KR20210127344A - System for automatically incubating cell using a robot - Google Patents

Abstract

The present invention relates to a cell culture automation system using a robot. An automated cell culture system using a robot comprises: a moving unit for moving cell culture equipment including a culture vessel, a vessel for centrifugation, and a tube for a cell sample; a cell incubator for culturing cells in the culture vessel by receiving the culture vessel; a centrifuge for centrifuging the cultured cells; a cytometer for counting the number of centrifuged cells; a multi-joint robot; an air displacement pipette (ADP) which injects cells and a culture medium into the cell culture equipment or extracts cells and a culture medium from the cell culture equipment using air suction and exhaust; and a control unit which controls the multi-joint robot to pick up the culture vessel moved by the moving unit and place the same in the cell incubator, controls the multi-joint robot to pick up the culture vessel in the cell incubator when the cells in the culture vessel are cultured for a predetermined period, controls the ADP to put the contents of the culture vessel into the vessel for centrifugation, controls the multi-joint robot to deliver the centrifuge container to the centrifuge, controls the ADP to sample the centrifuged cells and put the same in the tube for a cell sample, controls the multi-joint robot to introduce the cells in the cell sample tube into the cell counter, and receives counted cell number information from a cytometer. Therefore, cells can be automatically mass-cultured.

Description

Cell culture automation system using a robot {System for automatically incubating cell using a robot}

The present invention relates to cell culture, and more particularly, to an automated cell culture system using a robot.

Cell culture is a very important technology in biological research including molecular biology, and refers to culturing specific cells for the purpose of diagnosing or treating human diseases. In recent years, it is required to efficiently mass-culture cells, tissues, microorganisms, viruses, etc. (collectively referred to as "cells") in the fields of pharmaceutical production, gene therapy, regenerative medicine, immunotherapy, and the like.

The immune system is regulated by a complex mechanism, and when there is an abnormality in the immune system, an imbalance in the immune system is caused and incurable diseases such as cancer occur. A tumor (cancer) is a tissue composed of autologous cells caused by mutations in normal cells by environmental and genetic factors, and progresses over a long period of time. The number is rapidly decreasing. The human body produces immune cells such as cytotoxic Tlymphocytes (CTL) or natural killer cells (NK cells) against these tumors to kill cancer cells.

For the treatment of cancer, an increase in the killing ability and activity of these immune cells in cancer patients is very important for the treatment of cancer. Chemotherapy is an important part of cancer treatment and aims to relieve symptoms by removing or reducing the size of the tumor. Here, there is a task to reduce recurrence or increase the survival rate, and above all, anticancer drug treatment should improve the quality of life of patients.

However, the current treatment does not yet approach a satisfactory cancer cure rate, and above all, it has weaknesses in resistance to anticancer drugs and side effects, and that the drugs used are not effective for everyone. In addition, improvement of the regimen, regulations, and administrative procedures related to the use of drugs for cancer treatment is necessary to the extent that the problem of improving the quality of life in cancer treatment in Korea is still the second problem. Therefore, there is a high demand for the development of a new treatment that is effective for each patient, has fewer side effects and can be received comfortably.

The conditions for the improved treatment should be systemic treatment and more effective, less toxicity and less effect on normal cells, and treatment focusing on the specific biological characteristics of cancer that distinguish it from normal cells. Accordingly, in recent years, clinical or therapeutic development using not only chemotherapy but also immunotherapy is being rapidly promoted.

Among them, NK cells in the immune system of the human body are one of the representative innate immune cells, and are known to have the ability to non-specifically kill cancer and to recognize and kill viruses and bacteria. The effect of chemotherapy using immune cells, such as NK cells, may vary depending on the progression of cancer. Immune cell therapy can maximize its effectiveness when used in patients at an early stage of cancer progression or to suppress cancer recurrence after resection surgery. Therefore, for the treatment of such cancer diseases, it is essential to maintain the cancer cell killing ability and activity of NK cells in cancer patients, and it is also important to maintain the number of NK cells.

