TWI828453B - Gravity dispensing device, cell sub-processing machine equipped with the gravity dispensing device, and cell sub-processing method - Google Patents

Abstract

The invention relates to a gravity dispensing device, a cell sub-processing machine equipped with the gravity dispensing device, and a cell sub-processing method. The gravity dispensing device includes a funnel, a dispensing valve, a diverter and a stirring rod. The dispensing valve closes or opens the bottom opening of the funnel. The diverter is connected to the bottom of the funnel, and has an input flow channel and two output flow channels formed inside it. The input flow channel extends up and down and connects to the bottom opening of the funnel. One end of each output flow channel is connected to the input flow channel, and the other end is connected to the outside world. Stir the liquid in the funnel with a stirring rod. In this way, the cells in the liquid will not aggregate into clumps in the funnel or flow channel, which can avoid uneven cell dispensing during the cell subgeneration process, which will cause difficulties in subsequent cell preparation and quantification, and can improve the cell subgeneration process. Degree of processing automation.

Description

Gravity dispensing device, cell sub-processing machine equipped with the gravity dispensing device, and cell sub-processing method

The invention relates to a cell subgeneration device, in particular, a device that can evenly transfer liquids or cells to different containers using gravity during the cell subgeneration process.

Cell subculture is to place the cells to be cultured (such as human stem cells) in a culture container and add appropriate culture medium to allow the cells to grow and proliferate in the culture container. When the cells in the culture container proliferate to a certain density, first use enzymes and related solutions to separate the cells from the culture container, then put the separated cell solution into a centrifuge tube, and use a centrifuge to separate the cells from the solution. The cells are transferred to different containers for various subsequent operations such as subculture, cryopreservation, or bottling for shipment.

When manually loading the cell solution into centrifuge tubes, the volume or weight must be used to evenly load the cell solution into multiple centrifuge tubes to achieve relative weight balance before placing it into a centrifuge for centrifugal separation. If you want to use this cell solution Traditional manual methods are too time-consuming and complex when it comes to automating transfer processes.

In order to ensure that the cell solution can be accurately distributed into the two centrifuge tubes, a peristaltic pump is generally used for operation. Although the peristaltic pump can accurately control the flow rate, its internal pipeline length is long, and part of the absorbed cell solution will remain in the pipeline in the pump and cannot be output. Therefore, every time the pump is used, it will cause or More or less cell solution loss.

However, for cell subculture, the total amount of cell solution required for each operation is not large, and each milliliter of cell solution contains a certain number of cells. Using a peristaltic pump will cause This causes a large number of precious cells in the cell solution to be lost during the transfer process. Furthermore, when a peristaltic pump is used to transport the cell solution, the pump must pressurize the cell solution, causing the cells to rupture and be damaged due to pressure changes. Therefore, currently used peristaltic pumps result in less efficient cell subculture and increase the cost of cell preparation.

In addition, various operations still need to be performed depending on the situation before transferring the cell solution to a different container. For example, it is necessary to first confirm whether the number of cells in the culture container is sufficient; for cells with adherent growth type, special agents need to be added to separate the cells from the wall of the culture container before the transfer operation can be carried out. However, current operations are manual, which is time-consuming, difficult to control quality, and cells are easily contaminated during the process.

Therefore, the existing cell subculture equipment and methods need to be improved.

In view of the aforementioned shortcomings and shortcomings of the existing technology, when using automated equipment to transfer cell solutions, the cell solutions that have been separated from the culture containers in multiple culture containers will first be concentrated into a special container, and then a gravity dispensing device will be used. Distribute the cell solution evenly into the two centrifuge containers, and then place the two centrifuge containers symmetrically in the centrifuge for centrifugation to ensure balance during the centrifugation process. This invention provides a gravity dispensing device, a cell sub-processing machine equipped with the gravity dispensing device and a cell sub-processing method to improve the loss of cells during the transfer of the cell solution to the container.

In order to achieve the above creative purpose, the technical means used in this creation is to design a gravity dispensing device, which includes: a funnel, which has a connected internal space and a bottom opening; a dispensing valve, which is arranged on the funnel And the bottom opening can be closed or opened; a diverter piece is connected to the bottom of the funnel, and the interior of the diverter piece is formed with: An input flow channel, which extends up and down. An upper end of the input flow channel forms an input opening on the top of the diverter and is connected to the bottom opening of the funnel; two output flow channels; one end of each output flow channel is connected to the input The lower end of the flow channel and the other end of each output flow channel extend downward obliquely and are connected to the outside world; a stirring assembly has: a funnel cover that detachably covers an upper opening of the funnel; a stirring rod, It is connected to the funnel lid and used to stir the liquid inside the funnel.

In order to achieve the above creative purpose, this invention further provides a cell subculture processor, which is used to collect the cell solutions that have been separated from the cell culture boxes in a plurality of cell culture boxes into two centrifuge boxes. The cell subculture processor includes: a Base; a box processing device, which is located on the base and includes: a box positioning table, which is rotatably or movably installed on the base; a plurality of box receiving seats, each of which contains The seat is used to accommodate a cell culture box or a centrifuge box; each box holder is rotatably provided on the box positioning platform, and can be rotated to a tilting angle to pour out the cell culture box or the centrifuge box. The contents of the box; a plurality of box drive assemblies that respectively control the angles of the box seats, and each of the box drive assemblies can swing the corresponding box seat or rotate the corresponding box seat to the tilting angle; wherein, the box positioning platform can move each box holder to a first injection position; a liquid even distribution device is located on one side of the box processing device and includes: A dispensing positioning platform that can rotate or move relative to the base; at least one gravity dispensing device as mentioned above, which is installed on the dispensing positioning platform; Wherein, the dispensing positioning platform can move the at least one gravity dispensing device from below the first injection position to above the two centrifuge boxes, so that the liquid in the funnel of the at least one gravity dispensing device can be separated The two output channels flow evenly downward into the two centrifuge boxes.

