an arteriolar efferent micro-channel, an arteriolar outlet channel, a Bowden through hole and a filtrate second through hole communicated with the filtrate first through hole, the arteriolar efferent micro-channel and the arteriolar outlet channel are both communicated with the second liquid containing groove, the Bowden through hole is communicated with the liquid conveying through hole, the arteriolar efferent micro-channel is communicated with the Bowden cavity inlet channel, and the Bowden through hole and the filtrate second through hole are not communicated with each other;

the fourth chip is provided with a proximal tubule microchannel, a third liquid containing groove and a filtrate outlet channel communicated with the third liquid containing groove, the third liquid containing groove and the Babylonia through hole are communicated with the proximal tubule microchannel, and the filtrate outlet channel is communicated with the filtrate second through hole.

Preferably, the liquid conveying through holes, the Bob cavities and the liquid channels are all multiple, the Bob cavities correspond to the liquid channels one by one, and the liquid conveying through holes correspond to the Bob cavities one by one.

Preferably, the number of the Bob through holes and the number of the arteriolar efferent microchannels are both multiple, the Bob through holes correspond to the Bob cavities one by one, and the arteriolar efferent microchannels correspond to the Bob cavity inlet channels one by one.

Preferably, the number of the proximal tubule microchannels is multiple, and the proximal tubule microchannels correspond to the pall-lug through holes one by one.

Preferably, the first chip, the second chip, the third chip and the fourth chip are fastened by a threaded fastener.

Preferably, the plurality of the baud cavities have the same structure, and the previous baud cavities are respectively formed by a preset angle of the first liquid containing groove.

Preferably, the width of the arteriolar efferent microchannel is 0.5mm to 1.5mm, and the depth is 0.3mm to 1.0 m;

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

and/or the width of the proximal tubule microchannel is 0.5 mm-1.5 mm, and the depth is 0.3 mm-1.0 mm;

and/or the width of the 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 arteriolar efferent microchannel is provided with a groove having a diameter greater than the width of the arteriolar efferent microchannel.

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

Preferably, projecting from top to bottom:

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

and/or the through hole at the outlet end of the Babys cavity inlet channel is superposed with the circular groove at the inlet end of the arteriole efferent microchannel;

and/or the liquid conveying through holes are respectively superposed with the Babyloni 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 which are first stacked from top to bottom; the first chip is provided with a filtrate outlet channel, an arteriolar inlet channel and a first liquid containing channel communicated with the arteriolar inlet channel; the second chip is provided with a liquid channel, a Bausch cavity, a first filtrate through hole, a first outlet through hole, a first liquid containing groove and a liquid conveying through hole communicated with the Bausch cavity, the liquid containing groove is communicated with the first liquid containing groove, and the liquid channel comprises a arteriolar transfer micro channel with an inlet connected with an outlet of the liquid containing groove and a Bausch cavity inlet channel communicated with the arteriolar transfer micro channel and the Bausch cavity; the third chip is provided with a second liquid containing groove, an arteriolar efferent micro-channel, an arteriolar outlet channel, a Boehringer through hole and a filtrate second through hole communicated with the filtrate first through hole, the arteriolar efferent micro-channel and the arteriolar outlet channel are both communicated with the second liquid containing groove, the Boehringer through hole is communicated with the liquid conveying through hole, and the arteriolar efferent micro-channel is communicated with the Boehringer cavity inlet channel; the fourth chip is provided with a near-end tubule microchannel, a third liquid containing groove and a filtrate outlet channel communicated with the third liquid containing groove, the third liquid containing groove and the Babyloni through hole are communicated with the near-end tubule microchannel, and the filtrate outlet channel is communicated with the filtrate second through hole.

According to the description, in the glomerular chip provided by the application, the Bausch cavities are arranged, so that the functional regions are provided for loading glomerular capillaries into the Bausch cavities on the chip, the bionic glomerular capillary-like structure is formed, and the bionic property of the glomerular chip is improved.

