Islet Isolation Device, Islet Isolation and Purification System, and Islet Isolation Method

Abstract

The present invention discloses an islet isolation device, an islet isolation and purification system, and an islet isolation method. The islet isolation device includes a digestive fluid circulation circuit and an islet isolation branch; the digestive fluid circulation circuit includes a pump and a digestion tank connected to each other through a circuit conduit; the islet isolation branch includes a first cell sieve and a separation tank connected to each other through a branch conduit. One end of the branch conduit is connected to the digestion tank through the first cell sieve, and the other end of the branch conduit is connected to the circuit conduit. A second cell sieve is also provided in the separation tank. The islet isolation device adds an islet isolation branch to the digestive fluid circulation circuit, which can smoothly retain islet cells in the separation tank. The separation and screening of the two-layer cell sieve can ensure the collection of most islets and reduce the total amount of harvested tissue. The islet isolation and purification system avoids the low yield problem of the density gradient centrifugation method, thereby effectively solving some defects of the existing method for separating islet cells by using the density gradient centrifugation technology.

Description

Technical Field

[0001] The present invention relates to the technical field of medical devices, and particularly to an islet isolation device, an islet isolation and purification system, and an islet isolation method. Background Art

[0002] Islets of Langerhans (pancreatic islets) are clusters of many cells, mainly sized at 100 - 300 μm, containing thousands of α cells, β cells, and δ cells, which are scattered in different parts of the pancreas. As the endocrine gland of the pancreas, the islets are responsible for secreting insulin, glucagon, etc. that can promote anabolism and regulate blood sugar stability. By regulating the secretion of insulin, glucagon, etc., the blood sugar in the body is maintained at a relatively normal fluctuating level. If the secretion of insulin is insufficient, diabetes will occur. Currently, the most ideal method for treating diabetes (especially type I diabetes) is cell therapy, that is, isolating islet cells from the pancreas and conducting corresponding tests and treatments. Developed countries around the world and developing countries including China have carried out related operations on islet cell isolation and transplantation, with remarkable curative effects, relieving diabetic patients of the pain of injecting insulin for many years.

[0003] The islet cells used in cell therapy are derived from donated human or porcine pancreases, and high-quality living cells are obtained by separating the pancreas of the donor. There are more than 6,000 donated pancreases in the country every year, and the number is increasing at a rate of 20% annually. However, in fact, less than 200 pancreases are successfully utilized each year, and the vast majority are abandoned, with a low utilization rate. The main reason is that it is relatively difficult to control the conditions of pancreatic digestion and separation.

[0004] Taking density gradient centrifugation, the main method currently used in the islet isolation and purification process, as an example, the main principle of this method is that the density of islet cell clusters is lower than that of exocrine tissue or ductal tissue. If these types of cells have been effectively separated during pancreatic digestion, then through the method of density gradient, sedimentation equilibrium can be achieved in the centrifugal force field, and a certain density gradient will appear from the liquid surface to the bottom in the sedimentation tank, thereby purifying the islet cell clusters.

[0005] The principle of islet cell separation by density gradient centrifugation is clear, and it can achieve specific separation of islet cells. However, in the actual use process, it is found that some exocrine gland tissues in the pancreas will have a density close to that of islet tissues due to reasons such as edema, which makes it difficult for the conventional COBE2991 continuous density gradient separation method to obtain a large number of high-purity islet cells. In addition, research has found that the components of the density gradient centrifugation liquid can cause the free islets to adhere to other pancreatic tissue cells during the density gradient centrifugation process, resulting in the failure to enrich some islet cells and the loss, causing a problem of low yield. At the same time, during the pancreatic digestion process, the islet cells released first will be broken into islet fragments by the steel balls for digestion and grinding in the digestion tank, further reducing the purity and efficiency of islet separation. Summary of the Invention

[0006] The present invention aims to solve at least one of the technical problems existing in the prior art. For this reason, the present invention provides an islet separation device, an islet separation and purification system, and an islet separation method that can improve the separation purity and separation efficiency.