In order to achieve the above-mentioned creative purpose, this invention further provides a cell subculture processing method, which is used to collect the cell solutions that have been separated from the cell culture boxes in a plurality of cell culture boxes into two centrifuge boxes. The cell subgeneration processing method includes: The above-mentioned gravity dispensing device evenly distributes the contents of the cell culture boxes into the two centrifuge boxes, and then the two centrifuge boxes are placed at opposite positions in a centrifuge device and centrifuged.

When using the cell subculture processor of this invention, first use the injection device to add the solvent that separates the cells from the cell culture box into the box, and use the box processing device to shake the cell culture box to separate the cells from the cell culture box and evenly distribute them. in the solvent, and then the box processing device rotates the cell culture box to a tilting angle, allowing the cell solution to flow from the cell culture box into the gravity dispensing device below; the cell solution in the gravity dispensing device automatically flows downwards into the diverter due to gravity. , and is evenly divided into two parts in the splitter, and enters the two centrifuge boxes from the two output flow channels.

The advantages of this creation are as follows:

First, the diverter of the gravity dispensing device is arranged below the funnel, and the input flow channel extends up and down. At the same time, the output flow channel extends obliquely downward. Therefore, the liquid in the funnel will automatically flow from the funnel through the input flow channel due to gravity. and output flow channel and flow into the centrifuge box below, without remaining in the funnel, input flow channel or output flow channel, which can prevent cells from being lost during the transfer process, and can also avoid cells from being ruptured and damaged due to pump pressure. It also effectively concentrates the cell solutions that have been separated from the cell culture boxes in multiple cell culture boxes and then evenly distributes them to the two centrifuge boxes. The program is automated, thereby improving the efficiency of cell subculture processing, improving the cell survival yield, and reducing the cell Cost of subculture.

Second, the stirring rod of the gravity dispensing device can continuously stir the cell solution in the funnel, thereby preventing the cells in the solution from aggregating into clusters or adhering to the surface of the funnel or flow channel, causing a difference in the flow rate of the two output flow channels, ensuring that The cell solution was evenly distributed into the two centrifuge boxes.

Third, the cell subculture processing machine can replace traditional manual work. It automatically shakes the cell culture box and pours the cell solution into the gravity dispensing device. The gravity dispensing device evenly distributes the cell solution into the two centrifuge boxes, which can reduce manual work. It has the advantages of high efficiency, stable quality and avoids contamination in the cell subculture process.

Furthermore, in the gravity dispensing device, at least one auxiliary channel is formed inside the diverting member, one end of which is connected to one of the output flow channels, and the other end extends upward obliquely and is connected to the outside world.

Furthermore, in the gravity dispensing device, the number of the at least one auxiliary channel of the splitting member is two, and the two auxiliary channels are respectively connected to the two output flow channels, and are respectively along the same straight line with the two output flow channels. extend.

Furthermore, in the gravity dispensing device, the dispensing valve includes a first ferromagnetic component, which is movably disposed in the internal space of the funnel and is driven by its own weight toward the funnel. The bottom opening moves and closes the bottom opening; a valve actuator is located outside the funnel, and a moving end of the valve actuator can move toward the first ferromagnetic component to an open position; a second ferromagnetic component, which is located at the moving end of the valve actuator; the second ferromagnetic component and the first ferromagnetic component magnetically attract or repel each other; wherein, when the moving end of the valve actuator is located at the In the open position, the first ferromagnetic component is driven by the second ferromagnetic component to open the bottom opening.

Furthermore, the cell subculture processor, wherein: the box positioning platform can move each box holder to a second injection position; the cell subculture processor has a medicine injection device; The medicine injection device is provided on the base and includes a plurality of liquid injection heads; each liquid injection head is used to connect a medicine, and And it can be moved above the second injection position to inject the connected medicament into the cell culture box located on the box holder of the second injection position.

Furthermore, in the cell subculture processor, the box positioning platform of the box processing device can move each box holder to a box receiving position or a box output position; The processor includes: a main box moving device, which is used to move the two centrifuge boxes located below at least one gravity dispensing device to the box receptacle located at the box receiving position; a centrifugal device, It is provided on one side of the box processing device; an auxiliary box moving device is used to move the centrifuge box in the box receptacle at the box output position into the centrifuge device, and is used to Move the centrifuge box in the centrifuge device to the box holder at the box output position.

Furthermore, the cell subculture processor, wherein: the box positioning platform of the box processing device can move each box holder to a box output position; the cell subculture processor includes: An optical detection device is provided on one side of the box processing device and is used to detect the number of cells in the cell culture boxes; an auxiliary box moving device is used to move the box at the output position of the box. The cell culture box in the box receptacle moves into the optical detection device, and is used to move the cell culture box in the optical detection device to the box receptacle at the box output position.

Furthermore, in the cell subculture processor, the box positioning platform of the box processing device is rotatably mounted on the base, and each box can be accommodated in the box. position, the output position of the box, the first injection position and a second injection position; the cell sub-processing machine has an injection device, which is provided on the base and includes a plurality of injection devices heads; each injection head is used to connect a medicament, and can be moved above the second injection position to inject the connected medicament into the cell culture on the box holder located at the second injection position. Inside the box; the cell subculture processor includes an optical detection device located on one side of the box processing device; the auxiliary box moving device is used to move the cells in the box receptacle at the output position of the box The cell culture box moves into the optical detection device, and is used to move the cell culture box in the optical detection device to the box output position in the box holder; wherein, the second injection position and the box output position are respectively located opposite to the box processing device; the centrifugal device, the auxiliary box moving device and the optical detection device are located in the The side of the box processing device facing the box output position, and the medicine injection device are located on the side of the box processing device facing the second medicine injection position.