Drawings

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

FIG. 1 is an exploded view of a glomerular chip provided by 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 structural 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 FIGS. 1-5: 1-first chip, 2-second chip, 3-third chip, 4-fourth chip, 5-arteriole inlet channel, 6-filtrate outlet channel, 7-arteriole outlet channel, 8-liquid first liquid holding channel, 9-screw hole, 10-screw hole, 11-screw hole, 12-first liquid holding tank, 13-arteriole afferent microchannel, 14-Babys cavity inlet channel, 15-Babys cavity, 16-Babys cavity, 17-Babys cavity, 18-Babys cavity, 19-Babys cavity, 20-Babys cavity, 21-filtrate first through hole, 22-first outlet through hole, 23-liquid delivery through hole, 24-arteriole efferent microchannel, 25-arteriole efferent microchannel, 26-arteriole efferent microchannel, 27-arteriolar efferent microchannel, 28-arteriolar efferent microchannel, 29-arteriolar efferent microchannel, 30-arteriolar outlet channel, 31-baumannii through hole, 32-baumannii through hole, 33-baumannii through hole, 34-baumannii through hole, 35-baumannii through hole, 36-baumannii through hole, 37-second liquid containing tank, 38-filtrate second through hole, 39-proximal tubule microchannel, 40-proximal tubule microchannel, 41-proximal tubule microchannel, 42-proximal tubule microchannel, 43-proximal tubule microchannel, 44-proximal tubule microchannel, 45-filtrate outlet channel, 46-third liquid containing tank.

Detailed Description

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

In order to make the technical solutions of the present invention better understood by those skilled in the art, the present invention will be further described in detail with reference to the accompanying drawings and embodiments.

Please refer to fig. 1 to 5.

In one embodiment, the glomerular chip provided by the embodiment of the 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 may be circular, rectangular with round corners or rectangular, the areas of the upper surface and the lower surface of the first chip 1, the second chip 2, the third chip 3 and the fourth chip 4 are equal, and the specific areas may be 110mm and 90mm respectively for length and width. The thickness of the first chip 1 is not suitable to be too thin, generally larger than 8mm, and specifically can be 10mm, so that a screw hole can be conveniently drilled at the side edge of the chip, and the culture medium is easy to leak in the operation process of the chip because the first chip 1 is too thin. The thickness of the second chip 2 and the third chip 3 is 2mm to 4mm, and the thickness is not too thick or too thin. If the two layers of the chip at the middle layer are too thick, the assembly of the whole glomerular chip is not facilitated; if the two intermediate layers of chips are too thin, they may be easily bent and deformed during the assembly of the chips. The thickness of the fourth chip 4 is generally greater than 3mm, and specifically may be 4mm, and too thin may easily bend and deform and may easily cause leakage of the culture medium during operation of the chip.

In this embodiment, the material 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 the present embodiment, there is no particular limitation on the types and sources of the organic glass and PDMS, and the organic glass and PDMS well known to those skilled in the art may be used and may be obtained from the market; as in the embodiments of the present invention, plexiglass available from Suzhou Anhe Dai plastics, Inc. or PDMS polymers available from Midland Corning, Michigan, USA may be used.

The first chip 1, the second chip 2, the third chip 3, and the fourth chip 4 are fastened by a threaded fastener. The first chip 1, the second chip 2, the third chip 3 and the fourth chip 4 may also be fastened by snaps. Specifically, be equipped with

screw hole

9, 10, 11 on first chip 1,

screw hole

9, 10, 11 are around 1 circumference setting of first chip, specifically can be ten, and second chip 2, third chip 3 and fourth chip 4 are equallyd divide and are do not be equipped with the mounting hole that corresponds the setting with

screw hole

9, 10, 11 to monolithic stationary.

