CN217895558U

CN217895558U - Cell separation equipment

Info

Publication number: CN217895558U
Application number: CN202221615110.5U
Authority: CN
Inventors: 翁宽; 江宏伟; 谭亮; 胡璧梁; 尹果
Original assignee: Chutian Siweikang Gene Technology Changsha Co ltd
Current assignee: Chutian Siweikang Gene Technology Changsha Co ltd
Priority date: 2022-06-24
Filing date: 2022-06-24
Publication date: 2022-11-25
Anticipated expiration: 2032-06-24

Abstract

The utility model discloses a cell separation equipment, include centrifugal cavity and be used for driving the rotatory rotary driving piece of centrifugal cavity, be equipped with joint Assembly on the centrifugal cavity, be connected with feed liquor interface, waste discharge interface and suction interface on the joint Assembly, be equipped with the water conservancy diversion seat in the centrifugal cavity, be equipped with feed liquor passageway, waste discharge passageway and suction passageway on the water conservancy diversion seat, the feed liquor interface through first connecting pipe with the feed liquor passageway intercommunication, waste discharge interface pass through the second connecting pipe with waste discharge passageway intercommunication, the suction interface pass through the third connecting pipe with suction passageway intercommunication, the suction passageway extends to centrifugal cavity's tip. A continuous separation process comprising the steps of: a) Adding a buffer solution; b) Filling liquid containing target cells; c) Layering cells; d) Separating cells; or, d') washing the target cells. The utility model has the advantages of simple and compact structure, continuous centrifugal separation and large-volume liquid treatment.

Description

Cell separation equipment

Technical Field

The utility model relates to a centrifugal equipment especially relates to a cell separation equipment.

Background

The cell therapy market is growing rapidly, the separation of cells from blood is the basic technology, and the application of automation, disposable closed systems, separation efficiency and recovery ratio are the main factors to consider. The flow channel for raw material liquid to enter the separation chamber of the existing cell separation equipment is the same as the flow channel for extracting cells from the separation chamber, so that continuous centrifugal separation cannot be realized, and meanwhile, in order to adapt to the input of large-volume raw material liquid, the volume of the separation chamber needs to be set to be larger, so that the cost is increased, and the frequency of liquid change required by a user is increased.

SUMMERY OF THE UTILITY MODEL

The to-be-solved technical problem of the utility model is to overcome the not enough of prior art, provide a simple structure, compactness, can realize continuous centrifugal separation, can handle the cell separation equipment of great volume liquid.

The utility model discloses further provide a continuous separation method of above-mentioned cell separation equipment.

In order to solve the technical problem, the utility model discloses a following technical scheme:

the utility model provides a cell separation equipment, includes centrifugal chamber and is used for driving the rotatory rotary driving piece of centrifugal chamber, be equipped with joint Assembly on the centrifugal chamber, be connected with feed liquor interface, waste discharge interface and suction interface on the joint Assembly, be equipped with the water conservancy diversion seat in the centrifugal chamber, be equipped with inlet channel, waste discharge passageway and suction passageway on the water conservancy diversion seat, the inlet channel interface through first connecting pipe with inlet channel intercommunication, waste discharge interface pass through the second connecting pipe with waste discharge passageway intercommunication, the suction interface pass through the third connecting pipe with suction passageway intercommunication, the suction passageway extends to centrifugal chamber's tip.

As a further improvement of the technical scheme: the axle center of the first connecting pipe coincides with the rotating axle center of the centrifugal chamber, the third connecting pipe is coaxially sleeved on the periphery of the first connecting pipe, a gap is formed between the first connecting pipe and the third connecting pipe, and the second connecting pipe is coaxially sleeved on the periphery of the third connecting pipe, a gap is formed between the third connecting pipe and the third connecting pipe.

As a further improvement of the above technical solution: the inlet channel is located the water conservancy diversion seat is close to one side of rotary driving piece, the channel of wasting discharge is located the water conservancy diversion seat is close to one side of joint Assembly, the suction passageway is located the inlet channel with between the channel of wasting discharge.