Since the number of NK cells present in the body is not large, a technology for mass production while maintaining sufficient efficacy to use NK cells for therapeutic purposes is essential. However, NK cells are not properly proliferated and cultured in vitro (in-vitro). Therefore, attention is focused on technology for amplifying and culturing NK cells to a practically useful level, and although many studies are being conducted, it has not yet reached the level applicable to clinical practice.

For culturing of NK cells, a culturing technique is being developed that maximizes treatment by extracting NK cells from a patient, proliferating the extracted NK cells several times or tens of times, and then administering them to the patient again. It is difficult to mass-cultivate because it is performed manually under

Patent Publication No. 2019-0124608, 2019.11.05)

The problem to be solved by the present invention was created to solve the problems of the prior art. In a cell culture apparatus composed of a plurality of clean rooms (chambers) having an aseptic environment, using a robot to load cells into a cell incubator and operate the robot Automated cell culture using a robot, which can automatically cultivate cells in large quantities by maintaining the extraction and injection of cells at a certain amount and unloading the cells increased by culturing for a certain period using a robot to provide a system.

A cell culture automation system using a robot according to the present invention for achieving the above technical problem is a movement of moving cell culture equipment including at least a culture vessel containing cells to be cultured and a culture medium, a centrifugation vessel, and a cell sample tube wealth; a cell incubator for accommodating the culture vessel and culturing the cells in the received culture vessel; a centrifuge for centrifuging the contents of the centrifuge container; a cytometer for counting the number of cells by extracting a certain amount from the cell sample tube containing the contents of the centrifuged cells and the fresh culture solution; A multi-joint robot capable of moving up, down, left, right, back and forth, and rotatable; ADP (Air Displacement Pipette) for injecting cells and culture medium into the cell culture material or extracting cells and culture medium from the cell culture material using suction and discharge of air; and controlling the articulated robot to pick up the culture vessel moved by the moving unit and put it in the cell incubator, and when the cells contained in the culture vessel are cultured for a predetermined period, control the articulated robot to be in the cell incubator Pick up the culture vessel, control the ADP to put the contents of the picked up culture vessel into the centrifuge vessel, control the articulated robot to transfer the centrifuge vessel to the centrifuge, and the ADP Control to sample the cells centrifuged by the centrifuge and put them in the cell sample tube, control the articulated robot to put the cells in the cell sample tube into the cell counter, and count from the cell counter It includes a control unit for receiving the cell number information.

The automated cell culture system according to the present invention includes: a first chamber in which the control unit is located, the cell culture equipment is brought in and out, and an operating space of the cell culture system is provided; a second chamber in which the moving unit enters and exits and loads the vessel used for cell culture into the moving unit; and the cells are cultured by the cell incubator, the cultured cells and the solution are centrifuged by the centrifuge, the cultured cells are counted by the cell counter, and the cells cultured from the culture vessel by the ADP and a third chamber in which the culture medium is taken out, the cultured cells and the new culture medium are injected into another culture vessel, and the culture vessel, the centrifuge vessel, and the cell sample tube are moved by the multi-joint robot.

The cell culture automation system according to the present invention is equipped with a DECAP device that opens and closes the lids of the culture vessel, the centrifugation vessel and the cell sample tube moved by the multi-joint robot, and the ADP is mounted to move the ADP up, down, front and rear, left and right. A gantry robot that moves to a cell culture system chamber, a bacteria test container that collects bacteria in the cell culture system chamber, a sterilizer that sprays hydrogen peroxide water vapor into the cell culture system chamber, and the cells in the culture bag that have been cultured in the cell culture machine for centrifugation After centrifugation in a container and the centrifuged cells are transferred to the final bag, at least one of a tube sealer for sealing the final bag may be further included.