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

The second chip is provided with a liquid channel, Bausch cavities 15, 16, 17, 18, 19 and 20, a first filtrate through hole 21, a first outlet through hole 22, a first liquid containing groove 12 and a liquid conveying through hole 23 communicated with the Bausch cavities 15, 16, 17, 18, 19 and 20, wherein the liquid containing groove 12 is communicated with the first liquid containing channel 8, and the liquid channel comprises a arteriolar transfer microchannel 13 with an inlet connected with an outlet of the liquid containing groove 12 and a Bausch cavity inlet channel 14 communicated with the arteriolar transfer microchannel 13 and the Bausch cavities 15, 16, 17, 18, 19 and 20. Specifically, there may be one or at least two of the

baud cavities

15, 16, 17, 18, 19, and 20, and as shown, there are six baud cavities, which are the baud cavity 15, the baud cavity 16, the baud cavity 17, the baud cavity 18, the baud cavity 19, and the baud cavity 20.

When the liquid conveying through holes 23, the bauer cavities 15, 16, 17, 18, 19, 20 and the liquid channels are all multiple, the bauer cavities 15, 16, 17, 18, 19, 20 and the liquid channels are in one-to-one correspondence, the liquid conveying through holes 23 and the bauer cavities 15, 16, 17, 18, 19, 20 are in one-to-one correspondence, and specifically, the liquid conveying through holes 23, the arteriole afferent micro-channel 13 and the bauer cavity inlet channel 14 are equal in number to the bauer cavities 15, 16, 17, 18, 19, 20.

Specifically, the first liquid containing groove 12 is communicated with the bottom opening of the first liquid containing channel 8 and receives the culture medium flowing from the arteriolar inlet channel 5 in the first chip 1; the first liquid containing groove 12 is communicated with the arteriolar afferent microchannel 13 and the Babyloni cavity inlet channel 14, and conveys the culture medium to the Babyloni cavities 15, 16, 17, 18, 19 and 20; the

Bowden chambers

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

The plurality of the

baud cavities

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

baud cavities

15, 16, 17, 18, 19, 20 are respectively obtained by the

previous baud cavity

15, 16, 17, 18, 19, 20 by using the first liquid containing groove 12 as a preset angle. Specifically, the diameters of the bauer cavities 15, 16, 17, 18, 19, and 20 are 10mm, the depths thereof are 1mm, and the bauer cavities 16, 17, 18, 19, and 20 are respectively obtained by rotating the previous bauer cavity by 36 degrees around the first liquid containing groove 12. The width of the Bausch cavity inlet channel 14 tangent to the Bausch cavities 15, 16, 17, 18, 19 and 20 is 3mm, and the depth of the fluid channel of the Bausch cavity inlet channel 14 is 1 mm.

The third chip is provided with a second liquid containing groove 37, arteriolar efferent micro-channels 24, 25, 26, 27, 28, 29, an arteriolar outlet channel 30, Bowden through

holes

31, 32, 33, 34, 35, 36 and a filtrate second through hole 38 communicated with the filtrate first through hole 21, the arteriolar efferent micro-channels 24, 25, 26, 27, 28, 29 and the arteriolar outlet channel 30 are communicated with the second liquid containing groove 37, the Bowden through

holes

31, 32, 33, 34, 35, 36 are communicated with the liquid conveying through hole 23, and the arteriolar efferent micro-channels 24, 25, 26, 27, 28, 29 are communicated with the Bowden cavity inlet channel 14. Preferably, the number of the bowden through

holes

31, 32, 33, 34, 35, 36 and the arteriolar

efferent microchannels

24, 25, 26, 27, 28, 29 is plural, and the bowden through

holes

31, 32, 33, 34, 35, 36 correspond to the bowden cavities 15, 16, 17, 18, 19, 20 one by one, and the arteriolar

efferent microchannels

24, 25, 26, 27, 28, 29 correspond to the bowden cavity inlet channels 14 one by one.

Specifically, the arteriolar

efferent microchannels

24, 25, 26, 27, 28, 29 and the arteriolar outlet channels 30 on the third chip 3 are all communicated with the second liquid containing groove 37; the Bob through

holes

31, 32, 33, 34, 35 and 36 and the second filtrate through hole 38 are independently arranged and are not communicated with each other, namely are not intersected with each other. The inlets of the arteriolar

efferent microchannels

24, 25, 26, 27, 28 and 29 are connected to the outlet end through holes of the bowden's lumen inlet channel 14. Specifically, the arteriolar

efferent microchannels

24, 25, 26, 27, 28 and 29 collect media from the second chip 2 that has not been filtered by the glomeruli, converge on the arteriolar exit channel 30 and exit through the first exit opening in the second chip 2.

