CN111607508B

CN111607508B - Automatic mesenchymal stem cell separation device

Info

Publication number: CN111607508B
Application number: CN202010347495.0A
Authority: CN
Inventors: 徐建勇; 吴慧玫
Original assignee: Shenzhen University
Current assignee: Shenzhen University
Priority date: 2020-04-28
Filing date: 2020-04-28
Publication date: 2023-03-10
Anticipated expiration: 2040-04-28
Also published as: CN111607508A

Abstract

The invention provides an automatic mesenchymal stem cell separation device which comprises a shell, a controller arranged outside the shell, a tissue separator arranged inside the shell, and a motor used for driving the tissue separator to operate, wherein a plurality of liquid storage tanks are formed between the tissue separator and the shell, a liquid inlet pipeline and a liquid discharge pipeline are arranged below the liquid storage tanks, the liquid storage tanks are connected with the liquid inlet pipeline and the liquid discharge pipeline through a communicating pipe with an electromagnetic valve, the liquid inlet pipeline and the liquid discharge pipeline are connected with the tissue separator through a liquid pump, and the motor, the electromagnetic valve and the liquid pump are all electrically connected with the controller. The mesenchymal stem cell separation device is compact in structure, and after an operation program is set by the controller, the tissue block to be extracted only needs to be placed in the tissue separator, so that the device can automatically cut and extract the tissue block in a sample adding manner, and can automatically clean and sterilize the device after extraction is finished, thereby saving manpower and improving the separation efficiency of mesenchymal stem cells.

Description

Automatic mesenchymal stem cell separation device

Technical Field

The invention relates to the technical field of regenerative medicine experimental equipment, in particular to an automatic mesenchymal stem cell separation device.

Background

Stem cell research and regenerative medicine research have become the leading edge and hot spot of today's biomedical research. Adult stem cells are undifferentiated stem cells existing in adult tissues, and are widely used in the fields of regenerative biology and regenerative medicine because they can differentiate into specific cell types, repair tissues, and maintain the integrity of tissues. Adult stem cells having a wide range of application values in the human body include hematopoietic stem cells and mesenchymal stem cells. Among them, mesenchymal stem cells are mainly derived from bone marrow, fetal tissue (e.g., umbilical cord and placenta), and fat. In recent years, mesenchymal stem cells derived from fetal tissue have been increasingly used because of their simple extraction.

At present, in the field of basic research and clinical application research, mesenchymal stem cells are obtained mainly by shearing fetal tissue with manual scissors so as to separate out internal stem cells. This method is time and labor intensive, and repeated shearing of tissue during the procedure can cause mechanical damage to the stem cells within, reducing the efficiency of stem cell isolation. There is still a lack of other separation means or instruments that can efficiently obtain stem cells.

Therefore, the prior art is still to be improved.

Disclosure of Invention

In view of the above-mentioned disadvantages of the prior art, an object of the present invention is to provide an automatic mesenchymal stem cell isolation apparatus, which is intended to solve the technical problems of the conventional mesenchymal stem cell isolation apparatus that much time and labor are required and the stem cell isolation efficiency is low.

The technical scheme of the invention is as follows:

an automatic mesenchymal stem cell separation device comprises a shell, a controller arranged outside the shell, a tissue separator arranged inside the shell, and a motor arranged below the tissue separator and used for driving the tissue separator to operate;

a plurality of liquid storage tanks are formed between the tissue separator and the shell through partition plates, liquid inlet pipelines and liquid outlet pipelines are arranged below the liquid storage tanks, communicating pipes are arranged in the liquid storage tanks and connected with the liquid inlet pipelines and the liquid outlet pipelines, and electromagnetic valves are arranged on the communicating pipes;

the liquid inlet pipeline and the liquid discharge pipeline are both connected with a liquid pump, and the liquid pump is connected with the tissue separator; the motor, the electromagnetic valve and the liquid pump are all electrically connected with the controller.