In the automated cell culture system according to the present invention, when the cells in the culture vessel are cultured in the cell incubator for a certain period of time, the cells in the culture vessel are accommodated by the multi-joint robot, the DECAP robot, and the ADP, a culture bag with a larger capacity than the culture vessel; and a mechanism for fixing the culture bag to facilitate movement of the culture bag by the articulated robot, and prevents distortion of the culture bag according to the movement of the culture bag and rotation of the culture bag when the lid of the culture bag is opened and closed It may be further provided with a culture bag jig.

The cell culture automation system according to the present invention is attached to the cell culture equipment, and at least an identification code issuer for issuing an identification code including information identifying the cultured cell donor, an identification code reader for reading the identification code, the It is mounted on a multi-joint robot, and photographs the type and quantity of cell culture equipment brought in through the light equipment table and the heavy equipment table, opens the lid of the culture bag, and opens the door of the cell incubator when opening the door of the cell incubator. It may further include a vision scan to photograph.

The moving unit includes a light-weight equipment table for accommodating light-weight equipment, including the culture vessel and the tube for the cell sample; and a weight equipment table for accommodating heavy equipment heavier than the light equipment, including the container for centrifugation, wherein the articulated robot is movable and rotatable up and down, left and right, and is moved by the first orthogonal robot A lightweight multi-joint robot that picks up and moves lightweight equipment; and a weight articulated robot capable of moving up and down, left and right and rotatable, and picking up and moving the heavy equipment moved by the second orthogonal robot.

According to the cell culture automation system and method using a robot according to the present invention, in a cell culture apparatus composed of a plurality of clean rooms (chambers) having a sterile environment, using a robot, the cells are loaded into the cell incubator, and the cells are loaded using the robot. It is possible to automatically cultivate large amounts of cells by maintaining the extraction and injection of the cells at a certain amount accurately, and by culturing the cells for a certain period and unloading the increased cells using a robot.

1 shows the appearance of an embodiment of an automated cell culture system using a robot according to the present invention.
2 is a block diagram showing the configuration of an embodiment of an automated cell culture system using a robot according to the present invention.
3 shows an example of a high-speed ADP.
4 shows an example of a culture bag and a culture bag jig.
Figure 5 shows that the culture bag is mounted on the culture bag jig.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings. The configuration shown in the embodiments and drawings described in this specification is only a preferred embodiment of the present invention, and does not represent all the technical spirit of the present invention, so various equivalents that can be substituted for them at the time of the present application It should be understood that there may be variations and variations.

1 shows the appearance of an embodiment of an automated cell culture system using a robot according to the present invention. Referring to FIG. 1 , the automated cell culture system using a robot according to the present invention is a first chamber 110 , a second chamber 120 , and a third chamber 130 that are a plurality of clean rooms having an aseptic environment. ) can be made. In particular, filters 142 , 144 , 146 , and 148 for removing particles having a predetermined size or larger such as dust in the air may be installed on the ceilings of the second chamber 120 and the third chamber 130 . As the filters 142 , 144 , 146 , and 148 , a High Efficiency Particulate Air Filter (HEPA) filter may be used. And in the first chamber 110 , the second chamber 120 , and the third chamber 130 , the air is supplied from top to bottom by equipment that purifies air such as an EFU (Equipment Fan Filter Unit) and supplies clean air. direction can be cycled. The first chamber 110 , the second chamber 120 , and the third chamber 130 may be managed with different GMP (Good Manufacturing Practice) grades. The second chamber 120 has a higher degree of cleanliness than the first chamber 110, so that the cleanroom class (grade) is different. An ultraviolet sterilizer may be installed on the ceiling of the second chamber 120 to remove bacteria that may be introduced from the first chamber 110 .

There is a trend that culture is carried out with an unmanned automated system within the configured culture equipment. That is, the extracted NK cells are loaded into the culture equipment and cultured under aseptic conditions until a predetermined date to unload the increased NK cells to the outside.