[0007] In a first aspect, an embodiment of the present invention provides an islet separation device, which includes: a digestive juice circulation circuit and an islet separation branch; the digestive juice circulation circuit includes a pump and a digestion tank connected to each other through a circuit conduit; the islet separation branch includes a first cell sieve and a separation tank connected to each other through a branch conduit, one end of the branch conduit is connected to the digestion tank through the first cell sieve, the other end of the branch conduit is connected to the circuit conduit, and a second cell sieve is also provided in the separation tank; the pore size of the first cell sieve is 300 μm to 550 μm, and the pore size of the second cell sieve is 10 μm to 30 μm.

[0008] The islet separation device of the embodiment of the present invention has at least the following beneficial effects:

[0009] The islet isolation device provided by the present invention adds a branch for islet isolation to the digestive fluid circulation circuit. The pore size of the first cell sieve is from 300 μm to 550 μm, which can ensure that the digested islet cells can pass through; while the pore size of the second cell sieve is from 10 μm to 30 μm, which can smoothly retain the islet cells in the separation tank and prevent them from returning to the digestive fluid circulation circuit and being continuously digested, thus avoiding the occurrence of over-digestion or being broken into islet fragments by the grinding mechanism in the digestion tank. Islets, as independent endocrine gland structures in the pancreas, are different in size from the other tissue cells of the pancreas. The separation and screening by the two-layer cell sieve can ensure the collection of most islets, reduce the total tissue amount harvested, improve the purity of islets in the tissue, and avoid the low yield problem that the components of the density gradient centrifugation liquid can cause the adhesion of the already free islets to other pancreatic tissue cells during the density gradient centrifugation process, resulting in the loss of some islet cells not being enriched. Thus, some defects in the existing technology of separating islet cells by density gradient centrifugation are effectively solved.

[0010] According to another embodiment of the islet isolation device of the present invention, the separation tank is detachably connected to the branch conduit. The separation tank and the branch conduit are connected in a detachable manner, which can quickly replace the separation tank during the working process and avoid the accumulation of islet cells.

[0011] According to another embodiment of the islet isolation device of the present invention, there are at least two islet isolation branches. The setting of multiple islet isolation branches can ensure the efficiency of the separation work.

[0012] According to another embodiment of the islet isolation device of the present invention, an islet cell collection port is further provided on the separation tank. The provision of the islet cell collection port on the separation tank can timely collect the digested islet cells during the working process without replacing the separation tank.

[0013] According to another embodiment of the islet isolation device of the present invention, a heater is further connected to the circuit conduit, and the heater is used to heat the digestive fluid.

[0014] According to another embodiment of the islet isolation device of the present invention, the heater is a water bath, for example, it can be a water bath that can heat the water in the water storage cavity to 32 °C to 38 °C.

[0015] According to another embodiment of the islet isolation device of the present invention, a sampling tube is further connected to the circuit conduit.

[0016] According to another embodiment of the islet isolation device of the present invention, the sieve holes of the first cell sieve are 32, 35 or 40 mesh.

[0017] According to another embodiment of the islet isolation device of the present invention, the sieve holes of the second cell sieve are 425, 500, 625, 800 or 1250 mesh.

[0018] In a second aspect, an embodiment of the present invention provides an islet isolation and purification system, which includes the above-mentioned islet isolation device and a sorting workbench. The sorting workbench is used to further purify the enriched islet cell clusters in the separation tank.

[0019] For the islet isolation and purification system according to some other embodiments of the present invention, the sorting workbench is a magnetic bead sorting workbench.