Furthermore, the cell subculture processor, wherein: the box positioning platform of the box processing device can move each box holder to a box receiving position; the cell subculture processor includes a The main box moving device is used to move the empty cell culture box located in the box receiving position at the box receiving position to below the at least one gravity dispensing device, so that the at least one gravity dispensing device The cell solution in the funnel of the device can flow into the cell culture box for subculture of cells.

10: base

20:Box processing device

21: Box positioning table

211: Box receiving position

212: Box output position

213: First injection position

214: Second injection position

22: Box holder

221:Shaft

222: driven wheel

23: Box drive assembly

231: Linear module

232:Motor

233:Driving wheel

30: Injection device

31: Liquid injection head

40: Liquid average distribution device

41:Dispensing positioning table

42: Gravity dispensing device

421:Funnel

4211:Flow channel

422:Dispensing valve

4221: The first ferromagnetic part

4222:Valve actuator

4222A:Mobile terminal

4223: Second ferromagnetic part

423: Diversion piece

4231:Input flow channel

4232:Output runner

4233: Auxiliary channel

424:Funnel cover

425: Stirring component

4251: Linear module

4251A:Sliding seat

4252: Stirring motor

4253:Stirring stick

50: Main box moving device

51: The first multi-axis transfer mechanism

511: Linear module

512:Clamp

52: Box temporary storage rack

53:First conveyor

54:Second conveyor

55:Third conveyor

56: The second multi-axis transfer mechanism

57: Bottle transfer mechanism

58: Cell medicine bottle cap clamping mechanism

60: Auxiliary box moving device

70:Centrifugal device

80: Optical detection device

81: Bottle cap mechanism

91: Culture box

92:Centrifuge box

Figure 1 is a three-dimensional appearance view of the cell subculture processor of this invention.

Figure 2 is a three-dimensional appearance view of the internal structure of the cell subculture processor of this invention.

Figure 3 is a schematic top view of the internal structure of the cell subculture processor of this invention.

Figure 4 is a simplified top view of the internal structure of the cell subculture processor of this invention.

Figure 5 is a three-dimensional component exploded view of the connection between the box processing device and the main box moving device of the cell subculture processor of the present invention.

Figure 6 is a three-dimensional appearance view of the box processing device of the cell subculture processor of the present invention.

Figure 7 is a partial three-dimensional component exploded view of the box processing device for cell subculture processing of the present invention.

Figures 8 to 10 are schematic side views of the cell subculture processor of the present invention, showing that the first multi-axis transfer mechanism of the main box moving device moves a cell culture box to the box processing device.

Figure 11 is a schematic side view of the cell subculture processor of this invention, showing that the bottle cap mechanism descends and clamps the upper cover of the cell culture box.

Figure 12 is a schematic side view of the cell subculture processor of this invention, showing that the liquid injection head of the medicine injection device is aligned with the cell culture box.

Figure 13 is a schematic side view of the cell subculture processor of the present invention, showing that the box processing device shakes the cell culture box.

Figures 14 to 16 are schematic top views of the cell subculture processor of this invention, showing that the auxiliary box moving device can move between the box output position, the centrifugal device, and the optical detection device.

Figure 17 is a partial three-dimensional component exploded view of the liquid equal distribution device of the cell subculture processor of the present invention.

Figure 18 is a three-dimensional component exploded view of the liquid equal distribution device of the cell subculture processor of the present invention.

Figure 19 is a partial three-dimensional component exploded view of the gravity dispensing device of the cell subculture processor of this invention.

Figure 20 is a partial cross-sectional schematic diagram of the gravity dispensing device of the cell subculture processor of this invention, showing that the first ferromagnetic part closes the bottom opening of the funnel.

Figures 21 and 22 are schematic side views of the box processing device of the cell subculture processor of the present invention, showing that the cell culture box is rotated to pour the contents into the gravity dispensing device below.

Figures 23 and 24 are schematic side views of the gravity dispensing device of the cell subculture processor of the present invention, showing that the stirring motor is lowered and connected to the stirring rod.

Figure 25 is a schematic side view of the gravity dispensing device of the cell subculture processor of this invention, showing the dispensing valve opening the bottom opening of the funnel.

Figure 26 is a partial three-dimensional component exploded view of the cell subculture processor of this invention.

Figure 27 is a partial side view of the cell subculture processor of this invention.

Figure 28 is a partial cross-sectional schematic diagram of the cell subculture processor of this invention, showing that the contents of the gravity dispensing device flow evenly into the two centrifuge boxes below.

Please refer to Figures 1 to 4 and 28. The cell subculture processor of the present invention is used to first concentrate the contents (i.e., cell solutions) in a plurality of culture boxes 91 and then evenly distribute them into two centrifuge boxes 92 for processing. Centrifugal separation, the cells obtained after centrifugation can continue to be used for subculture in other cell culture equipment. The cell sub-processing machine can preferably also transfer the separated cells to cryovials (not shown in the figure) for cryopreservation or transfer the separated cells to cell medicine containers (not shown in the figure) for direct shipment, depending on the situation. .

The cell culture box 91 and the centrifuge box 92 are preferably boxes with the same shape, and the cell culture box 91 used in this embodiment can be directly placed in a centrifuge for centrifugal separation. Therefore, the centrifuge box 92 can also be of multiple shapes. Two of the 91 cell culture boxes.

The processor of the present invention includes a base 10, a box processing device 20 and a liquid even distribution device 40; and in this embodiment, it further includes a medicine injection device 30, a main box moving device 50, an auxiliary A box moving device 60, a centrifugal device 70 and an optical detection device 80.