The automatic mesenchymal stem cell separation device is characterized in that a liquid storage bag is arranged in the liquid storage tank, and the liquid storage bag is connected with the communicating pipe through a valve connector.

The automatic mesenchymal stem cell separation device comprises a valve joint, a valve body and a valve body, wherein the valve joint comprises a pressing cap component and an elastic interface component;

the pressing cap assembly comprises a locking nut and a top pipe arranged on the locking nut;

the elastic interface assembly comprises a valve body, a supporting base plate arranged in the valve body, a stand column arranged on the supporting base plate, a spring and a circular ring piece which are sequentially sleeved on the stand column, wherein a through hole used for liquid to flow through is formed in the bottom of the stand column.

The automatic mesenchymal stem cell separation device is characterized in that the locking nut is arranged on the top pipe in a sliding manner, and the top pipe is provided with a hanging lug and a sealing ring for limiting the sliding range of the locking nut.

The automatic mesenchymal stem cell separation device comprises a cylinder body, a central shaft arranged in the cylinder body, a cutting assembly which is in threaded connection with the central shaft and can move up and down, a stirring impeller fixedly connected with the bottom end of the central shaft, and a cylinder cover arranged at the top of the cylinder body.

Automatic type mesenchymal stem cell separator, wherein, the flat axle of center pin for having symmetry cutting face and symmetry circular arc surface, cutting assembly includes blade and sword net, the blade is installed just can slide from top to bottom on the symmetry cutting face of center pin, the sword net sets up blade below and with the symmetry circular arc surface threaded connection of center pin, be provided with the couple on the central connecting portion of blade, correspond on the central connecting portion of sword net and be provided with the hook disk, the blade with the hook links to each other between the sword net.

The automatic mesenchymal stem cell separation device is characterized in that a sliding block is arranged at the edge of the knife net, and a sliding groove matched with the sliding block is formed in the inner wall of the cylinder body.

The automatic mesenchymal stem cell separation device is characterized in that the motor is connected with the central shaft of the tissue separator through a transmission gear set.

The automatic mesenchymal stem cell separation device is characterized in that a battery module for supplying power to the motor, the electromagnetic valve, the liquid pump and the controller is arranged below the liquid storage tank.

The automatic mesenchymal stem cell separation device further comprises a cart convenient for the device to move.

Has the advantages that: the invention provides an automatic mesenchymal stem cell separation device which comprises a shell, a controller arranged outside the shell, a tissue separator arranged inside the shell, and a motor used for driving the tissue separator to operate, wherein a plurality of liquid storage tanks are formed between the tissue separator and the shell, a liquid inlet pipeline and a liquid discharge pipeline are arranged below the liquid storage tanks, the liquid storage tanks are connected with the liquid inlet pipeline and the liquid discharge pipeline through a communicating pipe with an electromagnetic valve, the liquid inlet pipeline and the liquid discharge pipeline are connected with the tissue separator through a liquid pump, and the motor, the electromagnetic valve and the liquid pump are all electrically connected with the controller. The mesenchymal stem cell separation device is compact in structure, and after an operation program is set by the controller, the tissue block to be extracted can be automatically cut, sampled and extracted by only placing the tissue block to be extracted in the tissue separator, and after extraction is finished, the device can be automatically cleaned and sterilized, so that manpower is saved, and the separation efficiency of mesenchymal stem cells is improved.

Drawings

FIG. 1 is a schematic view illustrating an assembly structure of an automatic mesenchymal stem cell separation device according to the present invention;

FIG. 2 is a schematic diagram illustrating an exploded structure of an automatic mesenchymal stem cell separation device according to the present invention;

FIG. 3 is a schematic view of a valve fitting according to the present invention;

FIG. 4 is a schematic illustration of an exploded configuration of a tissue separator according to the present invention;

FIG. 5 is a schematic view of the internal structure of a tissue separator according to the present invention;

FIG. 6 is a schematic view of a blade and foil connection according to the present invention;

the reference numbers in the figures are as follows:

