CN113621515B - Glomerular chip - Google Patents

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 glomerulus chip.

Background technique

The kidney is an important organ of the human body with an extremely complex structure and microenvironment. The kidney is composed of millions of nephrons, and the glomerulus is an important part of the nephron. The glomerulus is responsible for blood filtration, and its filtration membrane is a three-layer structure with a basement membrane in the middle. Glomerular filtration is the initial process of urine formation. When blood flows through the glomerulus, most substances are filtered by glomerular capillaries and enter Bowman's cavity to form primary urine. Research has shown that glomerular dysfunction is the cause of most kidney diseases such as chronic kidney disease, diabetic nephropathy, and nephrotic syndrome. Therefore, researchers are committed to establishing in vitro glomerular models to reveal the mechanism of glomerular function.

Currently, there are two main types of glomerular models in vitro: kidney organoids and glomerular chips. The glomeruli constructed from kidney organoids can express podocyte-associated proteins and have glomerular filtration function. However, the existing kidney organoids have not realized the spherical glomerular structure with Bowman's cavity and lack of perfusable vascular network, so its application is limited. The glomerulus chip is an in vitro glomerular model based on emerging micro-engineering technologies (such as micro-fabrication, microfluidics, and 3D printing technologies). In recent years, glomerular chips based on materials such as polydimethylsiloxane (PDMS) and organic glass (PMMA) combined with extracellular matrix such as hydrogel and decellularized tissue have been widely reported. However, the basement membrane of the existing glomerular chips is flat rather than spherical and twisted, resulting in limited glomerular filtration function. In conclusion, current in vitro glomerular models are still relatively low in biomimeticity.

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

Contents of the invention

The purpose of the present invention is to provide a glomerulus chip, and the bionicity of the glomerulus chip is improved.

To achieve the above object, the present invention provides a glomerulus chip, comprising a first chip, a second chip, a third chip and a fourth chip stacked sequentially 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 first liquid accommodating channel The opening of the channel is downward, and the first filtrate outlet channel, the first arteriole outlet channel and the arteriole inlet channel are not connected to each other;

The second chip is provided with a liquid channel, a Bowman cavity, a first through hole for filtrate, a first outlet through hole, a first liquid storage tank, and a liquid delivery through hole communicating with the Bowman cavity. The first liquid storage tank is in communication with the first liquid storage channel, and the liquid channel includes an arteriolar afferent microchannel whose inlet is connected to the outlet of the first liquid storage tank and communicates with the arteriolar afferent microchannel. channel and the Bowman cavity inlet channel of the Bowman cavity, the Bowman cavity is used to accommodate the hollow fiber, and the two ends of the hollow fiber are respectively embedded in the Bowman cavity inlet channel, so that the culture medium can only pass through the hollow the inner lumen of the fiber;

The third chip is provided with a second liquid holding tank, an arteriole efferent microchannel, a second arteriole outlet channel, a Bowman through hole, and a second through hole of the filtrate connected to the first through hole of the filtrate, The arteriole efferent microchannel and the second arteriole outlet channel are both in communication with the second liquid holding tank, the Bowman through hole is in communication with the liquid delivery through hole, and the arteriole efferent The microchannel is in communication with the entrance channel of the Bowman's cavity, and the entrance of the microchannel efferent from the arteriole is connected with the through hole at the outlet end of the entrance channel of the Bowman's cavity, and the through hole of the Bowman's cavity is connected with the filtrate second The through holes are not connected to each other;

The fourth chip is provided with a third liquid holding tank of the proximal tubule microchannel and a second filtrate outlet passage communicated with the third liquid holding tank, the third liquid holding tank and the abalone The through holes are all communicated with the proximal tubule microchannel, and the second filtrate outlet channel is communicated with the second through hole of the filtrate.

Preferably, the liquid delivery through hole, the Bowman cavity and the liquid channel are multiple, the Bowman cavity corresponds to the liquid channel one by one, and the liquid delivery through hole and the Bowman cavity Cavity one to one correspondence.

Preferably, both the Bowman's through holes and the arteriolar efferent microchannels are multiple, and the Bowman's through holes correspond to the Bowman's lumens one by one, and the arteriolar efferent microchannels correspond to the Bowman's lumens one by one. There is a one-to-one correspondence between the inlet channels of Bowman's cavity.