Please refer to FIGS. 4 to 7 . The aforementioned box processing device 20 is located on the base 10 and includes a box positioning platform 21 , a plurality of box receiving seats 22 and a plurality of box driving assemblies 23 .

The box positioning table 21 is rotatably disposed on the base 10, and is preferably an indexing plate in the prior art. The box positioning table 21 rotates 90 degrees each time, thereby defining four fixed positions in the space, which are respectively a box receiving position 211, a box output position 212, a first injection position 213 and A second injection position 214. The second medicine injection position 214 and the box output position 212 are respectively located opposite to the box processing device 20 . In terms commonly used in this field, the box positioning station 21 has four stations. The aforementioned box receiving position 211, box output position 212, first injection position 213 and second injection position 214 are the box positioning positions. There are four workstations on stage 21.

The number of box receptacles 22 is preferably four, and they are arranged at 90-degree intervals on the box positioning platform 21. The box positioning platform 21 can position each box receptacle 22 at the box receiving position 211 and the box output. Location 212. Circular movement between the first injection position 213 and the second injection position 214. The number of box holders 22 is not limited to four, and may also be one or a plurality other than four.

The action mode of the box positioning platform 21 is not limited to the above. In other preferred embodiments, the box positioning platform 21 can be designed to move the box holder 22 between multiple specific positions in a linear or turning manner. In addition, the box positioning platform 21 only needs to be able to move each box receptacle 22 to the first injection position 213 .

Please refer to FIG. 7 , FIG. 21 and FIG. 22 . Each box housing 22 can accommodate a cell culture box 91 or a centrifuge box 92 . Each box holder 22 can be rotatably disposed on the box positioning platform 21 with a shaft 221 as an axis, and can be rotated to a tilting angle (as shown in FIG. 22 ) to pour out the contents of the cell culture box 91 or the centrifuge box 92 . The shaft 221 is preferably arranged horizontally.

Please refer to FIG. 6 , FIG. 7 and FIG. 13 . The number of box driving assemblies 23 is the same as that of the box receiving seats 22 . The multiple box driving assemblies 23 control the angles of the multiple box receiving seats 22 respectively. Each box driving assembly 23 can swing the corresponding box holder 22 or rotate the corresponding box holder 22 to a tilting angle.

In this embodiment, each box driving component 23 is disposed on the base 10 and includes a linear module 231, a motor 232 and at least one driving wheel 233. The linear module 231 is disposed on the base 10, and the motor 232 and the driving wheel 233 are disposed on a sliding seat of the linear module 231. The sliding seat can be controlled to move toward the box receptacle 22, so that the driving wheel 233 abuts against the A driven wheel 222 is connected to the shaft 221. At this time, the motor 232 of the box driving assembly 23 can control the angle of the corresponding box receptacle 22; the linear module 231 can also be oriented away from the box receptacle 22. direction movement. By arranging the box driving assembly 23 on the base 10 instead of the box positioning table 21, the wiring complexity of the linear module 231 and the motor 232 can be simplified.

Please refer to FIG. 3 , FIG. 4 and FIG. 12 . The aforementioned injection device 30 is provided on the base 10 and includes a plurality of injection heads 31 . Each liquid injection head 31 is connected to a medicine, and can be moved above the second medicine injection position 214 to inject the connected medicine into the fine liquid on the box receptacle 22 of the second medicine injection position 214 . inside the cell culture box 91. The injection device 30 is preferably located on the side of the box processing device 20 facing the second injection position 214 .

In this embodiment, the medicine injection device 30 is a turret. A plurality of liquid injection heads 31 are arranged around the medicine injection device 30 , and a specific liquid injection head 31 can be rotated to above the second medicine injection position 214 for injection. Medicine. The medicine connected to the injection head 31 includes a separation medicine that can separate the adherent growth cells from the wall of the cell culture box 91 and a neutralizing medicine that can neutralize the separation medicine.

Please refer to FIG. 3 , FIG. 4 , FIG. 17 and FIG. 18 . The aforementioned liquid even distribution device 40 is located on one side of the box processing device 20 and includes a dispensing positioning table 41 and at least one gravity dispensing device. 42. The dispensing positioning table 41 can rotate relative to the base 10, and is preferably an indexing plate in the prior art. The height of the dispensing positioning platform 41 is lower than the box positioning platform 21 but higher than the second centrifuge box 92 . One of the workstations on the dispensing positioning table 41 is located below the first injection position 213 of the box positioning table 21 , and the other workstation is located above the second centrifuge box 92 .

Please refer to FIGS. 18 to 20 . The number of gravity dispensing devices 42 is preferably four, and they are arranged on the dispensing positioning platform 41 at surrounding intervals. Each gravity dispensing device includes a funnel 421, a dispensing valve 422, a diverter 423 and a stirring assembly 425, and preferably has a funnel cover 424.

The funnel 421 has a connected inner space and a bottom opening. A partial dispensing valve 422 (ie, the first ferromagnetic component 4221 described below) is disposed in the funnel 421 and can close or open the bottom opening of the funnel 421. The fact that the dispensing valve 422 closes the bottom opening of the funnel 421 means that the dispensing valve 422 prevents the liquid inside the funnel 421 from flowing out from the bottom opening, and is not limited to directly blocking the bottom opening.

In this embodiment, a flow channel 4211 (as shown in FIG. 20 ) is formed in the internal space of the funnel 421 at the bottom of the funnel 421 . The flow channel 4211 extends up and down, and the lower end of the flow channel 4211 forms the aforementioned bottom opening. A first ferromagnetic component 4221 of the dispensing valve 422 is disposed in the bottom flow channel 4211 and is controlled to block or open the flow channel 4211, thereby closing or opening the bottom opening of the funnel 421.