As a further improvement of the above technical solution: the inlet channel the waste discharge channel with the suction channel all is equipped with a plurality ofly and follows the circumference in centrifugal chamber's rotatory axle center is evenly arranged.

As a further improvement of the above technical solution: sealing elements are arranged between the first connecting pipe and the joint assembly, between the second connecting pipe and the joint assembly, and between the third connecting pipe and the joint assembly.

As a further improvement of the technical scheme: at least one side of the centrifugal chamber is provided with a conical part.

As a further improvement of the above technical solution: feed liquor interface connection has the input pump, the entry of input pump is connected with the stoste bag, trades liquid bag and reserve interface through the break-make valve respectively, be equipped with first liquid detection part on the interface of wasting discharge, the interface of wasting discharge is connected with the waste liquid bag through the break-make valve, be equipped with suction pump, second liquid detection part and colour detection part on the suction interface, the export of suction pump is connected with results bag, separation liquid bag, buffer bag and waste liquid bag through the break-make valve respectively.

A continuous separation method of the cell separation device comprises the following steps:

a) Adding a buffer solution: the suction interface fills a buffer solution into the centrifugal chamber through the third connecting pipe and the suction channel until the waste discharge interface detects that the centrifugal chamber is full of the buffer solution;

b) Filling liquid containing target cells: the liquid inlet interface fills liquid containing target cells into the centrifugal chamber through the first connecting pipe and the liquid inlet channel, the rotary driving piece drives the centrifugal chamber to rotate at the same time, the cells are gathered at the end part of the centrifugal chamber, and supernatant is discharged to the waste discharge interface through the waste discharge channel and the second connecting pipe;

c) Cell stratification: after the liquid containing the target cells is completely filled, increasing the rotating speed of the rotary driving piece to layer the cells in the centrifugal chamber;

d) Cell separation: the suction interface sucks the waste cells at the end part of the centrifugal chamber to a waste liquid bag through a third connecting pipe and a suction channel, the waste cells are switched to a harvesting bag after being sucked, the target cells are harvested until the target cells are harvested, and the buffer solution or gas is supplemented into the centrifugal chamber through the liquid inlet interface while the suction interface sucks the cells;

or, d') washing the target cells: the suction interface sucks the waste cells at the end part of the centrifugal chamber to the waste liquid bag, after the suction of the waste cells is finished, the suction interface injects a buffer solution into the centrifugal chamber through the third connecting pipe and the suction channel to elutriate target cells, and the waste discharging interface collects waste liquid.

As a further improvement of the above technical solution:

in the step b, a suction interface is used for filling the separation liquid into the centrifugal chamber;

step d' is followed by:

e) And (3) concentrating: the rotating speed of the rotary driving piece is increased to enable target cells to be gathered, the suction interface sucks waste cells in the centrifugal chamber to the harvesting bag, and the liquid inlet interface supplies buffer solution or gas to the centrifugal chamber while the suction interface sucks the target cells.

As a further improvement of the technical scheme:

and c, the waste cells are red blood cells, the target cells are white blood cells, and in the step c, the red blood cells are close to the end part of the centrifugal chamber, and the white blood cells are close to the rotating shaft center of the centrifugal chamber.

Compared with the prior art, the utility model has the advantages of: the utility model discloses a cell separation equipment, be equipped with the feed liquor interface on the joint Assembly respectively, waste discharge interface and suction interface, be equipped with inlet channel on the water conservancy diversion seat respectively, waste discharge passageway and suction passageway, the feed liquor interface, waste discharge interface and suction interface are respectively through first connecting pipe, second connecting pipe and third connecting pipe with there is inlet channel, waste discharge passageway and suction passageway intercommunication, the feed liquor in the centrifugal cavity, waste discharge and suction process can go on in step, realize continuous centrifugal separation, the liquid volume can be great relatively after the cell culture is accomplished, this cell separation equipment can adapt to the input of great volume liquid, for example 30ml volumetric centrifugal cavity can adapt to 50L's the input of target cell liquid that contains, the user can reduce centrifugal process's the liquid change.