10-a housing; 20-a controller; 30-a tissue separator; 40-a motor; 50-a separator; 60-a liquid storage tank; 70-a liquid inlet pipeline; 80-a liquid discharge pipeline; 90-communicating tube; 100-a liquid pump; 110-a lock nut; 120-jacking pipes; 130-a valve body; 140-a support floor; 150-column; 160-a spring; 170-ring pieces; 180-through holes; 190-hanging ears; 200-sealing ring; 210-a barrel; 220-a central axis; 230-a cutting assembly; 240-stirring impeller; 250-a cartridge cover; 260-symmetrical cutting face; 270-symmetrical arc surface; 280-blade; 290-knife net; 300-hooking; 310-a hook plate; 320-a slide block; 330-a chute; 340-a drive gear set; 350-a battery module; 360-cart.

Detailed Description

The present invention provides an automatic mesenchymal stem cell separation device, and the purpose, technical scheme and effect of the present invention are more clear and definite, and the present invention is further described in detail below. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

Referring to fig. 1 and 2, the present invention provides an automatic mesenchymal stem cell separation device, comprising a housing 10, a controller 20 disposed outside the housing 10, a tissue separator 30 disposed inside the housing 10, and a motor 40 disposed below the tissue separator 30 and used for driving the tissue separator to operate;

a plurality of liquid storage tanks 60 are formed between the tissue separator 30 and the shell 10 through partition plates 50, liquid inlet pipelines 70 and liquid outlet pipelines 80 are arranged below the liquid storage tanks 60, communicating pipes 90 are arranged in the liquid storage tanks 60 and connected with the liquid inlet pipelines 70 and the liquid outlet pipelines 80, and electromagnetic valves are arranged on the communicating pipes 90;

the liquid inlet line 70 and the liquid discharge line 80 are both connected to a liquid pump 100, and the liquid pump 100 is connected to the tissue separator 30; the motor 40, the solenoid valve, and the liquid pump 100 are all electrically connected to the controller 20.

From the whole structure, the automatic mesenchymal stem cell separation device mainly comprises a control system (comprising a controller), a separation system (comprising a tissue separator and a motor), and a sample adding and cleaning system (comprising a liquid storage tank, an electromagnetic valve, a liquid inlet pipeline, a liquid discharge pipeline and a liquid pump). After an operation program is set by the controller, a tissue block to be extracted is placed in the tissue separator, alcohol and PBS buffer solution are sequentially injected through the sample adding system to sterilize and wash the tissue block, the tissue block is subjected to enzymolysis under the action of enzyme liquid after the tissue is quickly and accurately cut by the separating system, the mesenchymal stem cells are efficiently extracted, and after extraction is finished, the pipeline and the tissue separator can be cleaned and sterilized by the cleaning system.

In some embodiments, the control system in the technical scheme further has a function of automatically overhauling the running states of all the components, when some component is abnormal, a prompt is given to which component is abnormal in running (for example, no electricity is caused by damage of a battery, a pipeline does not feed liquid, a gear cannot run, and the like), the prompt can be directly displayed through an operation screen of the controller and can also be broadcasted through voice, so that the automation degree of the device is higher, the subsequent maintenance test is simpler and more convenient, in a specific application scene, before a sample is placed in the device, the device needs to automatically and preliminarily detect whether the system runs normally or not, namely, the device needs to directly run without power, after the parts in the device can normally run, the sample is placed in a prompt manner, and waste caused by the fact that the device cannot run after the sample is placed in the device can be avoided to a large extent.

In specific implementation, 1 or more communicating pipes 90 in each liquid storage tank 60 can be arranged, when 1 communicating pipe is arranged, the liquid inlet and the liquid discharge of the liquid storage tank 60 share 1 communicating pipe, and the communicating pipe has 3 ports; when being provided with 2, 1 of them links to each other with inlet line 70, and another 1 links to each other with drain line 80, all is provided with 1 solenoid valve on each communicating pipe, analogizes in proper order.