[0020] During the process of using a COBE2991 cell separator to perform density gradient centrifugation for islet cell separation in the prior art, it was found that since the pancreas of a donor requires at least two COBE treatments, and each COBE takes about 40 minutes, this greatly increases the physical and mental consumption of the staff, and also makes the islets stay under non-culture conditions for a longer time, which is not conducive to maintaining the healthy state of the islets. If a similar workbench such as COBE2991 is selected in the islet isolation and purification system provided by the present invention, all the harvested tissues can be separated and purified by one COBE, significantly reducing the workload and being beneficial to maintaining the healthy state of islet cells. If the immunomagnetic bead sorting workbench of the present invention is used on the sorting workbench, it is possible to further avoid the problem of low yield caused by the adhesion of free islets to other pancreatic tissue cells during density gradient centrifugation due to the components of the density gradient centrifugation liquid, thus effectively solving some defects of the existing technology for separating islet cells using density gradient centrifugation.

[0021] For the islet isolation and purification system according to some other embodiments of the present invention, a magnetic bead sorting device is provided on the magnetic bead sorting workbench.

[0022] For the islet isolation and purification system according to some other embodiments of the present invention, the magnetic bead sorting workbench is a column-free magnetic isolation workbench.

[0023] For the islet isolation and purification system according to some other embodiments of the present invention, the magnetic bead sorting workbench includes a magnetic separator with a magnetic field. Within the magnetic field range of the magnetic separator, the islet cells bound to the antibody-labeled magnetic beads are adsorbed and retained, while other cells without specific surface antigens cannot bind to the antibody-labeled magnetic beads and have no magnetism, and are unable to be retained under the action of external force (or their own gravity), thus being separated from the islet cells bound to the antibody magnetic beads.

[0024] In a third aspect, an embodiment of the present invention provides an islet isolation method, which includes the following steps:

[0025] (1) Cutting the pancreas perfused with the digestive fluid into small pieces and mixing them with the digestive fluid for digestion;

[0026] (2) When free islet cells are detected in the digestive fluid, the digestive fluid is passed through a first cell sieve and a second cell sieve respectively. A cavity is formed between the first cell sieve and the second cell sieve, and the cell clusters in the cavity are collected.

[0027] (3) Sort the collected cell clusters.

[0028] Among them, the pore size of the first cell sieve is 300 μm to 550 μm, and the pore size of the second cell sieve is 10 μm to 30 μm.

[0029] According to another embodiment of the present invention, the sorting method is magnetic sorting. Magnetic sorting refers to a method in which, after applying an external magnetic field, other cells that are not bound to antibody magnetic beads are separated from the islet cells bound to antibody magnetic beads under the action of an external force (or their own gravity).

[0030] According to another embodiment of the present invention, the magnetic sorting is column-free magnetic isolation.

[0031] According to another embodiment of the present invention, the sorting method is density centrifugation. Specifically, a similar workbench such as a COBE2991 cell separator can be used for sorting.

[0032] Fourthly, an embodiment of the present invention provides a method for using the above-mentioned islet isolation and purification system, including the following steps:

[0033] (1) Cut the pancreas perfused with digestive fluid into small pieces and add them to the digestion tank together with the digestive fluid for mixed digestion.

[0034] (2) When free islet cells are detected in the digestive fluid, open the islet branch circuit, so that the digestive fluid flows in from the first cell sieve, passes through the separation tank and then flows out from the second cell sieve, thereby enriching cell clusters in the separation tank.

[0035] (3) Sort the enriched cell clusters on the sorting workbench.

[0036] According to another embodiment of the present invention, the sorting method for the above-mentioned islet isolation and purification system is magnetic sorting. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 is a schematic structural diagram of an islet isolation device in an embodiment of the present invention;

[0038] Figure 2 and Figure 3 is a partial schematic diagram of a column-free magnetic sorting device of a magnetic bead sorting workbench in an islet isolation and purification system in an embodiment of the present invention in a working state;

[0039] Figure 4 It is a schematic structural diagram of an islet isolation and purification system in another embodiment of the present invention. Specific Embodiments

[0040] The following will clearly and completely describe the concept of the present invention and the technical effects produced in combination with the embodiments, so as to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative efforts shall fall within the scope of protection of the present invention.