The utility model discloses a cell separation equipment's continuous separation method has realized continuous centrifugal separation, can adapt to the input of great volume liquid, for example 30ml volumetric centrifugal chamber can adapt to 50L contain the input of target cell liquid, and the user can reduce the liquid changing of centrifugation process, can select centrifugation, washing or concentrated process in a flexible way according to user's specific demand simultaneously.

Drawings

FIG. 1 is a schematic structural view of the cell separation apparatus of the present invention.

Fig. 2 is a schematic structural view of the centrifugal chamber of the present invention.

Fig. 3 is a partially enlarged view of fig. 2.

The reference numerals in the figures denote: 1. a centrifugal chamber; 11. a quick release joint; 2. a rotary drive member; 3. a joint assembly; 4. a liquid inlet interface; 41. an input pump; 5. a waste discharge interface; 51. a first liquid detection member; 6. a suction interface; 61. a suction pump; 62. a second liquid detection component; 63. a color detection section; 7. a flow guide seat; 71. a liquid inlet channel; 72. a waste discharge channel; 73. a suction channel; 74. a suction tube; 81. a first connecting pipe; 82. a second connection pipe; 83. a third connecting pipe; 84. a seal member; 90. an on-off valve; 91. a stock solution bag; 92. a liquid change bag; 93. a standby interface; 94. harvesting the bags; 95. a separation liquid bag; 96. a buffer solution bag; 97. a waste liquid bag; 98. a bubble sensor; 100. a cell.

Detailed Description

The invention is described in further detail below with reference to the drawings and specific embodiments.

Example one

Fig. 1 to fig. 3 show the utility model discloses a cell separation equipment's an embodiment, the cell separation equipment of this embodiment, include centrifugal chamber 1 and be used for driving the rotatory rotary driving piece 2 of centrifugal chamber 1, be equipped with joint Assembly 3 on the centrifugal chamber 1, be connected with inlet port 4 on the joint Assembly 3, waste discharge interface 5 and suction interface 6, be equipped with water conservancy diversion seat 7 in the centrifugal chamber 1, be equipped with inlet channel 71 on the water conservancy diversion seat 7, waste discharge channel 72 and suction channel 73, inlet port 4 communicates with inlet channel 71 through first connecting pipe 81, waste discharge interface 5 communicates with waste discharge channel 72 through second connecting pipe 82, suction interface 6 communicates with suction channel 73 through third connecting pipe 83, suction channel 73 extends to centrifugal chamber 1's tip. Preferably, a bearing is arranged between the joint component 3 and the centrifugal chamber 1, so that when the centrifugal chamber 1 rotates, the joint component 3 can be kept not to rotate, and a pipeline connected with the joint component 3 is prevented from being wound; the centrifugal chamber 1 is mounted on the centrifugal chamber 1 through a quick-release joint 11 (such as a snap joint, a threaded joint and the like), so that the centrifugal chamber 1 can be conveniently dismounted and replaced; the rotary driving part 2 and the joint component 3 can be arranged on two sides of the centrifugal chamber 1 relatively, and the structure is compact and the layout is reasonable; the rotary drive 2 may be, for example, an electric motor or the like; a suction tube 74 may be arranged in the suction channel 73, the suction tube 74 extending to the end of the centrifugation chamber 1, although in other embodiments the suction tube 74 may also be of one-piece construction with the suction channel 73.

This cell separation equipment, be equipped with inlet port 4 on adapter module 3 respectively, waste discharge interface 5 and suction interface 6, be equipped with inlet channel 71 on water conservancy diversion seat 7 respectively, waste discharge channel 72 and suction channel 73, inlet port 4, waste discharge interface 5 and suction interface 6 are respectively through first connecting pipe 81, second connecting pipe 82 and third connecting pipe 83 with there is inlet channel 71, waste discharge channel 72 and suction channel 73 intercommunication, feed liquor in the centrifugal chamber 1, waste discharge and suction process can go on in step, realize continuous centrifugation separation, the liquid volume can be great relatively after the cell culture is accomplished, this cell separation equipment can adapt to the input of great volume liquid, for example 30ml volumetric centrifugal chamber 1 can adapt to 50L's the input of containing target cell liquid, the user can reduce the liquid change of centrifugation process.