2 is a block diagram showing the configuration of an embodiment of an automated cell culture system using a robot according to the present invention. An embodiment of the cell culture automation system using the robot according to the present invention is a moving unit (210, 212), a cell culturer (220, 222), a centrifuge 230, a cell counter 240, a multi-joint robot (250). , 252 ), an Air Displacement Pipette (ADP) 260 , and a control unit 270 . In addition, an embodiment of the cell culture automation system using the robot according to the present invention is a DECAP robot 280 and a gantry robot 262, a bacteria test container 215, a sterilizer 225, a tube sealer 235, culture It may further include at least one of a bag and culture bag jig, an identification code reader (Reader, 245), an identification code issuer 255, and a vision scan (280, 282).

The moving parts 210 and 212 move the cell culture equipment including at least a culture vessel containing cells to be cultured and a culture solution, a centrifugation vessel, and a cell sample tube. The moving parts 210 and 212 may include a light equipment table 210 and a heavy equipment table 212 . The lightweight equipment table 210 accommodates lightweight equipment and may be moved by an orthogonal robot (not shown). The lightweight equipment includes the culture vessel (flask), syringe, and tube for the cell sample (small tube, micro tube), and the like.

The flask is used to contain the NK cells extracted from the blood, and when the flask containing the NK cells together with the cell medium is brought into the third chamber 130, it is placed in the cell incubator 250, 252 by the lightweight multi-joint robot 250. housed and cultivated. The NK cells in the flask are transferred to a culture bag after being cultured for a certain period of time in a cell incubator.

A small tube is a conical tube containing cells cultured in the cell incubator 220, 222, centrifuged through the centrifuge 230, and then a certain amount of the solution in the upper part of the conical tube is put into the solution. It can be used when taking out to check how many cells are included. In addition, the small tube may be used to contain the centrifuged cells from the conical tube for dilution of the centrifuged cell solution. The micro tube can be used to dilute a certain amount of cells in a small tube by mixing them with the cell medium. In addition, it may be used when containing a small amount of cell fluid in order to count the cells in the cytometer 240 .

Cell culture equipment brought in by the lightweight equipment table 210 is mainly handled by the lightweight articulated robot 250 . The heavy equipment table 212 accommodates heavy equipment heavier than the light equipment and may be moved by an orthogonal robot. Heavy equipment includes a container containing a cell medium, a container containing a washing liquid to wash out cells mixed with foreign substances, and a final solution, which is a liquid that is mixed at the end when cells that have been cultured are taken out. It includes a container in which it is contained, a conical tube used as the container for centrifugation, a waste medium container, a culture bag, and the like. Cell culture equipment brought in by the heavy equipment table 212 is mainly handled by the weight articulated robot 252 .

The cell incubator (Incubator, 220, 222) accommodates the cell culture vessel, and culture the cells contained in the received culture vessel. The cells may be, for example, NK cells, and the NK cells may be extracted from human blood. The culture vessel includes a flask and a culture bag. The flask accommodates the culture medium and NK cells extracted from human blood. And the culture bag accommodates a large amount of cultured NK cells when the NK cells contained in the flask are cultured for a certain period in the cell incubators 220 and 222 and the number and amount of cells increases.

The centrifuge 230 centrifuges the contents of the centrifuge container. The centrifugation vessel may be a conical tube.

A cell counter (Cell Counter, 240) counts the number of cells by withdrawing a certain amount from the cell sample tube containing the mixed contents of the centrifuged cells and the fresh culture medium. The cell counter counts the number of cells by taking a sample of the cells cultured in the cell incubator every predetermined period during the cell culture period. For example, after extracting the liquid in the upper part of the plurality of centrifuged conical tubes, the centrifuged cells remaining in the conical tubes are collected in one container and made to a certain amount by adding a medium to the ADP (260). mixed by A portion from a certain amount of mixed cells is placed in a tube for cell sample. A certain amount of cells in the cell sample tube is extracted and the number of cells is counted through the cell counter 240 . Cell counting can also be done through artificial intelligence. An image of the sampled cells is taken using a vision camera provided in the cytometer 240 . After learning based on a deep-learning-based algorithm considering the shape, size, color, living cell, dead cell, etc. of the cell from the captured image, the number of cells can be obtained through the sampled cell image. The number of counted cells is transmitted to the controller 270, and an amount of medium may be further added to manage the number of cells according to the culture period based on the counted number of cells.