Preferably, there are multiple proximal tubule microchannels, and the proximal tubule microchannels are in one-to-one correspondence with the Bowman through holes.

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

Preferably, there are multiple Bowman cavities with the same structure, wherein each of the Bowman cavities is obtained from the previous Bowman cavity with the first liquid storage tank as a preset angle.

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

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

And/or the microchannel of the proximal tubule has a width of 0.5 mm to 1.5 mm and a depth of 0.3 mm to 1.0 mm;

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

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 with a diameter larger than the width of the proximal tubule microchannel.

Preferably, from top to bottom projection:

The first liquid storage channel, the first liquid storage tank, the second liquid storage tank and the third liquid storage tank overlap;

And/or the through hole at the outlet end of the Bowman's cavity inlet channel coincides with the circular groove at the inlet end of the efferent microchannel of the arteriole respectively;

And/or the liquid conveying through holes coincide with the Bowman through holes respectively.

In the above technical solution, the glomerulus 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 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 Bowman cavity, a filtrate first through hole, and a first outlet through hole , a first liquid storage tank, a liquid delivery through hole communicating with Bowman's cavity, the liquid storage tank communicates with the first liquid storage channel, and the liquid channel includes an arteriolar afferent microchannel whose inlet is connected with the outlet of the liquid storage tank and the Bowman's cavity inlet channel connecting the arteriole afferent microchannel and Bowman's cavity; the third chip is provided with a second liquid holding tank, an arteriole efferent microchannel, an arteriole outlet channel, a Bowman through hole and a connecting hole. The second through hole of the filtrate passing through the first through hole of the filtrate, the arteriole efferent microchannel and the arteriole outlet channel are all connected with the second liquid holding tank, the Bowman through hole is connected with the liquid delivery through hole, and the arteriole efferent The microchannel communicates with the inlet channel of the Bowman cavity; the fourth chip is provided with a proximal tubule microchannel, a third liquid storage tank and a filtrate outlet channel connected with the third liquid storage tank, the third liquid storage tank and The Bowman through holes are all connected with the microchannel of the proximal tubule, and the outlet channel of the filtrate is connected with the second through hole of the filtrate.

From the above description, it can be seen that in the glomerular chip provided by the present application, by setting the Bowman cavity, a functional area is provided for the loading of glomerular capillaries into the Bowman cavity on the chip, thereby forming a bionic glomerular capillary Vascular-like structure, which in turn improves the biomimeticity of the glomerulus-on-a-chip.

Description of drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings that need to be used in the description of the embodiments or the prior art. Obviously, the accompanying drawings in the following description are only It is an embodiment of the present invention, and those skilled in the art can also obtain other drawings according to the provided drawings without creative work.

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

FIG. 2 is a schematic structural diagram of a first chip provided by an embodiment of the present invention;

FIG. 3 is a schematic structural diagram of a second chip provided by an embodiment of the present invention;

FIG. 4 is a schematic structural diagram of a third chip provided by an embodiment of the present invention;

FIG. 5 is a schematic structural diagram of a fourth chip provided by an embodiment of the present invention.

Among them, 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-the first Small artery outlet channel, 8-first liquid holding channel, 9-screw hole, 10-screw hole, 11-screw hole, 12-first liquid holding tank, 13-arteriole afferent microchannel, 14- Bowman's cavity inlet channel, 15-Bowman's cavity, 16-Bowman's cavity, 17-Bowman's cavity, 18-Bowman's cavity, 19-Bowman's cavity, 20-Bowman's cavity, 21-the first through hole of filtrate , 22-first outlet through hole, 23-liquid delivery through hole, 24-arteriole efferent microchannel, 25-arteriole efferent microchannel, 26-arteriole efferent microchannel, 27-arteriole efferent microchannel Channel, 28-arteriole efferent microchannel, 29-arteriole efferent microchannel, 30-second arteriole outlet channel, 31-Bowman's through hole, 32-Bowman's through hole, 33-Bowman's through hole, 34-Bowman through hole, 35-Bowman through hole, 36-Bowman through hole, 37-second liquid holding tank, 38-filter liquid second through hole, 39-proximal tubule microchannel, 40-near Terminal tubule microchannel, 41-proximal tubule microchannel, 42-proximal tubule microchannel, 43-proximal tubule microchannel, 44-proximal tubule microchannel, 45-second filtrate outlet channel, 46-third Liquid holding tank.