The arteriolar

efferent microchannels

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

The arteriole outlet channel 30 is 30mm long, 0.5mm to 1.5m wide, specifically 1mm deep, 0.3mm to 1.0mm deep, specifically 0.5 mm.

The fourth chip has proximal tubule microchannels 39, 40, 41, 42, 43, 44, a third liquid receiving channel 46 and a filtrate outlet channel 45 communicating with the third liquid receiving channel 46, the third liquid receiving channel 46 and the bauschin's through

holes

31, 32, 33, 34, 35, 36 are all in communication with the proximal tubule microchannels 39, 40, 41, 42, 43, 44, and the filtrate outlet channel 45 is in communication with the filtrate second through hole 38.

The proximal tubule microchannels 39, 40, 41, 42, 43, 44 and filtrate outlet passage 45 are all in communication with a third liquid receiving 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, the 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 0.5 mm. The glomerular filtered medium collects through the proximal tubule microchannels 39, 40, 41, 42, 43, 44 onto the filtrate outlet channel 45 and eventually returns to the first chip 1 through the filtrate second through-hole 38 in the third chip 3 and the filtrate first through-hole 21 in the second chip 2.

As shown, the proximal tubule microchannels 39, 40, 41, 42, 43, 44 are plural, and the proximal tubule microchannels 39, 40, 41, 42, 43, 44 correspond to the

bowden cables

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

The inlet ends of the arteriolar

efferent microchannels

24, 25, 26, 27, 28, 29 are provided with grooves having a diameter greater than the width of the arteriolar

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 greater than the width of the proximal tubule microchannels 39, 40, 41, 42, 43, 44.

On the basis of the above schemes, projection is performed from top to bottom:

the liquid first liquid accommodating passage 8, the first liquid accommodating groove 12, the second liquid accommodating groove 37 and the third liquid accommodating groove 46 coincide.

The outlet end through holes of the Babys cavity inlet channel 14 are respectively overlapped with the circular grooves of the inlet ends of the arteriolar

efferent microchannels

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

The liquid-conveying through-holes 23 coincide with the bauer through-

holes

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

The assembly method of the glomerular chip design may be: 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 liquid first liquid containing channel 8 at the bottom of the first chip 1 is superposed with the first liquid containing groove at the top of the second chip 2; making the small hole at one end of the arteriole outlet channel on the first chip 1 coincide with the first outlet through hole 22 on the second chip 2; the hole of the filtrate outlet channel 6 at the bottom of the first chip 1 is superposed with the first through hole 21 of the filtrate on the second chip 2; the relative position of the first chip 1 and the second chip 2 can be determined by the alignment of these structures. Then, the through holes at the tail ends of the Bausch cavity inlet channels 14 on the second chip 2 are respectively superposed with the circular grooves at the front ends of the arteriolar

efferent microchannels

24, 25, 26, 27, 28 and 29 on the third chip 3; the first through hole 21 of the filtrate on the second chip 2 is superposed with the second through hole 38 of the filtrate on the third chip 3; the first outlet through hole 22 on the second chip 2 is superposed with the end round hole of the arteriole outlet channel 30 on the third chip 3; the relative position of the second chip 2 and the third chip 3 can be determined by the alignment of these structures. Then, the second liquid containing grooves 37 on the third chip 3 are aligned with the third liquid containing grooves 46 on the fourth chip 4, and the bauer through

holes

31, 32, 33, 34, 35, 36 on the third chip 3 are aligned with the inlet circular grooves of the proximal tubule microchannels 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 at the outlet end of the filtrate outlet channel 45 of the fourth chip 4; the relative positions of the third chip 3 and the fourth chip 4 can be determined by the alignment of these structures 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 10 screws at the edge. Specifically, between first chip 1 and second chip 2, between second chip 2 and third chip 3, can equally divide respectively through pressing from both sides a layer of pellosil between third chip 3 and the fourth chip 4, improve the leakproofness of chip.