The articulated robots 250 and 252 are movable and rotatable up, down, left and right, and include a lightweight articulated robot 250 and a heavy articulated robot 252 . The lightweight articulated robot 250 is movable and rotatable up and down, left and right, and picks up and moves the lightweight equipment on the lightweight equipment table 210 moved by the orthogonal robot. The multi-joint weight robot 252 is rotatable and moves up and down, left and right, and picks up and moves the heavy equipment on the heavy equipment table 212 moved by the orthogonal robot.

ADP (Air Displacement Pipette, 260) injects cells and culture medium into the cell culture material or extracts cells and culture medium from the cell culture material by using air suction and discharge. For example, the cells cultured in the cell incubators 220 and 22 are placed in a conical tube and centrifuged in the centrifuge 230 , and then the solution in the upper part of the conical tube is extracted using the ADP 260 . In addition, the ADP 260 may be used when mixing the centrifuged cells and medium into the culture vessel after extracting the liquid from the upper part of the centrifuged conical tube. In the embodiment of the present invention, various types of ADPs may be provided as the ADP 260 according to capacity and speed. For example, high-speed ADP with high volume and rapid inhalation and exhalation; low-speed ADP with high volume and low inhalation and exhalation; : 1ml) for ADP. 3 shows an example of the high-speed ADP 260 . 3, the high-speed ADP is made including the ADP body 310 and the tip (TIP, 320), the tip (TIP, 320) is a syringe shape with a scale indicating the amount of liquid to be sucked and discharged, It is detachable from the ADP body 310 . The high-speed ADP is capable of sucking and discharging a large amount of cell culture solution, and is capable of dividing a predetermined amount through multi-stage control and then discharging the desired amount step by step. and the culture medium can be uniformly mixed. Suction and discharge of the cell fluid using the ADP (260) may be made on the work table (265).

The control unit 270 controls the cell culture process in the automated cell culture system according to the present invention, and includes an input/output device such as a monitor and a keyboard. The control unit 270 controls the articulated robots 250 and 252, picks up the culture vessel moved by the moving unit 210, 212, puts it in the cell incubator 220, 222, and puts it in the culture vessel for a predetermined period. When the cells are cultured, the articulated robots 250 and 252 are controlled to pick up the culture vessels in the cell incubators 220 and 222 . The control unit 270 controls the ADP 260 to put the contents of the picked up culture vessel into the centrifuge vessel, and controls the articulated robots 250 and 252 to place the centrifuge vessel into the centrifuge 230 . The cells are transferred to, and the cells centrifuged by the centrifuge 230 are sampled by controlling the ADP 260 and placed in the tube for the cell sample. The controller 270 controls the articulated robots 250 and 252 to put the cells in the cell sample tube into the cytometer 240 and receives information on the number of counted cells from the cytometer 240 .

1 and 2 , the first chamber 110 is a space where an administrator manages and operates an automated cell culture system, and the control unit 270 is located therein. In the first chamber 110, an identification code capable of identifying the cell culture equipment is output through the identification code issuance machine 255, and the identification code (eg, QR code) is attached to the cell culture equipment and the container inlet ports 290 and 292. ), the cell culture equipment is brought into the second chamber 120 . In the second chamber 120, the moving parts 210 and 212, that is, the light equipment table 210 and the heavy equipment table 212 enter and exit, and the containers used for cell culture are loaded into the moving parts 210 and 212. .