[0041] In the description of the embodiments of the present invention, if it involves orientation description, such as "upper", "lower", "front", "rear", "left", "right", etc., the orientation or position relationship indicated is based on the orientation or position relationship shown in the drawings, which is only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, so it cannot be understood as a limitation to the present invention.

[0042] In the description of the embodiments of the present invention, if a certain feature is referred to as "set", "fixed", "connected", "installed" on another feature, it can be directly set, fixed, connected, installed on another feature, or indirectly set, fixed, connected, installed on another feature. In the description of the embodiments of the present invention, if it involves "several", its meaning is more than one. If it involves "multiple", its meaning is more than two. If it involves "greater than", "less than", "exceeding", it should be understood as not including the number itself. If it involves "above", "below", "within", it should be understood as including the number itself. If it involves "first", "second", it should be understood as used to distinguish technical features, rather than indicating or implying relative importance or implicitly indicating the quantity of the indicated technical features or implicitly indicating the sequence relationship of the indicated technical features.

[0043] Embodiment 1

[0044] Reference Figure 1 , which shows a schematic structural diagram of an islet isolation device in an embodiment of the present invention. The islet isolation device includes a digestive fluid circulation circuit and an islet isolation branch. The digestive circulation circuit mainly includes a circuit conduit 140 and a pump 130, a digestion tank 110, a sampling tube 150 and a heater 160 that are sequentially connected to each other through the circuit conduit 140; the islet isolation branch mainly includes a branch conduit 123 and a first cell sieve 121 and a separation tank 122 that are connected to each other through the branch conduit 123. The circuit conduit 140 and the branch conduit 123 are at Figure 1It is characterized by different pipe diameters, which does not mean that the actual loop conduit 140 and the branch conduit 123 must have such a diameter relationship. In this embodiment, the heater 160 is a water bath. The loop conduit 140 enters its water storage cavity from one side of the water bath and leaves its water storage cavity from the other side of the water bath (not shown in the figure). A digestive fluid injection port 170 is provided on one side of the loop conduit 140. The digestive fluid injection port 170 can be externally connected to a container containing digestive fluid (not shown in the figure) to provide sufficient digestive fluid for the digestion of the pancreas. A digestive fluid regulating valve 171 is also provided on the digestive fluid injection port 170. By opening or closing the digestive fluid regulating valve 171, new digestive fluid is injected / stopped from being injected into the digestive fluid circulation loop and the pancreatic separation loop. The sampling tube 150 is sleeved in the loop conduit 140. The sampling tube 150 includes a housing 151 sleeved on the loop conduit 140 and a sampling tube 152 communicating with the housing 151 and perpendicular to the housing 151. A sampling tube switch 153 is provided on the sampling tube 152. The sampling tube 150 is used to detect the digestive fluid in the loop conduit 140 to confirm whether there are individual islet cells in the digestive fluid. The pump 130 can control the flow of the digestive fluid in the digestive fluid circulation loop and the islet separation branch. In this embodiment, the pump 130 is a peristaltic pump. The islet separation branch includes a first cell sieve 121 and a separation tank 122 connected to each other through a branch conduit 123. A second cell sieve 124 is provided in the separation tank 122. The first cell sieve 121 is provided on the upper surface of the digestion tank 110. One end of the branch conduit 123 is connected to the digestion tank 110 through the first cell sieve 121, and the other end of the branch conduit 123 is connected to the loop conduit 140. The separation tank 122 is connected to the branch conduit 123 through a first detachable joint 127 and a second detachable joint 128 provided on both sides. A first islet separation branch switch 125 is provided on the part of the branch conduit 123 between the first cell sieve 121 and the first detachable joint 127, and a second islet separation branch switch 126 is provided on the part of the branch conduit 123 close to the loop conduit 140. In this embodiment, there are two completely identical islet separation branches, that is, two completely identical first cell sieves 121 are provided on the upper surface of the digestion tank 110, and are respectively connected to the separation tanks 122 provided with second cell sieves 124 through the same branch conduits 123, and the opening or closing of a single islet separation branch is realized by the opening and closing of the first islet separation branch switch 125 and the second islet separation branch switch 126 provided on their respective branch conduits 123.