The glomerular chip design that this application provided is used for external glomerular model of establishing, and specific process is:

wrapping the tail end (about 3cm) of the hollow fiber from the plasma separator with equal amount of silica gel, and drying in a drying oven at 60 deg.C 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 the hollow channel, and bending the hollow fiber into a shape of '. alpha'; placing the bent hollow fibers in the Bob cavities 15, 16, 17, 18, 19 and 20 on the second chip 2, specifically, placing the bent main parts of the hollow fibers in the Bob cavities 15, 16, 17, 18, 19 and 20, respectively embedding the two ends of the bent main parts into the Bob cavity inlet channels 14 on the Bob chip 2 on the second chip 2, and fixing the bent main parts by silica gel to ensure that the culture medium can only pass through the inner tube cavities of the hollow fibers, thereby simulating the human glomerular capillary cluster structure; the hollow fibers were coated with human fibronectin (0.5 mg/mL; prepared directly in sterile PBS) for 1 hour (37 ℃, 5% CO)₂) To support adhesion and growth of podocytes; 30 μ L of podocyte suspension (1X 10)⁷cells/mL) were seeded onto the hollow fiber surfaces in the cavities 15, 16, 17, 18, 19 and 20 of bowden and incubated in an incubator for two hours to promote cell adhesion 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 with a 5mL syringe⁵cells/mL) was perfused into the hollow fiber through the arteriole inlet 5 on the first chip 1 and left for 2h to support endothelial cell adhesion on the inner surface of the hollow fiber. The sterile peristaltic tubing was connected to a medium bottle containing 60mL of cell culture medium and placed in an incubator. The culture medium is driven by a peristaltic pump at a speed of 0.2mL/h, passes through the arteriole inlet 5 on the first chip 1, then enters the fiber tubes in the six

Bowden cavities

15, 16, 17, 18, 19 and 20, respectively, flows through the arteriole efferent microchannels 24, 25, 26, 27, 28 and 29 on the third chip 3, finally returns to the culture medium bottle from the arteriole outlet 7 on the first chip 1, and another part of the culture medium flows through the proximal tubule microchannel on the fourth chip 4 after being filtered by the fiber tubes (glomerular capillary structures) in the Bowden cavities 15, 16, 17, 18, 19 and

20The channels

39, 40, 41, 42, 43, 44 and back to the filtrate outlet 6 on the first chip 1 simulate the blood filtration behaviour in humans.

The simulation of in vitro glomerular capillary structure can be realized to this application, can realize the capillary blood vessel cluster and the bau chamber of separation simultaneously, provide the solution thinking for the lower problem of the bionical degree of glomerular model that exists at present, based on the glomerular chip that this application relates to, encapsulate the hollow fiber bundle in the bau chamber of chip with the mode that ". varies from" shape, vaccinate podocyte and endothelial cell respectively in the both sides of fibre pipe, thereby simulate glomerular capillary structure in vitro, realized the glomerular capillary blood vessel cluster and the bau chamber of separation, the physiological similarity of glomerular model has been improved. The filtrate outlet 6 on the chip 1 simulates the blood filtration behavior in the human body.

The embodiments in the present description are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments are referred 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 stacked from top to bottom in a first stacking manner;

a filtrate outlet channel (6), an arteriolar outlet channel (7), an arteriolar inlet channel (5) and a first liquid containing channel (8) communicated with the arteriolar inlet channel (5) are arranged on the first chip (1), the first liquid containing channel (8) is downward opened, and the filtrate outlet channel (6), the arteriolar outlet channel (7) and the arteriolar inlet channel (5) are not communicated with each other;

the second chip (2) is provided with a liquid channel, Bausch cavities (15), (16), (17), (18), (19), (20), a first through hole (21) of filtrate, a first outlet through hole (22), a first liquid containing groove (12), and a liquid conveying through hole (23) communicated with the Bausch cavities (15), (16), (17), (18), (19), (20), wherein the liquid containing groove (12) is communicated with the first liquid containing channel (8), the liquid channel comprises a arteriolar afferent microchannel (13) with an inlet connected with an outlet of the liquid containing groove (12) and a Bausch cavity inlet channel (14) communicated with the arteriolar afferent microchannel (13) and the Bausch cavities (15), (16), (17), (18), (19), (20);