In the third chamber 130, the cells are cultured by the cell incubator 220, 222, the cultured cells and the solution are centrifuged by the centrifuge 230, and the cultured cells are separated by the cell counter 240. counted, the cultured cells and the culture medium are taken out from the culture vessel by the ADP (260), the cultured cells and a new culture medium are injected into another culture vessel, and the culture vessel by the multi-joint robot (250, 252); A vessel for centrifugation and a tube for cell sample are moved.

The cell culture automation system using the robot according to the present invention further includes a DECAP robot 280 and a gantry robot 262, a bacteria test container 215, a sterilizer 225, a tube sealer 235 and a scale 285. can do. The DECAP robot 280 opens and closes the lids of the culture vessel, the centrifuge vessel, and the cell sample tube moved by the articulated robots 250 and 252 .

The gantry robot 262 is equipped with an ADP 260 to move the ADP 260 , for example, a high-speed ADP, a low-speed ADP, a small amount ADP, and a very small amount ADP up, down, front, back, and left and right directions. The bacteria test container 215 collects bacteria in the chamber of the cell culture system, and may be replaced in units of a predetermined period. The bacteria test container that has been replaced and taken out is cultured and checked for bacteria.

The sterilizer 225 sprays hydrogen peroxide water vapor into the second chamber 120 and the third chamber 130 of the cell culture system to remove bacteria in the chamber. The tube sealer (235) centrifuges the cells in the culture bag that have been cultured in the cell incubator 220 in a centrifugation container, and when the centrifuged cells are transferred to the final bag, the final bag is sealed. (sealing) The scale 285 is used to measure whether the solution required for cell culture is properly transferred from one container to another. By using the scale 285, it is possible to recognize the exact amount of the moved solution when moving the solution between different containers, and also check whether an omission has occurred.

The cell culture automation system using the robot according to the present invention may further include a culture bag and a culture bag jig, an identification code issuer 255 , an identification code reader 245 , and vision scans 280 and 282 .

4 shows an example of a culture bag and a culture bag jig. Figure 5 shows that the culture bag is mounted on the culture bag jig. 4 and 5, the culture bag 420 has a larger capacity than the culture vessel and is flexible, and when the cells in the culture vessel are cultured for a certain period in the cell incubator 220, the articulated robot Cells in the culture vessel are accommodated by (250, 252), the DECAP robot 280 and the ADP 260, and a plastic bag can be used as a culture bag.

The culture bag jig 420 supports the flexible culture bag stably, and the movement of the culture bag by the articulated robots 250 and 252 and the cultured cells by the ADP 260 and the DECAP robot 280 are performed. It is a device that facilitates injection and extraction. That is, by stably supporting the upper and lower portions of the culture bag through the culture bag jig, the articulated robot (250, 252) dp prevents distortion of the culture bag according to the movement of the culture bag, and ADP (260) and DECAP robot (280) Prevents the culture bag from rotating together with the lid when the lid (CAP) of the culture bag is opened and closed by

The identification code issuer 255 is located in the first chamber 110, and when the cell culture equipment is brought into the second chamber 120 through the light equipment table 210 and the heavy equipment table 212, the cell culture Identification codes attached to equipment, that is, light equipment and heavy equipment, are issued. The identification code includes at least information identifying the cell donor to be cultured, and information such as gender, age, disease, and culture period of the cell donor may be included, and the identification code information may be input through the control unit 270 . can Examples of the identification code may include a barcode or a QR code.

The identification code reader (BCR, 245) reads the identification code attached to the cell culture equipment. The identification code reader 245 reads the identification code attached to the cell culture equipment when identifying the cells contained in the cell culture equipment in the cell culture process when the cells of a plurality of donors are cultured together in the cell culture system. Control unit 270 send to

The vision scan (280, 282) is to photograph the type and quantity of cell culture equipment brought in through the light equipment table 210 and the heavy equipment table 212, open the lid of the culture bag, and inject a syringe ( Tip) is mounted, and when the door of the cell incubator 220, 222 is opened, the door of the cell incubator 220, 222 is photographed to check whether the door of the cell incubator is properly opened, and the photographed image is controlled by the control unit ( 270). The vision scans 280 and 282 may be mounted on the lightweight articulated robot 250 and the heavy articulated robot 252 .