In specific implementation, 1 or more liquid pumps 100 can be provided, and when 1 liquid pump 100 is provided, the liquid pump 100 is a two-in and two-out liquid pump; when the number of the liquid pumps 100 is 2, 1 of the liquid pumps 100 is connected to the liquid inlet pipe 70, the other 1 of the liquid pumps 100 is connected to the liquid discharge pipe 80, and both the liquid pumps 100 are electrically connected to the controller 20.

In specific implementation, 4 reservoirs 60 can be provided, the liquid inlet pipe 70 and the liquid outlet pipe 80 can be provided in a circular shape, and the liquid inlet pipe 70 and the liquid outlet pipe 80 pass through the lower part of the 4 reservoirs 60.

In one embodiment, the reservoirs 60 may directly store liquids, and the liquids stored in each reservoir 60 may be the same or different, and in one embodiment, 1 reservoir may be used to store alcohol, 2 reservoirs may be used to store PBS buffer, and the other 1 reservoir may be used to store digestive enzyme solution.

In another specific embodiment, a liquid storage bag is disposed in the liquid storage tank 60, and the liquid storage bag is connected to the communication pipe 90 through a valve joint. In this kind of embodiment, be provided with the stock solution bag in reservoir 60 and store liquid, be convenient for like this on the one hand the save of liquid, can avoid liquid to volatilize and make things convenient for the dismouting simultaneously, on the other hand also can avoid the corruption of liquid to the reservoir, prevents the reduction of liquid pollution and device structural strength.

Referring to fig. 3, in a specific embodiment, the valve fitting includes a gland assembly and a resilient interface assembly;

the pressing cap assembly comprises a locking nut 110 and a top pipe 120 arranged on the locking nut 110;

the elastic interface component comprises a valve body 130, a supporting bottom plate 140 arranged in the valve body 130, a stand column 150 arranged on the supporting bottom plate 140, a spring 160 and a circular ring piece 170 which are sequentially sleeved on the stand column 150, wherein a through hole 180 for liquid to flow through is arranged at the bottom of the stand column 150. Specifically, the two ends of the spring 160 are fixedly connected to the circular ring plate 170 and the support base plate 140, respectively, and the surface of the circular ring plate 170 is flush with the surface of the valve body 130 and the surface of the pillar 150 in the initial state.

In a specific embodiment, the lock nut 110 may be fixedly connected with the top pipe 120, and in another specific embodiment, the lock nut 110 is slidably disposed on the top pipe 120, and the top pipe 120 is provided with a hanging lug 190 and a sealing ring 200 for limiting a sliding range of the lock nut 110.

In this embodiment, when the valve joint is connected, the lock nut 110 is first screwed and fixed to the valve body 130, then the top pipe 120 is pressed downward, a sealing film is arranged at a port of the top pipe 120, after the top pipe 120 contacts the circular ring piece 170, because the outer diameter of the top pipe 120 is smaller than the inner diameter of the valve body 130, and the inner diameter of the top pipe 120 is larger than the outer diameter of the upright post 150, the top pipe 120 can push the circular ring piece 170 to move downward, and meanwhile, the sealing film at the port of the top pipe 120 is pierced by the upright post 150, then the top pipe 120 continues to move downward until the sealing ring 200 on the top pipe 120 is engaged with the gap between the lock nut 110 and the top pipe 120, so that the lock of the top pipe 120 on the lock nut 110 is completed, and the liquid storage bag is connected to the communication pipe 90.

Referring to fig. 4, in a specific embodiment, the tissue separator 30 includes a cylinder 210, a central shaft 220 disposed inside the cylinder 210, a cutting assembly 230 screwed on the central shaft 220 and movable up and down, a stirring impeller 240 fixedly connected to a bottom end of the central shaft 220, and a cylinder cover 250 disposed on a top of the cylinder 210. The cylinder cover 250 is used to cover the central shaft 220, the cutting assembly 230, and the stirring impeller 240 in the cylinder 210, the stirring impeller 240 is used to stir the tissue cut by the cutting assembly 230 to mix the tissue uniformly, the bottom end of the central shaft 220 is disposed through the bottom of the cylinder 210, and the position where the central shaft 220 contacts the cylinder 210 is sealed.