[0045] Reference Figure 2 and Figure 3, is a partial schematic diagram of the columnless magnetic separation device on the magnetic bead sorting workbench of the islet isolation and purification system according to an embodiment of the present invention in the working state. The columnless magnetic separation device includes a magnetic separator 210, which has a magnetic accommodation cavity for fixing a centrifuge tube 220.

[0046] The specific usage method of this islet separation device (i.e., the method for separating islet cells using this separation device) is as follows:

[0047] (1) After perfusing the digestive fluid into the pancreas, cut it into small pieces and put them into the digestion tank 110.

[0048] (2) Connect all the components of the islet separation device, turn on the heater 160 and the pump 130, open the digestive fluid regulating valve 171 on the digestive fluid injection port 170, and perfuse the digestive fluid into the entire separation device to start digestion.

[0049] (3) Regularly open the sampling tube switch 153 on the sampling tube 150 to sample the flowing digestive fluid and detect whether there are already separate free islet cells in it. If free islet cells have been detected, proceed to the next step; otherwise, continue digestion.

[0050] (4) Open a group of the first islet separation branch switches 125 and the second islet separation branch switches 126 to open one group of islet separation branches, so that the digestive fluid can pass through the islet separation branches; at the same time, open the digestive fluid regulating valve 171 to supplement the digestive fluid into the entire separation device. When the entire separation device is refilled with the digestive fluid, close the digestive fluid regulating valve 171.

[0051] (5) When there are a large number of cell clusters on the upper layer of the second cell sieve 124 in the separation tank 122, close the opened group of the first islet separation branch switches 125 and the second islet separation branch switches 126. At the same time, open another group of islet separation branches, so that the separation tank 122 in the other group of islet separation branches replaces the original working separation tank 122 to work. And open the digestive fluid regulating valve 171 to supplement the digestive fluid into the entire separation device.

[0052] (6) Drain the digestive fluid in the stopped working separation tank 122 and remove it, and wash the islet cell clusters remaining on the second cell sieve 124 in the separation tank 122 with the washing fluid for further separation work.

[0053] In this embodiment, the further separation work is to transfer the collected islet cell clusters to the magnetic bead sorting workbench for further separation work, which can refer to Figure 2 and Figure 3 , and the specific steps are as follows:

[0054] (1) Collect islet cell clusters with magnetic bead sorting buffer and transfer them to a centrifuge tube 220 for collection.

[0055] (2) Add antibody-labeled magnetic beads to the centrifuge tube 220, mix well with the collected islet cell clusters, and let stand for 10 minutes to allow the antibody-labeled magnetic beads to specifically bind to the islet cell clusters 310 sufficiently.

[0056] (3) Fix the centrifuge tube 220 in the accommodating cavity of the magnetic separator 210 and place it for 5 minutes.

[0057] (4) Flip the magnetic separator 210 so that the opening of the centrifuge tube 220 faces downward. Under the action of gravity, other types of cells or impurities 320 that fail to specifically bind to the antibody-labeled magnetic beads flow out of the centrifuge tube 220 under the action of gravity, while the islet cell clusters 310 that specifically bind to the antibody-labeled magnetic beads remain in the centrifuge tube 220 under the action of the magnetic field.