An operation of the automated cell culture system using a robot according to the present invention will be described with reference to FIGS. 1 and 2 . In the automated cell culture system using a robot according to the present invention, cells to be cultured, for example, NK cells extracted from blood, are loaded into the culture system and cultured in the cell incubator 220 and 222 under aseptic conditions until a predetermined date. This is to unload the NK cells with an increased number of cells to the outside. That is, the NK cells contained in the flask are cultured for a set period in the cell incubator 220, 222, and when the number of cells in the flask increases, the NK cells are centrifuged to discard the unnecessary solution, and the precipitated NK cells are placed in a culture bag with the medium and They are injected together and put into the cell incubators 220 and 222 for re-cultivation. The above culture process is performed periodically until a set period is reached, and cultured NK cells are obtained through this series of processes.

More specifically, when describing the culturing process of NK cells, the extracted NK cells are placed in a flask together with the medium, and light and heavy equipment necessary for cell culture through the light container inlet 290 and the heavy container inlet 292 are used. together with the moving parts 210 and 212 to the second chamber 120 and the third chamber 130 . At this time, the identification code issued through the identification code issuing machine 255 is attached to light equipment and heavy equipment including the flask.

When a flask containing NK cells is brought in, the articulated robot (250, 252) accommodates the flask in the cell incubator (220, 222) to culture the NK cells for a certain period of time. At this time, the cell culture materials in the light equipment table 210 and the heavy equipment table 212 are unloaded by the articulated robots 250 and 252 and are vision-scanned by the vision scanners 280 and 282. The presence and quantity of cell culture equipment is checked. Then, the identification code attached to the cell culture equipment is read by the identification code reader 245, and the doors of the cell culture devices 220 and 222 are vision-scanned, and then the inner and outer doors of the cell culture devices 220 and 222 are opened. , flasks containing NK cells are accommodated in cell incubators 220 and 222 by articulated robots 250 and 252, and NK cells are cultured.

When the NK cells are cultured for a certain period of time, the articulated robots 250 and 252 pick up the flasks from the cell incubators 220 and 222 and move them to the work table 265 . When the identification code attached to the flask is confirmed by the identification code reader 245, the lid (CAP) of the flask is opened by the DECAP robot 280, and the NK cell-free solution is discharged into the flask by the ADP 260, and Add new culture medium. The flask into which the new culture solution is introduced is put into the cell incubators 220 and 222 by the articulated robots 250 and 252, and cultured again.

After a certain period of time, when the cell culture equipment including the culture bag is brought in, the quantity is checked through the vision scanner (280, 282), and the single-joint robot (250, 2452) is equipped with NK cells cultured from the cell incubator (220, 222). When the flask is picked up, the contents of the flask are moved to a conical tube, a centrifugation vessel, by the ADP (260) and the DECAP robot (280). The conical tube in which the cultured NK cells are transferred is centrifuged when seated in the centrifuge 230 by the articulated robots 250 and 252. At this time, a balance vessel may be added to the centrifuge 230 for centrifugation. After centrifugation, the culture solution at the top of the conical tube is removed by the articulated robots 250 and 252, the ADP 260 and the DECAP robot 280, and diluted with a solution for cell count to count the cells. At 240 the number of viable cells is counted. The cell count may be repeated several times. The NK cells in the conical tube are collected in one container, and when the culture medium is added, they are mixed and transferred to the culture bag. In the middle of this process, the bacteria in the third chamber 130 can be managed through the bacteria test container 215 .

The process of cell culture in the cell incubators 220 and 222, centrifugation of the cultured cells in the centrifuge 230, and the cell counting process of the centrifuged cells in the cytometer 240 may be repeated several times.