Please refer to FIG. 20 and FIG. 23 to FIG. 25 . The dispensing valve 422 includes the aforementioned first ferromagnetic component 4221 , a valve actuator 4222 and a second ferromagnetic component 4223 . The first ferromagnetic member 4221 is movably disposed in the bottom flow channel 4211 of the funnel 421, and is driven by its own weight to move toward the bottom opening of the funnel 421 and close the bottom opening (as shown in Figure 20). The first ferromagnetic component 4221 is preferably a sphere with a metal core.

The valve actuator 4222 is disposed outside the funnel 421; a moving end 4222A of the valve actuator 4222 can be controlled to move toward the first ferromagnetic component 4221 to an open position (as shown in Figure 25). Specifically, the valve actuator 4222 is a cylinder, which is disposed on the base 10 and can be controlled by air pressure to move a slide block of the valve actuator 4222 toward the first ferromagnetic component 4221 .

The second ferromagnetic component 4223 is provided at the moving end 4222A of the valve actuator 4222. The second ferromagnetic part 4223 and the first ferromagnetic part 4221 magnetically attract or repel each other. When the moving end 4222A of the valve actuator 4222 is in the open position, the first ferromagnetic part 4221 is driven by the second ferromagnetic part 4223 And open the bottom opening of funnel 421.

In this embodiment, the second ferromagnetic component 4223 is a magnet. When the moving end 4222A of the valve actuator 4222 is in the open position, the first ferromagnetic component 4221 is attracted by the second ferromagnetic component 4223 to open the flow channel 4211 . In other preferred embodiments, the first ferromagnetic component 4221 and the second ferromagnetic component 4223 can also be two magnets that attract or repel each other, or the second ferromagnetic component 4223 can also generate magnetic force after being energized. One coil.

Please refer to FIGS. 18 to 20 . The diverter 423 is connected to the bottom of the funnel 421 . An input channel 4231 and two output channels 4232 are formed inside the diverter 423 , and preferably two auxiliary channels 4233 are formed therein. The input flow channel 4231 extends up and down. An upper end of the input flow channel 4231 forms an input opening on the top of the diverter 423 and is connected to the bottom opening of the funnel 421 . One end of each output flow channel 4232 is connected to the lower end of the input flow channel 4231, and the other end of each output flow channel 4232 extends downward obliquely and is connected to the outside world. The liquid flows into the two output flow channels 4232 through an input flow channel 4231, so that the first input flow channel 4231 The liquid inside flows out of the diverter 423 equally from the two output channels 4232 respectively. In this embodiment, each output flow channel 4232 is linear, and its lower end is not directly connected to the outside world, but is connected to the outside world through a vertical flow channel.

One end of each auxiliary channel 4233 is connected to the output flow channel 4232, and the other end extends upward obliquely and is connected to the outside world. Specifically, the two auxiliary channels 4233 extend along the same straight line as the two output flow channels 4232 respectively. Thereby, the auxiliary channel 4233 can balance the pressure in the two output channels 4232, thereby stabilizing the liquid flow rate of the two output channels 4232, and further ensuring that the fluid in the input channel 4231 flows out of the splitter 423 from the two output channels 4232 equally. . In addition, designing each auxiliary channel 4233 and one of the output flow channels 4232 to extend along the same straight line also makes the output flow channel 4232 easy to clean.

Please refer to FIGS. 18 , 20 , 23 and 24 . The funnel cover 424 detachably covers an upper opening of the funnel 421 . The stirring component 425 has a linear module 4251, a stirring motor 4252 and a stirring rod 4253. The linear module 4251 is disposed on the base 10 and can be controlled to move a sliding base 4251A thereon. The stirring motor 4252 is arranged on the sliding seat 4251A of the linear module 4251. The stirring rod 4253 rotatably penetrates the funnel cover 424 and can stir the liquid in the funnel 421.

When the funnel cover 424 covers the funnel 421, the linear module 4251 can move the stirring motor 4252 in the direction of the stirring rod 4253 (as shown in Figure 24), so that the output shaft of the stirring motor 4252 engages the upper end of the stirring rod 4253. At this time The stirring motor 4252 can drive the stirring rod 4253 to rotate.

The specific structure of the stirring assembly 425 is not limited to the above, as long as it has a stirring rod 4253 and can stir the liquid in the funnel 421 with the stirring rod 4253.

Please refer to Figure 3, Figure 4, Figure 18 and Figure 20. The rotation of the aforementioned dispensing positioning table 41 can move each gravity dispensing device 42 from the bottom of the first injection position 213 to the top of the second centrifuge box 92. upward, so that the liquid in the funnel of each gravity dispensing device 42 can flow downward into the two centrifuge boxes 92 through the two output flow channels 4232 of the diverter 423 respectively. In this embodiment, in addition to rotating on the horizontal plane, the dispensing positioning table 41 can also move linearly on the horizontal plane close to or away from the cartridge processing device 20. The positioning table 41 moves the gravity dispensing device 42 from below the first medicine injection position 213 to above the second centrifuge box 92 through a combination of rotation and linear movement.

The action mode of the dispensing positioning table 41 is not limited to rotation. In other preferred embodiments, the dispensing positioning platform 41 can be designed to move the gravity dispensing device 42 between multiple specific positions in a linear or turning manner.

Please refer to FIGS. 2 to 4 and 28 . The aforementioned main box moving device 50 can move the two centrifuge boxes 92 located below the gravity dispensing device 42 to the box receiving position 211 at the box receiving position. Within 22. In addition, the main box moving device 50 can also move the cell culture box 91 located in the box receiving position 211 to below the gravity dispensing device 42, so that the cell culture box 91 in the funnel 421 of the gravity dispensing device 42 can be moved. The liquid can flow into the cell culture box 91, whereby the present invention can recycle two of the plurality of cell culture boxes 91 and use them as centrifuge boxes 92.