Detailed ways

The core of the present invention is to provide a glomerulus chip, and the bionicity of the glomerulus chip is improved.

In order to enable those skilled in the art to better understand the technical solutions of the present invention, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.

Please refer to Figure 1 to Figure 5.

In a specific embodiment, the glomerulus chip provided by the specific 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 circular, rounded rectangle or rectangle, the first chip 1, the second chip 2, the third chip 3 and the fourth chip 4 The areas of the upper surface and the lower surface are equal, and the specific area may be 110 mm in length and 90 mm in width. The thickness of the first chip 1 should not be too thin, generally greater than 8mm, specifically, it can be 10mm, which is convenient for drilling screw holes on the side of the chip. If the first chip 1 is too thin, the culture medium will leak during the operation of the chip. The thickness of the second chip 2 and the third chip 3 is 2 mm to 4 mm, which should not be too thick or too thin. If the middle two-layer chip is too thick, it is not conducive to the assembly of the entire glomerulus chip; if the middle two-layer chip is too thin, it will be easily bent and deformed during chip assembly. The thickness of the fourth chip 4 is generally greater than 3 mm, specifically 4 mm. If it is too thin, it will be easily bent and deformed, and the culture medium will easily leak during the 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 can be organic glass (PMMA) or polydimethylsiloxane (PDMS); in this embodiment There are no special restrictions on the type and source of the plexiglass and PDMS, and the plexiglass and PDMS well known to those skilled in the art can be used, which can be purchased from the market; as in the embodiments of the present invention, Suzhou Anheda can be used. Plexiglass from Plastic Products Co. or PDMS polymer from Corning, Midland, Michigan, USA.

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 can also be fastened by buckles. Specifically, the first chip 1 is provided with screw holes 9, 10, 11, and the screw holes 9, 10, 11 are arranged circumferentially around the first chip 1, specifically ten. The second chip 2, the third chip 3 and The fourth chips 4 are respectively provided with mounting holes corresponding to the screw holes 9 , 10 , 11 so as to be fixed as a whole.

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 accommodating channel 8 communicated with the arteriole inlet channel 5, the first liquid accommodating channel The opening of 8 is downward, specifically, the arteriole inlet 5 communicates with the first liquid containing channel 8 at the bottom of the first layer chip 1 . The three channels of the arteriole inlet channel 5, the first filtrate outlet channel 6 and the first arteriole outlet channel 7 are set independently, that is, they do not intersect with each other, and the two are not connected. Specifically, the arteriole inlet channel 5 is responsible for transporting the medium and perfusing the cell suspension, the first filtrate outlet channel 6 is responsible for collecting the medium that has not been filtered by the glomerulus, and the first arteriole outlet channel 7 is responsible for collecting the glomerulus Filtered filtrate.

The second chip is provided with liquid channels, Bowman chambers 15, 16, 17, 18, 19, 20, filtrate first through hole 21, first outlet through hole 22, first liquid holding tank 12, and Bowman Cavities 15, 16, 17, 18, 19, 20 communicate with the liquid delivery through hole 23, the first liquid storage tank 12 communicates with the first liquid storage channel 8, and the liquid channel includes an inlet and an outlet of the first liquid storage tank 12 The connected arteriolar afferent microchannel 13 and the Bowman's lumen inlet channel 14 connecting the arteriolar afferent microchannel 13 and the Bowman's lumens 15 , 16 , 17 , 18 , 19 , 20 . Specifically, there can be one or at least two Bowman's cavities 15, 16, 17, 18, 19, and 20. As shown in the figure, there are six Bowman's cavities, namely, Bowman's cavity 15, Bowman's cavity 16, 17 Bowman's cavity, 18 Bowman's cavity, 19 Bowman's cavity and 20 Bowman's cavity.

When the liquid delivery through hole 23, the Bowman cavity 15, 16, 17, 18, 19, 20 and the liquid channel are multiple, the Bowman cavity 15, 16, 17, 18, 19, 20 corresponds to the liquid channel one by one, The liquid delivery through hole 23 is in one-to-one correspondence with the Bowman cavity 15, 16, 17, 18, 19, and 20. Specifically, the liquid delivery through hole 23, the arteriole afferent microchannel 13 and the Bowman cavity inlet channel 14 are all connected to the Bowman cavity. The number of Cavities 15, 16, 17, 18, 19, and 20 is equal.