Referring to fig. 5, in a specific embodiment, the central shaft 220 is a flat shaft having a symmetrical cutting surface 260 and a symmetrical arc surface 270, the cutting assembly 230 includes a blade 280 (including a central connecting portion and a cutting portion) and a blade net 290 (including a central connecting portion and a screen portion), the blade 280 is mounted on the symmetrical cutting surface 260 of the central shaft 220 and can slide up and down, the blade net 290 is disposed below the blade 280 and is in threaded connection with the symmetrical arc surface 270 of the central shaft 220, referring to fig. 6, a hook 300 is disposed on the central connecting portion of the blade 280, a hook disk 310 is correspondingly disposed on the central connecting portion of the blade net 290, and the blade 280 and the blade net 290 are hooked. The two hook discs 310 can be arranged in parallel at intervals, the hooks 300 are in a shape of '21274', the upper ends of the hooks 300 are fixedly connected with the central connecting part of the blades 280, the lower ends of the hooks 300 are inserted into the gap between the two hook discs 310, the hooks 300 can be symmetrically arranged, the blades 280 and the knife net 290 can rotate relative to each other, and the blades 280 are hooked on the blades 290, and the cutting parts of the blades 280 are obliquely arranged, so that cut tissues can quickly fall below the knife net 290 through extrusion of the cutting parts of the blades 280, and tissue cell damage caused by multiple times of cutting is avoided.

Referring to fig. 5, in a specific embodiment, a sliding block 320 is disposed at an edge of the knife net 290, and a sliding groove 330 adapted to the sliding block 320 is disposed on an inner wall of the cylinder 210. The function of the chute 330 is to limit the up-and-down movement of the blade net 290 without any rotational movement.

In a specific embodiment, 3 sliders 320 can be provided, 3 corresponding chutes 330 can be provided, and the included angle between adjacent sliders 320 is 120 °, and the advantage of providing 3 sliders 320 is to fully utilize the stability of the triangle, so that the net knife is more stable during the ascending and descending processes.

Specifically, in the operation of the tissue separator of the present invention, as the central shaft 220 rotates, the knife net 290 does not rotate upward along the central shaft 220 due to the restriction of the sliding slot 310 on the inner wall of the cylinder 210, and simultaneously pushes the blade 280 to move upward, when the knife net 290 and the blade 280 travel to the thread-free fault at the upper end of the central shaft 220, the knife net 290 stops moving upward, but the blade 280 continues to rotate along the central shaft 220 for a certain time until the remaining tissue mass at the top of the cylinder 210 is completely cut, and the blade 280 always rotates along the central shaft 220 to cut the tissue mass during the whole ascending process. The knife net 290 in the tissue separator can move upwards to compress the upper space of the cylinder 210 to extrude and fix the tissue blocks, so that the blades 280 can cut the tissue blocks more fully, the tissue separator can adapt to cutting and separation of different tissue block sizes, the cutting is more complete, manual adjustment is not needed, and the tissue separator is more convenient and fast.

Referring to FIG. 2, in one embodiment, the motor 40 is coupled to the central shaft 220 of the tissue separator 30 via a drive gear set 340. The transmission gear set 340 mainly comprises two gears with different diameters, a gear with a larger diameter is arranged on the central shaft 220, a gear with a smaller diameter is arranged on the motor 40, the transmission gear set is used for speed change, and the transmission gear set can be fixedly arranged on a support below the liquid storage tank 60.

Referring to fig. 2, in a specific embodiment, a battery module 350 is provided below the reservoir 60 for powering the motor 40, the solenoid valve, the liquid pump 100, and the controller 20. Specifically, the battery module 350 may be provided with 3 blocks, which are specifically disposed in the space below the liquid pump 100 and the transmission gear set 340, so that the device structure is more compact, and the device is powered by the battery module 350 without being connected to an external power source, and the device can be moved more flexibly.