Specifically, the main box moving device 50 includes a first multi-axis transfer mechanism 51, a box temporary storage rack 52, a first conveyor 53, a second conveyor 54, a third conveyor 55, A second multi-axis transfer mechanism 56 and a bottling transfer mechanism 57.

Please refer to FIG. 5 and FIG. 8 to FIG. 10 . The first multi-axis transfer mechanism 51 is preferably a three-axis transfer mechanism with three linear modules 511 , in which the linear modules 511 in the vertical direction are A clamping claw 512 is provided at the lower end, and the clamping claw 512 can clamp the box body 22 located at the box receiving position 211, the box temporary storage rack 52, or the box body (i.e., the cell culture box 91 or the centrifuge box 91) in the first conveyor 53. Box 92), and the clamped box can also be placed in any of the above three. Furthermore, the clamping jaw 512 can rotate 90 degrees along a horizontal axis to change the angle of the box being clamped.

Twelve boxes can be placed in the box temporary storage rack 52 to increase the flexibility of use. The first conveyor 53 is equipped with a translation and lifting mechanism, which can move the box from the bottom to the top for the first multi-axis transfer mechanism 51 to pick up or place. The second conveyor 54 and the third conveyor 55 are both linear conveyor belts, one end of which penetrates into the bottom of the box temporary storage rack 52 and the other end extends to the second multi-axis transfer mechanism 56 . The second multi-axis transfer mechanism 56 It is a three-axis transfer mechanism with three linear modules 511, which can pick up and place boxes between the second conveyor 54, the third conveyor 55 and the bottling transfer mechanism 57.

Please refer to Figures 26 to 28. The bottling transfer mechanism 57 can simultaneously hold two centrifuge boxes 92, two cell culture boxes 91, two freezing tubes (not shown in the figure) or two shipping containers (in the figure). (not shown), and the opening of the clamped container can be aligned with the two output channels 4232 of one of the gravity dispensing devices 42, so that the liquid in the gravity dispensing device 42 flows to the clamped container. in two containers. In addition, the bottling transfer mechanism 57 can also move the filled container out from below the box positioning table 21 for the second multi-axis transfer mechanism 56 to pick up.

In this embodiment, a cell medicine bottle cap clamping mechanism 58 is provided below the second multi-axis transfer mechanism 56, which can operate the cap of the cell medicine bottle in cooperation with the second multi-axis transfer mechanism 56.

Please refer to FIGS. 2 to 4 . The aforementioned auxiliary box moving device 60 , centrifugal device 70 and optical detection device 80 are disposed on the side of the box processing device 20 facing the box output position 212 . The centrifuge device 70 and the optical detection device 80 are standard components in the prior art. The optical detection device 80 can detect the number of cells in the cell culture box 91 to determine whether the cultured cells meet quality requirements.

Please refer to FIGS. 14 to 16 . The auxiliary box moving device 60 can move the centrifuge box 92 or cell culture box 91 located in the box receptacle 22 at the box output position 212 into the centrifuge device 70 , and can Move the centrifuge box 92 or cell culture box 91 in the centrifuge device 70 to the box receptacle 22 at the box output position 212 . In addition, the auxiliary box moving device 60 can also move the centrifuge box 92 or the cell culture box 91 located in the box receptacle 22 at the box output position 212 to the optical detection device 80 , and can also move the centrifuge box 92 or the cell culture box 91 in the optical detection device 80 The centrifuge box 92 or cell culture box 91 moves to the box receptacle 22 at the box output position 212 . The auxiliary box moving device 60 is preferably a robotic arm.

The cell sub-processing method of this invention is preferably carried out with the above-mentioned cell sub-processing machine. The cell sub-processing machine can automatically perform a variety of different cell sub-processing methods in cell culture. Method, the following takes the passage process of adherent growth cells as an example to illustrate one of the cell passage processing methods.

Please refer to FIGS. 3 to 5 . First, a plurality of cell culture boxes 91 that have been detected to be full of cells will be transported by an external mechanism to the first conveyor 53 of the main box moving device 50 . Next, please refer to FIGS. 8 to 10 . The first multi-axis transfer mechanism 51 clamps the cell culture box 91 in the first conveyor 53 and places the cell culture box 91 into the box located at the box receiving position 211 . In the body holder 22, during the process, the clamping claws 512 of the first multi-axis transfer mechanism 51 will change the cell culture box 91 from a horizontal posture to a vertical posture (as shown in Figures 8 to 9) to facilitate subsequent operations.

Please refer to Figure 4 and Figure 11. Then, the cell culture box 91 is moved from the box receiving position 211 to the second injection position 214. At this time, the bottle cap mechanism 81 will descend and clamp the cell culture box 91. Cover.

Please refer to Figure 12. Next, the bottle cap mechanism 81 unscrews the upper cover of the cell culture box 91 and moves it to one side. The box driving assembly 23 rotates the box receptacle 22 to pour out the contents of the cell culture box 91. , then move an injection head 31 of the injection device 30 above the second injection position 214 and inject a separation agent that can separate the adherent growth cells from the wall of the cell culture box 91 .

Please refer to Figure 4 and Figure 13. Then, after the bottle cap mechanism 81 closes the upper cover of the cell culture box 91, the cell culture box 91 is moved to the first injection position 213, and the box driving assembly 23 swings for cell culture. Box 91 to separate the cells from the wall.