Specifically, the first liquid holding tank 12 communicates with the opening at the bottom end of the first liquid holding channel 8 to receive the culture medium flowing in from the arteriole inlet channel 5 in the first chip 1; the first liquid holding tank 12 is connected to the small artery The incoming microchannel 13 communicates with the Bowman cavity inlet channel 14, and the culture medium is delivered to the Bowman cavity 15, 16, 17, 18, 19, 20; the Bowman cavity 15, 16, 17, 18, 19 and 20 are The main functional area of the entire chip for the loading of glomerular capillary clusters.

There are multiple Bowman cavities 15, 16, 17, 18, 19, and 20 with the same structure, wherein the Bowman cavities 15, 16, 17, 18, 19, and 20 are formed by the previous Bowman cavity 15, 16, and 17 respectively. , 18, 19, 20 are obtained by taking the first liquid holding tank 12 as a preset angle. Specifically, the diameter of the Bowman cavity 15, 16, 17, 18, 19, and 20 is 10 mm, and the depth is 1 mm. The groove 12 is obtained by rotating the center of circle by 36°. The Bowman's cavity inlet channel 14 tangent to the Bowman's cavity 15, 16, 17, 18, 19, 20 has a width of 3 mm, and the fluid channel depth of the Bowman's cavity inlet channel 14 is 1 mm.

The third chip is provided with a second liquid holding tank 37, arteriole efferent microchannels 24, 25, 26, 27, 28, 29, a second arteriole outlet channel 30, Bowman through holes 31, 32, 33, 34,35,36 and the filtrate second through hole 38 of the first through hole 21 connected with the filtrate, the arteriole efferent microchannel 24,25,26,27,28,29 and the second arteriole outlet channel 30 are all connected with The second liquid holding tank 37 communicates, the Bowman through holes 31, 32, 33, 34, 35, 36 communicate with the liquid delivery through hole 23, and the arteriole efferent microchannels 24, 25, 26, 27, 28, 29 communicate with the The Bowman's cavity inlet channel 14 communicates. Preferably, Bowman's through holes 31, 32, 33, 34, 35, 36 and arteriolar efferent microchannels 24, 25, 26, 27, 28, 29 are multiple, and Bowman's through holes 31, 32, 33 . One to one correspondence.

Specifically, the arteriole efferent microchannels 24, 25, 26, 27, 28, 29 on the third chip 3 and the second arteriole outlet channel 30 are all communicated with the second liquid holding tank 37; 31 , 32 , 33 , 34 , 35 , 36 and the second through hole 38 of filtrate are set independently and do not communicate with each other, that is, they do not intersect each other. The inlets of the arteriole efferent microchannels 24 , 25 , 26 , 27 , 28 and 29 are connected to the through hole at the outlet end of the inlet channel 14 of Bowman's lumen. Specifically, the arteriole efferent microchannels 24, 25, 26, 27, 28 and 29 collect medium from the second chip 2 that has not been filtered by the glomeruli, converge on the second arteriole outlet channel 30 and pass through the second chip 2. The first outlet on the second chip 2 flows out.

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

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

The fourth chip is provided with proximal tubule microchannels 39, 40, 41, 42, 43, 44, a third liquid holding tank 46 and a second filtrate outlet channel 45 communicated with the third liquid holding tank 46. Three liquid holding tanks 46 and Bowman through holes 31, 32, 33, 34, 35, 36 are all communicated with the proximal tubule microchannel 39, 40, 41, 42, 43, 44, and the second filtrate outlet channel 45 is connected with The filtrate is communicated with the second through hole 38 .

The proximal tubule microchannels 39 , 40 , 41 , 42 , 43 , 44 and the second filtrate outlet channel 45 are all in communication with the third liquid holding tank 46 . Specifically, the length of the proximal tubule microchannels 39, 40, 41, 42, 43, and 44 is 41.5 mm, and the width of the proximal tubule microchannels 39, 40, 41, 42, 43, and 44 is 0.5 mm to 1.5 mm, which can be specifically 1mm, the depth is 0.3mm-1.0mm, specifically 0.5mm, the second filtrate outlet channel 45 is 30mm long, 0.5mm-1.5mm wide, specifically 1mm, and the depth is 0.3mm-1.0mm, specifically 0.5mm mm. The medium filtered by the glomerulus is collected on the second filtrate outlet channel 45 through the proximal tubule microchannels 39, 40, 41, 42, 43, 44 and finally passes through the second through hole of the filtrate on the third chip 3 38 and the first through hole 21 of the filtrate on the second chip 2 returns to the first chip 1 .