Further, in the embodiment, the axis of the first connecting pipe 81 coincides with the rotation axis of the centrifugal chamber 1, the third connecting pipe 83 is coaxially sleeved on the periphery of the first connecting pipe 81 with a gap therebetween, and the second connecting pipe 82 is coaxially sleeved on the periphery of the third connecting pipe 83 with a gap therebetween. The liquid inlet port 4 can be directly communicated with the liquid inlet channel 71 through the first connecting pipe 81, the waste discharge port 5 can be communicated with the waste discharge channel 72 through a gap between the second connecting pipe 82 and the third connecting pipe 83, and the suction port 6 can be communicated with the suction channel 73 through a gap between the third connecting pipe 83 and the first connecting pipe 81, so that the whole structure is compact, and the occupied space is small; when the centrifugal chamber 1 rotates, the dynamic stress balance is kept, and the equipment works stably.

Furthermore, in this embodiment, the liquid inlet channel 71 is disposed on one side of the diversion seat 7 close to the rotary driving member 2, the waste discharge channel 72 is disposed on one side of the diversion seat 7 close to the joint assembly 3, and the suction channel 73 is disposed between the liquid inlet channel 71 and the waste discharge channel 72, so as to facilitate various liquids and cells to flow according to a set path, and ensure a cell separation effect.

In a preferred embodiment, the liquid inlet passage 71, the waste passage 72, and the suction passage 73 are provided in plurality and arranged uniformly in the circumferential direction of the rotation axis center of the centrifugal chamber 1. Because the axes of the first connecting pipe 81, the second connecting pipe 82 and the third connecting pipe 83 coincide with the rotation axis of the centrifugal chamber 1, the liquid inlet channel 71, the waste discharge channel 72 and the suction channel 73 are uniformly arranged around the rotation axis, which is beneficial to keeping various liquids and cells in the centrifugal chamber 1 to be uniformly distributed, and meanwhile, the liquids and cells in various directions can be discharged or sucked, so that the separation effect is ensured.

Further, the liquid inlet port 4 and the suction port 6 are disposed on one side of the joint assembly 3 away from the centrifugal chamber 1, and the waste discharge port 5 is disposed on one side of the joint assembly 3 close to the centrifugal chamber 1, which is beneficial to shortening the total length of the first connecting pipe 81, the second connecting pipe 82 and the third connecting pipe 83 and reducing the axial size of the device.

In a preferred embodiment, sealing members 84 (e.g., commonly used sealing rings) are disposed between the first connection pipe 81 and the joint assembly 3, between the second connection pipe 82 and the joint assembly 3, and between the third connection pipe 83 and the joint assembly 3, so as to maintain the sealing performance and reliability of the device and prevent leakage.

As a preferred embodiment, two opposite sides of the centrifugal chamber 1 are provided with conical parts, but in other embodiments, the centrifugal chamber 1 may also adopt a single conical structure, and a counterweight may be arranged on the side opposite to the conical part, and correspondingly, the suction channel 73 extends to the conical tip of the conical part, so that the target cells are conveniently gathered on the conical part, the suction is convenient, and the cell separation effect is ensured.