Please refer to Figures 14 and 15. Next, the cell culture box 91 is moved to the box output position 212, and then the auxiliary box moving device 60 picks up the cell culture box 91 and places it into the optical detection device 80 for judgment. Whether the number of cells is sufficient. After the determination is completed, the cell culture box 91 is moved back to the box output position 212 and then moved to the second injection position 214 . At the second injection position 214, the injection device 30 uses another injection head 31 to add the neutralizing agent into the cell culture box 91 to stop the action of the separation agent.

Please refer to FIG. 4 , FIG. 21 and FIG. 22 . Then, the cell culture box 91 is moved to the first injection position 213 , and the box driving assembly 23 rotates the box receptacle 22 to inject the contents of the cell culture box 91 Pour into the funnel 421 of the gravity dispensing device 42 below. Before performing this step, the funnel cover 424 above the funnel 421 has been removed. Then, repeat the above steps for the remaining cell culture boxes 91 to collect the cell solutions of all the cell culture boxes 91 into the funnel 421 .

Among all the above-mentioned cell culture boxes 91, after the contents of the first two cell culture boxes 91 are poured, the cell culture box 91 will be moved to the box receiving position 211, and then through the first multi-axis of the main box moving device 50 The transfer mechanism 51 , the first conveyor 53 , the second conveyor 54 , the second multi-axis transfer mechanism 56 and the bottling transfer mechanism 57 move the lower part of one of the gravity dispensing devices 42 to be used as the centrifuge box 92 .

Please refer to Figures 23 and 24. After all the cell solutions in the cell culture boxes 91 are concentrated into the funnel 421, the dispensing positioning table 41 moves the gravity dispensing device 42 clockwise to the top of the two centrifuge boxes 92, and at the same time the stirring assembly 425 The sliding seat 4251A descends, so that the stirring motor 4252 is combined with the upper end of the stirring rod 4253 and continues to stir the cell solution in the funnel 421.

Please refer to Figure 25 and Figure 28. Then, the valve actuator 4222 of the dispensing valve 422 moves the second ferromagnetic component 4223 toward the first ferromagnetic component 4221 to the open position, so that the cells in the funnel 421 The solution is evenly filled into the two centrifuge boxes 92 .

Please refer to Figure 4. Finally, the two centrifuge boxes 92 are moved to the box processing device 20 by the main box moving device 50, and then moved to the opposite place in the centrifuge device 70 by the auxiliary box moving device 60 for centrifugal separation. After centrifugation is completed, subsequent cell subculture operations can be continued.

The cell subculture processing method of the present invention is not limited to the above, as long as the gravity dispensing device 42 is used to evenly distribute the contents of multiple cell culture boxes 91 into two centrifuge boxes 92, and then the two centrifuge boxes 92 are placed separately. It is sufficient to perform centrifugal separation at the opposite position in a centrifugal device 70 . In addition to automatically executing the passage process of adherent growing cells, the cell subculture processor can also automatically execute the primary passage process. Sample cell extraction process, cell culture box medium replacement process, cell culture box optical detection process, part of the cell cryopreservation process such as cell freezing tube packaging, part of the cell thawing process, fine package pharmaceutical bottling and shipping process, etc. Prepare related processes.

To sum up, in this invention, the diverter 423 of the gravity dispensing device 42 is arranged below the funnel 421, the input flow channel 4231 extends up and down, and the output flow channel 4232 extends obliquely downward, whereby the liquid in the funnel 421 will Due to gravity, it automatically flows from the funnel 421 through the input flow channel 4231 and the output flow channel 4232 into the centrifuge box 92 below. It will not remain in the funnel 421, the input flow channel 4231 or the output flow channel 4232, which can prevent cells from transferring. Lost or damaged during the process, the cell solution that has been separated from the cell culture boxes in the plurality of cell culture boxes 91 is effectively centralized and then evenly distributed to the two centrifuge boxes 92, thereby automating the process, thereby improving the efficiency of cell subculture processing and reducing the cost of cell subculture. The cost of generation culture.

Furthermore, the stirring rod 4253 of the gravity dispensing device 42 can continuously stir the cell solution in the funnel 421, thereby preventing the cells in the cell solution from aggregating into clumps or adhering to the surface of the funnel 421 or the flow channel, thereby causing the two output flow channels to The difference in flow rate of 4232 can ensure that the cell solution is evenly distributed into the two centrifuge boxes 92.

Furthermore, the cell subculture processing function can replace traditional manual work, automatically shake the cell culture box 91 and pour the contents into the gravity dispensing device 42, and use the gravity dispensing device 42 to evenly distribute the liquid into the two centrifuge boxes 92 , can reduce the degree of manual work, has the advantages of high efficiency, stable quality and avoids cell contamination.

The above descriptions are only preferred embodiments of the present invention, and do not limit the present invention in any form. Although the present invention has been disclosed as above in terms of preferred embodiments, it is not intended to limit the present invention. Any technical field that has A person with ordinary knowledge can use the technical content disclosed above to make some changes or modify it into equivalent embodiments with equivalent changes without departing from the scope of the technical solution of this invention. The technical essence of creation is crucial to the implementation of the above Any simple modifications, equivalent changes and modifications made in the example still fall within the scope of this creative technical solution.