As shown in the figure, there are multiple proximal tubule microchannels 39, 40, 41, 42, 43, 44. 33, 34, 35, 36 correspond to each other.

The inlet ends of the arteriolar efferent microchannels 24 , 25 , 26 , 27 , 28 , and 29 are provided with grooves whose diameter is larger than the width of the arteriolar efferent microchannels 24 , 25 , 26 , 27 , 28 , and 29 .

The outlet ends of the proximal tubule microchannels 39 , 40 , 41 , 42 , 43 , 44 are provided with grooves whose diameters are larger than the width of the proximal tubule microchannels 39 , 40 , 41 , 42 , 43 , and 44 .

On the basis of the above schemes, project 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 overlap.

The through hole at the outlet end of the Bowman's lumen inlet channel 14 coincides with the circular grooves at the inlet ends of the arteriole efferent microchannels 24 , 25 , 26 , 27 , 28 , and 29 respectively.

The liquid delivery through hole 23 coincides with the Bowman through holes 31 , 32 , 33 , 34 , 35 , and 36 respectively.

The assembly method of the glomerulus chip design can be: arrange the first chip 1, the second chip 2, the third chip 3, and the fourth chip 4 from top to bottom in sequence. Make the first liquid holding channel 8 on the bottom of the first chip 1 coincide with the first liquid holding groove on the top of the second chip 2; The first outlet through hole 22 coincides; the hole of the first filtrate outlet channel 6 at the bottom of the first chip 1 coincides with the first through hole 21 of the filtrate on the second chip 2; the first chip 1 can be determined by the alignment of these structures and the relative position of the second chip 2. Then, make the terminal through-holes of the Bowman's lumen inlet channel 14 on the second chip 2 coincide with the front circular grooves of the arteriole efferent microchannels 24, 25, 26, 27, 28, 29 on the third chip 3 respectively; Make the filtrate first through hole 21 on the second chip 2 coincide with the filtrate second through hole 38 on the third chip 3; Make the first outlet through hole 22 on the second chip 2 and the third chip 3 The round holes at the ends of the second arteriole outlet channel 30 coincide; the relative positions of the second chip 2 and the third chip 3 can be determined through the alignment of these structures. Then, the second liquid holding groove 37 on the third chip 3 is aligned with the third liquid holding groove 46 on the fourth chip 4, and the Bowman through holes 31, 32, 33, 34 on the third chip 3 are aligned. , 35, 36 are respectively aligned with the inlet circular grooves of the proximal tubule microchannel 44, 43, 42, 41, 40, 39 on the fourth chip 4; the filtrate second through hole 38 on the third chip 3 is aligned with The circular grooves at the outlet end of the second filtrate outlet channel 45 on the fourth chip 4 are aligned; through the 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 glomerulus chip design can be completed . Specifically, the first chip 1 , the second chip 2 , the third chip 3 and the fourth chip 4 can be fastened by 10 screws at the edges. Specifically, between the first chip 1 and the second chip 2, between the second chip 2 and the third chip 3, and between the third chip 3 and the fourth chip 4, a layer of silica gel film can be sandwiched respectively to improve the chip of tightness.

The glomerulus chip provided by this application is designed to construct a glomerulus model in vitro, and the specific process is as follows:

Wrap the hollow fiber end (~3cm) taken from the plasma separator with an equal amount of silica gel, and then put it in a drying oven at 60°C for 20 minutes until the silica gel is completely dry; place the dried hollow fiber along the silica gel wrapped part Cut in the middle to expose the hollow channel, and bend it into a "∝"shape; place the bent hollow fiber in the Bowman cavity 15, 16, 17, 18, 19, 20 on the second chip 2, specifically , the main part of the hollow fiber bending is placed in the Bowman cavity 15, 16, 17, 18, 19 and 20, and the two ends are respectively embedded in the Bowman cavity inlet channel 14 on the second chip 2, and fixed with silica gel, so that the culture medium It can only pass through the inner lumen of the hollow fiber, thereby simulating the cluster structure of human glomerular capillaries; the hollow fiber is coated with human fibronectin (0.5mg/mL; directly prepared with sterile PBS) for 1 hour (37°C , 5% CO ₂ ) to support the adhesion and growth of podocytes; inoculate 30 μL of podocyte suspension (1×10 ⁷ cells/mL) into the hollows of Bowman chambers 15, 16, 17, 18, 19, and 20 fiber surface, and incubated in the incubator for two hours to promote cell adhesion on the surface of the hollow fiber tube. A silicone resin film (thickness: ~0.5 mm) was sandwiched between every two layers of chips, and the chip assembly process was completed as described. Using a 5 mL syringe, 1 mL of endothelial cell suspension (2×10 ⁵ cells/mL) was perfused into the hollow fiber through the arteriole inlet 5 on the first chip 1 , and allowed to stand for 2 hours to support the adhesion of endothelial cells on the inner surface of the hollow fiber. Connect a sterile peristaltic tube to a medium bottle containing 60 mL of cell culture medium and place in the incubator. The medium is driven by a peristaltic pump at a speed of 0.2 mL/h, passes through the arteriole inlet 5 on the first chip 1, and then enters the fiber tubes in the six Bowman lumens 15, 16, 17, 18, 19 and 20 , flow through the microchannels 24, 25, 26, 27, 28 and 29 from the arterioles on the third chip 3, and finally return to the culture medium bottle from the arteriole outlet 7 on the first chip 1, and another part of the culture medium is abalone Fibrous tubes (glomerular capillary structures) in the lumen 15, 16, 17, 18, 19 and 20 are filtered and then flow through the proximal tubule microchannels 39, 40, 41, 42, 43, 44 and return to the filtrate outlet 6 on the first chip 1, simulating the blood filtration behavior in the human body.

This application can realize the simulation of glomerular capillary structure in vitro, and can realize separated capillary clusters and Bowman's cavity at the same time, and provide solutions for the problems of low bionic degree of glomerular models currently existing, based on the aspects involved in this application The glomerular chip encapsulates the hollow fiber bundle in the Bowman cavity of the chip in a "∝" shape, and inoculates podocytes and endothelial cells on both sides of the fiber tube, thereby simulating the glomerular capillary structure in vitro and realizing Isolated glomerular capillary tufts and Bowman's lumen improve the physiological similarity of the glomerular model. The filtrate outlet 6 on the chip 1 simulates the blood filtration behavior in the human body.

Each embodiment in this specification is described in a progressive manner, each embodiment focuses on the difference from other embodiments, and the same and similar parts of each embodiment can be referred to each other.

The above description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the present invention will not 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 (10)