Further, in this embodiment, the liquid inlet port 4 is connected to an input pump 41, an inlet of the input pump 41 is connected to a raw liquid bag 91, a liquid changing bag 92 and a standby port 93 through an on-off valve 90 (preferably, a pinch valve, but may be other valves capable of controlling the on-off of the pipeline), the waste discharge port 5 is provided with a first liquid detecting member 51, the waste discharge port 5 is connected to a waste liquid bag 97 through the on-off valve 90, the suction port 6 is provided with a suction pump 61, a second liquid detecting member 62 and a color detecting member 63, and an outlet of the suction pump 61 is connected to a harvest bag 94, a separation liquid bag 95, a buffer liquid bag 96 and a waste liquid bag 97 through the on-off valve 90. The stock solution bag 91 is used for placing liquid containing target cells, the liquid change bag 92 can be used for placing buffer solution (such as physiological saline and the like), and the spare interface 93 can be used for connecting a gas supply part or connecting the stock solution bag 91, the liquid change bag 92 and the like in an emergency; the first liquid detection component 51 and the second liquid detection component 62 can be, for example, bubble sensors, can detect liquid in a pipeline, and detect red blood cells and white blood cells by matching with the color detection component 63, so that the running state of the device can be switched in time, and the automation and intelligence degree of the device can be improved; preferably, the spare interface 93 and each bag body are also provided with a bubble sensor; the waste liquid bag 97 is used for collecting waste liquid, waste gas cells and the like; the harvest bag 94 is used to collect the target cells, the separation fluid bag 95 is used to hold the separation fluid, and the buffer bag 96 is used to hold the buffer.

Example two

One embodiment of the continuous separation method of the cell separation apparatus described above comprises the steps of:

a) Adding a buffer solution: the suction port 6 fills the buffer solution into the centrifugal chamber 1 through the third connection pipe 83 and the suction channel 73 until the waste port 5 detects the full solution, specifically: the buffer solution in the centrifugal chamber 1 flows out to the waste discharge interface 5 through the waste discharge channel 72 and the second connecting pipe 82, and the buffer solution is detected by the first liquid detection component 51; the buffer solution is introduced from the suction interface 6, which is favorable for exhausting the air in the centrifugal chamber 1 and ensures the subsequent cell separation effect.

b) Liquid containing target cells 100 is filled: the liquid inlet interface 4 fills the liquid containing the target cells 100 into the centrifugal chamber 1 through the first connecting pipe 81 and the liquid inlet channel 71, the centrifugal chamber 1 is driven to rotate by the rotary driving member 2, the cells 100 are gathered at the end part of the centrifugal chamber 1, and the supernatant is discharged to the waste discharging interface 5 through the waste discharging channel 72 and the second connecting pipe 82;

c) Layering of cells 100: after the liquid containing the target cells 100 is completely filled, increasing the rotating speed of the rotary driving piece 2 to layer the cells 100 in the centrifugal chamber 1;

d) Cell 100 isolation: the suction interface 6 sucks the waste cells at the end part of the centrifugal chamber 1 to the waste liquid bag 97 through the third connecting pipe 83 and the suction channel 73, the waste cells are switched to the harvesting bag 94 after being sucked, and the target cells are harvested until the target cells are harvested, and the buffer solution or gas is supplemented into the centrifugal chamber 1 by the liquid inlet interface 4 while the suction interface 6 sucks the cells;

or, d') washing the target cells: the suction interface 6 sucks the waste cells at the end of the centrifugal chamber 1 to the waste liquid bag 97, after the suction of the waste cells is completed, the suction interface 6 elutriates target cells by adding buffer solution into the centrifugal chamber 1 through the third connecting pipe 83 and the suction channel 73, and the waste liquid is collected by the waste discharge interface 5.

The continuous separation method realizes continuous centrifugal separation, can adapt to the input of liquid with larger volume, for example, a centrifugal chamber 1 with 30ml volume can adapt to the input of 50L of liquid containing target cells, a user can reduce liquid change in the centrifugal process, and simultaneously, the centrifugal, washing or concentration process can be flexibly selected according to the specific requirements of the user.

Further, in the present embodiment, the first and second electrodes,

in the step b, the suction interface 6 fills the separation liquid into the centrifugal chamber 1, so that the cell separation efficiency can be effectively improved, and the loss of target cells is reduced;

step d' is followed by:

e) And (3) concentrating: the rotating speed of the rotary driving member 2 is increased to enable the target cells to be gathered, the suction interface 6 sucks the waste cells in the centrifugal chamber 1 to the harvest bag 94, the buffer solution or gas is supplemented into the centrifugal chamber 1 by the liquid inlet interface 4 while the suction interface 6 sucks the target cells, the target cells can be gathered, and the suction interface 6 can suck the target cells with higher concentration. In the concentration process, the rotating speed should be controlled to avoid cell agglomeration caused by overhigh rotating speed, which is not beneficial to cell recovery.