[0058] (5) Resuspend the islet cell clusters 310 in the centrifuge tube 220 to form a solution, and pass the solution through the separation tank 122. The complex of the islet cell clusters 310 and the antibody-labeled magnetic beads in the solution remains on the second cell sieve 124, while the magnetic beads that fail to bind to the islet cell clusters 310 flow out through the sieve holes of the second cell sieve 124.

[0059] Repeat the above steps until no islet cells can be detected in the digestive juice taken out by the sampling tube 150.

[0060] In this embodiment, the first cell sieve 121 is 40 mesh (sieve hole diameter 450 μm), and the second cell sieve 124 is 800 mesh (sieve hole diameter 15 μm).

[0061] In this embodiment, one of the separation tanks in the two sets of islet separation branches can be used and the other is reserved, or the two sets of separation tanks can be used alternately.

[0062] In this embodiment, the lower layer of the separation tank can recover the digestive juice, and the upper layer can enrich the target cell clusters. In the magnetic bead sorting workbench, the upper layer of the separation tank can enrich the cell clusters that bind to the antibody-labeled magnetic beads, while the magnetic beads that fail to bind to the islet cell clusters enter the lower layer of the separation tank.

[0063] In this embodiment, the digestive juice is removed through a separation tank, the cell mass is washed, and the solution is replaced, so that the islet cell mass is immersed in the magnetic bead sorting buffer solution, and the islet cells are statically combined with the antibody-labeled magnetic beads. In the existing islet cell magnetic bead sorting device, the antibody-labeled magnetic beads are directly added to the digestive juice in the digestion tank, and the antibody-labeled magnetic beads are combined with the islet cells in the digestion tank. However, the binding of the antibody-labeled magnetic beads to the specific markers on the cell surface needs to be carried out in the sorting buffer solution, and the environment of the digestive juice is obviously not suitable. In addition, the binding of the antibody-labeled magnetic beads to the cell surface markers requires a relatively static environment, and the violently shaking digestion tank obviously cannot make the antibody-labeled magnetic beads bind to the islet cells efficiently, which will lead to the excessive use of the antibody-labeled magnetic beads. The islet isolation and purification system provided by the present invention greatly reduces the dosage of the antibody-conjugated immunomagnetic beads and reduces the cost.

[0064] Example 2

[0065] An islet isolation device, which is different from that in Example 1 in that the setting of the separation tank is different. Refer to Figure 4 , which is a schematic diagram of the separation tank 122 of another embodiment of the present invention. The main body of the separation tank 122 is cylindrical, and a second cell sieve 124 is clamped inside. An islet cell collection port 129 is also provided on the side wall above the second cell sieve 124 in the separation tank 122 for transferring the enriched islet cell mass inside from this opening during use.

[0066] Example 3

[0067] An islet isolation and purification system, which is different from that in Example 1 in that it only includes a group of islet isolation branches.

[0068] Example 4

[0069] An islet isolation device, which is different from that in Example 1 in that a temperature sensor is provided on the digestion tank, and the temperature sensor can be connected to a display and / or processing platform through a wire, Bluetooth, wifi or other connection methods well known in the art to transmit the detected temperature data in the digestion tank to the display and / or processing platform for further processing or display. At the same time, the digestion time can be judged in combination with relevant data.

[0070] Example 5

[0071] An islet isolation device, which is different from that in Example 1 in that a third cell sieve is also provided at a position in the digestion tank close to the loop outlet (i.e., the outlet along the flow direction of the digestive juice), and the pore diameter of the third cell sieve is 1000-2000 μm. For example, it can be 9, 10, 12, 14, 16 meshes. By setting the third cell sieve, it can prevent tissues from entering the loop pipeline and causing blockage during digestion.

[0072] Example 6

[0073] An islet separation device is also provided with a circuit switch at a position where the circuit conduit is close to the digestion tank. When the digestion is stopped, the circuit switch is closed, and the digestive fluid can only flow into the islet separation branch, enabling it to quickly circulate in the branch and accelerating the separation efficiency of the separation tank.