the third chip (3) is provided with a second liquid containing groove (37), arteriolar efferent micro-channels (24), (25), (26), (27), (28), (29), an arteriolar outlet channel (30), Boehringer's through holes (31), (32), (33), (34), (35), (36) and a filtrate second through hole (38) communicated with the filtrate first through hole (21), the arteriolar efferent micro-channels (24), (25), (26), (27), (28), (29) and the arteriolar outlet channel (30) are communicated with the second liquid containing groove (37), the Boehringer's through holes (31), (32), (33), (34), (35) and (36) are communicated with the liquid conveying through hole (23), and the arteriolar efferent micro-channels (24), (25), (26), (27) and (28), (29) The Babys through holes (31), (32), (33), (34), (35), (36) and the second filtrate through hole (38) are not communicated with each other;

the fourth chip (4) is provided with a proximal tubule microchannel (39), (40), (41), (42), (43), (44), a third liquid containing groove (46) and a filtrate outlet channel (45) communicated with the third liquid containing groove (46), the third liquid containing groove (46) and the Bob's through holes (31), (32), (33), (34), (35), (36) are communicated with the proximal tubule microchannel (39), (40), (41), (42), (43), (44), and the 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 bauer cavities (15), (16), (17), (18), (19), (20) and the liquid channel are all plural, the bauer cavities (15), (16), (17), (18), (19), (20) and the liquid channel correspond to one another, and the liquid delivery through-hole (23) and the bauer cavities (15), (16), (17), (18), (19), (20) correspond to one another.

3. The glomerular chip of claim 2 wherein the Bowden through holes (31), (32), (33), (34), (35), (36) and the arteriolar efferent microchannels (24), (25), (26), (27), (28), (29) are all plural, and the Bowden through holes (31), (32), (33), (34), (35), (36) correspond one-to-one to the Bowden cavities (15), (16), (17), (18), (19), (20), and the arteriolar efferent microchannels (24), (25), (26), (27), (28), (29) correspond one-to-one to the Bowden cavity inlet channels (14).

4. The glomerular chip of claim 3 wherein the proximal tubule microchannels (39), (40), (41), (42), (43), (44) are plural, and the proximal tubule microchannels (39), (40), (41), (42), (43), (44) correspond to the Boehringer through holes (31), (32), (33), (34), (35), (36) one by one.

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 the cavities (15), (16), (17), (18), (19), (20) are identical in structure, and the cavities (15), (16), (17), (18), (19), (20) are respectively formed by the previous cavities (15), (16), (17), (18), (19), (20) at a predetermined angle with respect to the first liquid containing groove (12).

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

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

and/or the proximal tubule microchannels (39), (40), (41), (42), (43), (44) have a width of 0.5mm to 1.5mm and a depth of 0.3mm to 1.0 mm;

and/or the width of the filtrate outlet channel (45) is 0.5 mm-1.5 mm, and the depth is 0.3 mm-1.0 mm.

8. The glomerular chip of claim 1, wherein the inlet end of the arteriolar efferent microchannels (24), (25), (26), (27), (28), (29) is provided with a groove having a diameter greater than the width of the arteriolar efferent microchannels (24), (25), (26), (27), (28), (29).

9. The glomerular chip of claim 1 wherein the outlet end of the proximal tubule microchannel (39), (40), (41), (42), (43), (44) is provided with a groove having a diameter greater than the width of the proximal tubule microchannel (39), (40), (41), (42), (43), (44).

10. The glomerular chip of any of claims 1-9, wherein, in top-down projection:

the liquid 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;

and/or the outlet end through hole of the Babylonia cavity inlet channel (14) is respectively superposed with the circular grooves at the inlet ends of the arteriolar efferent microchannels (24), (25), (26), (27), (28) and (29);

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