In a preferred embodiment, the waste cells are red blood cells, the target cells are white blood cells, and in step c, the red blood cells are near the end of the centrifugal chamber 1, and the white blood cells are near the rotation axis or the flow guide seat 7 of the centrifugal chamber 1.

Although the present invention has been described with reference to the preferred embodiments, it is not intended to be limited thereto. The technical solution of the present invention can be used by anyone skilled in the art to make many possible variations and modifications, or to modify equivalent embodiments, without departing from the scope of the technical solution of the present invention, using the technical content disclosed above. Therefore, any simple modification, equivalent change and modification made to the above embodiments by the technical essence of the present invention should fall within the protection scope of the technical solution of the present invention.

Claims

1. A cell separation apparatus comprising a centrifugation chamber (1) and a rotary drive member (2) for driving the centrifugation chamber (1) in rotation, characterized in that: be equipped with joint Assembly (3) on centrifugal chamber (1), be connected with inlet fluid interface (4), interface of wasting discharge (5) and suction interface (6) on joint Assembly (3), be equipped with water conservancy diversion seat (7) in centrifugal chamber (1), be equipped with inlet channel (71), waste discharge passageway (72) and suction channel (73) on water conservancy diversion seat (7), inlet fluid interface (4) through first connecting pipe (81) with inlet channel (71) intercommunication, waste discharge interface (5) through second connecting pipe (82) with waste discharge passageway (72) intercommunication, suction interface (6) through third connecting pipe (83) with suction channel (73) intercommunication, suction channel (73) extend to the tip of centrifugal chamber (1).

2. The cell separation apparatus according to claim 1, wherein: the axis of the first connecting pipe (81) coincides with the rotation axis of the centrifugal chamber (1), the third connecting pipe (83) is coaxially sleeved on the periphery of the first connecting pipe (81) with a gap therebetween, and the second connecting pipe (82) is coaxially sleeved on the periphery of the third connecting pipe (83) with a gap therebetween.

3. The cell separation apparatus according to claim 2, wherein: inlet channel (71) are located water conservancy diversion seat (7) are close to one side of rotary drive piece (2), waste discharge passage (72) are located water conservancy diversion seat (7) are close to one side of coupling assembling (3), suction channel (73) are located inlet channel (71) with between waste discharge passage (72).

4. The cell separation apparatus according to claim 2, wherein: liquid inlet channel (71), waste discharge channel (72) with suction channel (73) all are equipped with a plurality ofly and follow the circumference of the rotatory axle center of centrifugal chamber (1) is evenly arranged.

5. The cell separation apparatus according to claim 2, characterized in that: sealing elements (84) are arranged between the first connecting pipe (81) and the joint component (3), between the second connecting pipe (82) and the joint component (3) and between the third connecting pipe (83) and the joint component (3).

6. The cell separation apparatus according to any one of claims 1 to 5, characterized in that: at least one side of the centrifugal chamber (1) is provided with a conical part.

7. The cell separation apparatus according to any one of claims 1 to 5, characterized in that: the feed liquor interface (4) is connected with input pump (41), the entry of input pump (41) is connected with stoste bag (91), trades liquid bag (92) and reserve interface (93) through on-off valve (90) respectively, be equipped with first liquid detection part (51) on the interface of wasting discharge (5), waste discharge interface (5) are connected with waste liquid bag (97) through on-off valve (90), be equipped with suction pump (61), second liquid detection part (62) and colour detection part (63) on suction interface (6), the export of suction pump (61) is connected with results bag (94), separation liquid bag (95), buffer liquid bag (96) and waste liquid bag (97) through on-off valve (90) respectively.