[0074] The embodiments of the present invention have been described in detail above in conjunction with the accompanying drawings. However, the present invention is not limited to the above embodiments. Within the scope of knowledge possessed by those of ordinary skill in the art, various changes can be made without departing from the gist of the present invention. In addition, the embodiments of the present invention and the features in the embodiments can be combined with each other without conflict.

Claims

1. An islet isolation device, characterized in that, Comprising: A digestive fluid circulation circuit, including a pump and a digestion tank interconnected by a circuit conduit; An islet isolation branch, including a first cell sieve and a separation tank interconnected by a branch conduit, one end of the branch conduit being connected to the digestion tank through the first cell sieve, the other end of the branch conduit being connected to the circuit conduit, and a second cell sieve being further provided in the separation tank; the pore size of the first cell sieve is 300 μm to 550 μm, the pore size of the second cell sieve is 10 μm to 30 μm, and the separation tank is detachably connected to the branch conduit; Wherein, there are at least two of the islet isolation branches; the circuit conduit is further connected with a sampling tube.

2. The islet isolation device according to claim 1, characterized in that, An islet cell collection port is further provided on the separation tank.

3. The islet isolation device according to any one of claims 1 to 2, characterized in that, The circuit conduit is further connected with a heater for heating the digestive fluid.

4. An islet isolation and purification system, characterized in that, Comprising the islet isolation device according to any one of claims 1 to 3 and a sorting workbench for separating the cell mass enriched in the separation tank.

5. The islet isolation and purification system according to claim 4, wherein The sorting workbench is a column-free magnetic sorting workbench.

6. A method for islet isolation using the islet isolation device according to any one of claims 1 to 3 or the islet isolation and purification system according to any one of claims 4 to 5, characterized in that, Comprising the following steps: (1) Cutting the pancreas perfused with digestive fluid into small pieces and mixing them with the digestive fluid for digestion; (2) When free islet cells are detected in the digestive fluid, opening a group of islet isolation branches to enable the digestive fluid to pass through the first cell sieve and the second cell sieve respectively, a cavity being formed between the first cell sieve and the second cell sieve; when there are a large number of cell masses on the upper layer of the second cell sieve in the separation tank, closing a group of islet isolation branches, and at the same time opening another group of islet isolation branches, so that the separation tank in the other group of islet isolation branches replaces the separation tank in the original working state to work; and supplementing digestive fluid to the whole separation device; emptying the digestive fluid in the separation tank that has stopped working and removing it, and washing the islet cell mass remaining on the second cell sieve in the separation tank with a washing solution; (3) Sorting the collected islet cell mass; Wherein, the pore size of the first cell sieve is 300 μm to 550 μm, and the pore size of the second cell sieve is 10 μm to 30 μm.

7. The islet isolation method according to claim 6, characterized in that, The sorting is magnetic sorting.

8. A method for using the islet isolation and purification system according to any one of claims 4 to 5, characterized in that, Comprising the following steps: (1) Cutting the pancreas perfused with digestive fluid into small pieces and adding them together with the digestive fluid into the digestion tank for mixing and digestion; (2) When free islet cells are detected in the digestive fluid, opening a group of islet branch circuits to enable the digestive fluid to flow in from the first cell sieve, pass through the separation tank and flow out from the second cell sieve, a cavity being formed between the first cell sieve and the second cell sieve; when there are a large number of cell masses on the upper layer of the second cell sieve in the separation tank, closing a group of islet isolation branches, and at the same time opening another group of islet isolation branches, so that the separation tank in the other group of islet isolation branches replaces the separation tank in the original working state to work; and supplementing digestive fluid to the whole separation device; emptying the digestive fluid in the separation tank that has stopped working and removing it, and washing the islet cell mass remaining on the second cell sieve in the separation tank with a washing solution; (3) Sorting the islet cell mass enriched on the sorting workbench.

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