20:Box processing device

22: Box holder

30: Injection device

31: Liquid injection head

40: Liquid average distribution device

50: Main box moving device

51: The first multi-axis transfer mechanism

52: Box temporary storage rack

53:First conveyor

56: The second multi-axis transfer mechanism

57: Bottle transfer mechanism

60: Auxiliary box moving device

70:Centrifugal device

80: Optical detection device

91: Culture box

Claims (11)

A gravity dispensing device, including: a funnel, which has a connected internal space and a bottom opening; a dispensing valve, which is provided on the funnel and can close or open the bottom opening; a diverter, which connects the The bottom of the funnel, the inside of the diverter is formed with: an input flow channel, which extends up and down, an upper end of the input flow channel forms an input opening at the top of the diverter and is connected to the bottom opening of the funnel; two output flows channel; one end of each output channel is connected to the lower end of the input channel, and the other end of each output channel extends downward obliquely and is connected to the outside world; a stirring assembly has: a funnel cover, which can be detachably covered An upper opening of the funnel is provided; a stirring rod is connected to the funnel cover and used to stir the liquid in the funnel. The gravity dispensing device according to claim 1, wherein at least one auxiliary channel is formed inside the diverting member, one end of which is connected to one of the output flow channels, and the other end extends upward obliquely and is connected to the outside world. The gravity dispensing device as described in claim 2, wherein the number of the at least one auxiliary channel of the splitting member is two, and the two auxiliary channels are respectively connected to the two output flow channels, and are respectively along the same straight line with the two output flow channels. extend. The gravity dispensing device according to any one of claims 1 to 3, wherein the dispensing valve includes: a first ferromagnetic component, which is movably disposed in the internal space of the funnel and is moved by its own weight. Driven to move toward the bottom opening of the funnel and close the bottom opening; a valve actuator located outside the funnel, a moving end of the valve actuator can move toward the first ferromagnetic component to an opening Location; A second ferromagnetic component is provided at the moving end of the valve actuator; the second ferromagnetic component and the first ferromagnetic component magnetically attract or repel each other; wherein, when the valve actuator When the moving end is in the open position, the first ferromagnetic component is driven by the second ferromagnetic component to open the bottom opening. A cell sub-processing machine used to collect the contents of a plurality of cell culture boxes into two centrifuge boxes. The cell sub-processing machine includes: a base; a box processing device located on the base, and It includes: a box positioning platform, which is rotatably or movably arranged on the base; a plurality of box receptacles, each of which is used to accommodate one of the cell culture boxes or one of the centrifuge boxes; each The box holder is rotatably provided on the box positioning platform, and can rotate to a tilting angle to pour out the contents of the cell culture box or the centrifuge box; a plurality of box driving assemblies that control the boxes respectively According to the angle of the box holder, each box driving assembly can swing the corresponding box holder or rotate the corresponding box holder to the tilting angle; wherein, the box positioning platform can move each box The body container seat moves to a first injection position; a liquid even distribution device is provided on one side of the box processing device and includes: a dispensing positioning platform that can rotate or move relative to the base; at least A gravity dispensing device as described in any one of claims 1 to 4, which is disposed on the dispensing positioning table; wherein the dispensing positioning table can move the at least one gravity dispensing device from the first dispensing position. The lower part of the medicine position is moved to the upper part of the two centrifuge boxes, so that the liquid in the funnel of the at least one gravity dispensing device can flow downward from the two output flow channels into the two centrifuge boxes respectively. The cell subculture processor as described in claim 5, wherein: The box positioning platform can move each box holder to a second injection position; the cell sub-processing machine has an injection device; the injection device is provided on the base and includes a plurality of injection heads ; Each injection head is used to connect a medicament, and can be moved above the second injection position to inject the connected medicament into the cell culture box located on the box holder at the second injection position. within. The cell subculture processor of claim 5, wherein the box positioning platform of the box processing device can move each box holder to a box receiving position or a box output position; The processor includes: a main box moving device, which is used to move the two centrifuge boxes located below at least one gravity dispensing device to the box receptacle located at the box receiving position; a centrifugal device, It is provided on one side of the box processing device; an auxiliary box moving device is used to move the centrifuge box in the box receptacle at the box output position into the centrifuge device, and is used to Move the centrifuge box in the centrifuge device to the box holder at the box output position. The cell subculture processor as described in claim 5, wherein: the box positioning platform of the box processing device can move each box holder to a box output position; the cell subculture processor includes: An optical detection device is provided on one side of the box processing device and is used to detect the number of cells in the cell culture boxes; an auxiliary box moving device is used to move the box at the output position of the box. The cell culture box in the box receptacle moves into the optical detection device, and is used to move the cell culture box in the optical detection device to the box receptacle at the box output position. The cell subculture processor as described in claim 7, wherein: The box positioning table of the box processing device is rotatably provided on the base, and can accommodate each box at the box receiving position, the box output position, the first injection position and cyclic movement between a second injection position; the cell subculture processor has an injection device, which is located on the base and includes a plurality of injection heads; each of the injection heads is used to connect a medicament, and can Move to above the second injection position to inject the connected medicament into the cell culture box located on the box holder of the second injection position; the cell subculture processor includes an optical detection device, It is provided on one side of the box processing device; the auxiliary box moving device is used to move the cell culture box in the box receptacle at the box output position to the optical detection device, and is used to Move the cell culture box in the optical detection device to the box receptacle at the box output position; wherein the second injection position and the box output position are respectively located opposite to the box processing device ; The centrifugal device, the auxiliary box moving device and the optical detection device are located on the side of the box processing device facing the box output position, and the injection device is located on the side of the box processing device facing the second injection position. one side. The cell subculture processor according to claim 5, wherein: the box positioning platform of the box processing device can move each box holder to a box receiving position; the cell subculture processor includes a The main box moving device is used to move the cell culture box located in the box receiving position of the box to below the at least one gravity dispensing device, so that the at least one gravity dispensing device The liquid in the funnel can flow into the cell culture box. A cell subculture processing method used to collect the contents of a plurality of cell culture boxes into two centrifuge boxes. The cell subculture processing method includes: using the gravity dispensing device described in any one of claims 1 to 4. The contents of the cell culture boxes are evenly distributed into the two centrifuge boxes, and then the two centrifuge boxes are placed at opposite positions in a centrifuge device and centrifuged.

Images