1. A glomerulus chip, characterized in that it comprises a first chip (1), a second chip (2), a third chip (3) and a fourth chip (4) stacked sequentially 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 communicates with the arteriole inlet channel (5). The first liquid accommodating channel (8), the opening of the first liquid accommodating channel (8) is downward, 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 Bowman chamber (15), (16), (17), (18), (19), (20), a filtrate first through hole (21) , the first outlet through hole (22), the first liquid holding tank (12), communicate with the Bowman cavity (15), (16), (17), (18), (19), (20) The liquid delivery through hole (23), the first liquid storage tank (12) communicates with the first liquid storage channel (8), and the liquid channel includes an inlet and the first liquid storage tank ( 12) The arteriolar afferent microchannel (13) connected to the outlet and the arteriolar afferent microchannel (13) connected with the Bowman lumen (15), (16), (17), (18), ( 19), (20) Bowman cavity inlet channels (14), said Bowman cavity (15), (16), (17), (18), (19), (20) are used to accommodate hollow fibers, And the two ends of the hollow fiber are respectively embedded in the inlet channel (14) of the Bowman cavity, so that the culture medium can only pass through the inner lumen of the hollow fiber; The third chip (3) is provided with a second liquid holding tank (37), arteriole efferent microchannels (24), (25), (26), (27), (28), (29) , the second arteriole outlet channel (30), Bowman's through hole (31), (32), (33), (34), (35), (36) and the first through hole (21) communicating with the filtrate ) of the filtrate second through hole (38), the arteriole efferent microchannel (24), (25), (26), (27), (28), (29) and the second arteriole The outlet passages (30) are all in communication with the second liquid holding tank (37), and the Bowman through holes (31), (32), (33), (34), (35), (36) are connected with The liquid delivery through hole (23) communicates, and the arteriole efferent microchannels (24), (25), (26), (27), (28), (29) are connected with the Bowman cavity inlet channel (14) is communicated, and the inlet of described arteriole efferent microchannel (24), (25), (26), (27), (28), (29) is connected with described Bowman's lumen inlet channel (14) The outlet end through hole, the Bowman through holes (31), (32), (33), (34), (35), (36) and the second through hole of the filtrate (38) do not communicate with each other ; The fourth chip (4) is provided with proximal tubule microchannels (39), (40), (41), (42), (43), (44), the third liquid holding tank (46) and The second filtrate outlet channel (45) communicating with the third liquid storage tank (46), the third liquid storage tank (46) and the Bowman through holes (31), (32), (33), (34), (35), (36) are all communicated with described proximal tubule microchannel (39), (40), (41), (42), (43), (44), so The second filtrate outlet channel (45) communicates with the second filtrate through hole (38). 2. The glomerulus chip according to claim 1, characterized in that, the liquid delivery through hole (23), the Bowman cavity (15), (16), (17), (18), ( 19), (20) and the liquid channel are multiple, and the Bowman cavity (15), (16), (17), (18), (19), (20) and the liquid channel one One-to-one correspondence, the liquid delivery through hole (23) corresponds to the Bowman cavity (15), (16), (17), (18), (19), (20) one-to-one. 3. The glomerulus chip according to claim 2, characterized in that, the Bowman through holes (31), (32), (33), (34), (35), (36) and the Arteriole efferent microchannels (24), (25), (26), (27), (28), (29) are all multiple, and the Bowman's through holes (31), (32), ( 33), (34), (35), (36) are in one-to-one correspondence with the Bowman cavity (15), (16), (17), (18), (19), (20), and the small The arterial efferent microchannels (24), (25), (26), (27), (28), (29) correspond one-to-one to the Bowman's lumen inlet channel (14). 4. The glomerulus chip according to claim 3, characterized in that, the proximal tubule microchannels (39), (40), (41), (42), (43), (44) are multiple One, the proximal tubule microchannel (39), (40), (41), (42), (43), (44) and the Bowman through hole (31), (32), (33) , (34), (35), (36) one-to-one correspondence. 5. The glomerulus chip according to claim 1, wherein the first chip, the second chip, the third chip and the fourth chip are fastened by threaded fasteners. 6. The glomerulus chip according to claim 1, characterized in that, there are multiple Bowman's cavities (15), (16), (17), (18), (19), (20), And the structure is the same, wherein, the Bowman cavity (15), (16), (17), (18), (19), (20) respectively by the previous said Bowman cavity (15), (16) , (17), (18), (19), (20) are obtained with the first liquid holding tank (12) as a preset angle. 7. glomerulus chip according to claim 1, is characterized in that, described arteriole efferent microchannel (24), (25), (26), (27), (28), (29) The width is 0.5mm~1.5mm, and the depth is 0.3mm~1.0mm; And/or the second arteriole outlet channel (30) has a width of 0.5 mm to 1.5 mm and a depth of 0.3 mm to 1.0 mm; And/or the proximal tubule microchannels (39), (40), (41), (42), (43), (44) have a width of 0.5 mm to 1.5 mm and a depth of 0.3 mm to 1.0 mm; And/or the second filtrate outlet channel (45) has a width of 0.5mm-1.5mm and a depth of 0.3mm-1.0mm. 8. The glomerulus chip according to claim 1, characterized in that, the arteriole efferent microchannel (24), (25), (26), (27), (28), (29) The inlet end is provided with a groove whose diameter is larger than the width of the arteriolar efferent microchannels (24), (25), (26), (27), (28), (29). 9. glomerulus chip according to claim 1, is characterized in that, the outlet end of proximal tubule microchannel (39), (40), (41), (42), (43), (44) is provided with There are grooves with a diameter greater than the width of the proximal tubule microchannels (39), (40), (41), (42), (43), (44). 10. The glomerulus chip according to any one of claims 1-9, characterized in that, projected from top to bottom: The first liquid storage channel (8), the first liquid storage tank (12), the second liquid storage tank (37) and the third liquid storage tank (46) overlap; And/or the through hole at the outlet end of the Bowman's lumen inlet channel (14) is connected with the efferent microchannel of the arteriole (24), (25), (26), (27), (28), (29) respectively The circular grooves at the inlet end coincide; And/or the liquid delivery through hole (23) coincides with the Bowman through hole (31), (32), (33), (34), (35), (36) respectively.

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