Referring to fig. 1, in a specific embodiment, the automatic type mesenchymal stem cell separation apparatus further includes a cart 360 for facilitating movement of the apparatus. In this embodiment, the housing 10 of the device is fixedly mounted on the cart 360 and includes a cover for sealing the interior of the device.

In summary, the invention provides an automatic mesenchymal stem cell separation device, which comprises a shell, a controller arranged outside the shell, a tissue separator arranged inside the shell, and a motor for driving the tissue separator to operate, wherein a plurality of liquid storage tanks are formed between the tissue separator and the shell, a liquid inlet pipeline and a liquid discharge pipeline are arranged below the liquid storage tanks, the liquid storage tanks are connected with the liquid inlet pipeline and the liquid discharge pipeline through a communicating pipe with an electromagnetic valve, the liquid inlet pipeline and the liquid discharge pipeline are connected with the tissue separator through a liquid pump, and the motor, the electromagnetic valve and the liquid pump are all electrically connected with the controller. The mesenchymal stem cell separation device is compact in structure, and after an operation program is set by the controller, the tissue block to be extracted only needs to be placed in the tissue separator, so that the device can automatically cut and extract the tissue block in a sample adding manner, and can automatically clean and sterilize the device after extraction is finished, thereby saving manpower and improving the separation efficiency of mesenchymal stem cells.

It is to be understood that the invention is not limited to the examples described above, but that modifications and variations may be effected thereto by those of ordinary skill in the art in light of the foregoing description, and that all such modifications and variations are intended to be within the scope of the invention as defined by the appended claims.

Claims

1. An automatic mesenchymal stem cell separation device is characterized by comprising a shell, a controller arranged outside the shell, a tissue separator arranged inside the shell, and a motor arranged below the tissue separator and used for driving the tissue separator to operate;

the liquid inlet pipeline and the liquid discharge pipeline are both connected with a liquid pump, and the liquid pump is connected with the tissue separator; the motor, the electromagnetic valve and the liquid pump are all electrically connected with the controller;

the tissue separator comprises a cylinder body, a central shaft arranged in the cylinder body, a cutting assembly which is in threaded connection with the central shaft and can move up and down, a stirring impeller fixedly connected to the bottom end of the central shaft, and a cylinder cover arranged at the top of the cylinder body;

the motor is connected with the central shaft of the tissue separator through a transmission gear set;

the cutting assembly comprises a blade and a cutter mesh, the blade is mounted on the symmetrical cutting surface of the central shaft and can slide up and down, the cutter mesh is arranged below the blade and is in threaded connection with the symmetrical arc surface of the central shaft, a hook is arranged on a central connecting portion of the blade, a hook disc is correspondingly arranged on the central connecting portion of the cutter mesh, a sliding block is arranged at the edge of the cutter mesh, a sliding groove matched with the sliding block is formed in the inner wall of the barrel, and the blade and the cutter mesh are connected in a hooked mode.

2. The automatic mesenchymal stem cell separation device of claim 1, wherein a liquid storage bag is disposed in the liquid storage tank, and the liquid storage bag is connected to the communication pipe through a valve connector.

3. The automated mesenchymal stem cell separation apparatus of claim 2, wherein the valve joint comprises a press cap assembly and an elastic interface assembly;

4. The automatic mesenchymal stem cell separation device of claim 3, wherein the locking nut is slidably disposed on the top tube, and the top tube is provided with a hanging lug and a sealing ring for limiting the sliding range of the locking nut.

5. The automatic mesenchymal stem cell separation device according to claim 1 or 2, wherein a battery module for supplying power to the motor, the solenoid valve, the liquid pump and the controller is disposed below the liquid storage tank.

6. The automatic-type mesenchymal stem cell-separating device according to claim 1 or 2, further comprising a cart for facilitating movement of the device.