When the cell culture is completed, it is put into the final bag, sealed through the tube sealer 235, and then taken out for distribution.

The control unit 270 controls each process of cell culture, and inputs equipment and culture medium required for cell culture by using information on the number of cultured cells. Then, if there is a problem, the input solution is judged according to the number and concentration of the cells, and the culture medium is determined by tracking the growth trend of the cells, identifying the cell type, the number of cells, the number of living cells, the number of dead cells, and debris. For cell counting, sample collection is not done directly from the culture bag, but transferred to a small container, centrifugation can be performed every culture cycle, and the culture bag can be replaced.

Although the present invention has been described with reference to the embodiment shown in the drawings, which is merely exemplary, those skilled in the art will understand that various modifications and equivalent other embodiments are possible therefrom. Accordingly, the true technical protection scope of the present invention should be determined by the technical spirit of the appended claims.

110: first chamber 120: second chamber
130: third chamber 142, 144, 146, 148: filter
210: light equipment table 212: heavy equipment table
215: bacteria test vessel 220, 222: cell incubator
225: sterilizer 230: centrifuge
235: tube sealer 240: cell counter
245: identification code reader 250: lightweight multi-joint robot
252: heavy articulated robot 260: ADP
262: gantry robot 265: working table (working table)
270: control unit 280, 282: vision scan
285: scale 290: light container inlet
292: weight container inlet 310: ADP body
320: tip (TIP) 410: culture bag jig
420: culture bag

Claims (10)

a moving unit for moving cell culture equipment including at least a culture vessel containing cells to be cultured and a culture medium, a vessel for centrifugation, and a tube for a cell sample;
a cell incubator for accommodating the culture vessel and culturing the cells in the received culture vessel;
a centrifuge for centrifuging the contents of the centrifuge container;
a cytometer for counting the number of cells by extracting a certain amount from the cell sample tube containing the centrifuged cells and the mixed contents of the fresh culture solution;
A multi-joint robot capable of moving up, down, left, right, back and forth, and rotatable;
ADP (Air Displacement Pipette) for injecting cells and culture medium into the cell culture material or extracting cells and culture medium from the cell culture material using suction and discharge of air; and
Control the articulated robot to pick up the culture vessel moved by the moving unit, put it in the cell incubator, and when the cells contained in the culture vessel are cultured for a predetermined period of time, control the articulated robot to culture in the cell incubator take courage,
The contents of the culture vessel picked up by controlling the ADP are put into the centrifugation vessel, the multi-joint robot is controlled to deliver the centrifuge vessel to the centrifuge, and the ADP is controlled to the centrifuge. to sample the centrifuged cells and put them in the tube for the cell sample,
and a control unit for controlling the articulated robot to put the cells in the cell sample tube into the cell counter, and receiving information on the number of cells counted from the cell counter. According to claim 1, wherein the cell culture automation system
a first chamber in which the control unit is located, carrying in and out of the cell culture equipment, and providing an operating space of the cell culture system;
a second chamber in which the moving unit enters and exits and loads the vessel used for cell culture into the moving unit; and
The cells are cultured by the cell incubator, the cultured cells and the solution are centrifuged by the centrifuge, the cultured cells are counted by the cell counter, and the cells cultured from the culture vessel by the ADP and The culture medium is taken out, the cultured cells and the new culture medium are injected into another culture vessel, and the culture vessel, the centrifuge vessel, and the cell sample tube are moved by the multi-joint robot, characterized in that it consists of a third chamber Cell culture automation system. According to claim 1,
Cell culture automation system, characterized in that it further comprises a DECAP device for opening and closing the lids of the culture vessel, the centrifugation vessel and the cell sample tube moved by the articulated robot. According to claim 1, wherein the moving unit
a light-weight equipment table for accommodating light-weight equipment, including the culture vessel (flask) and the cell sample tube (small tube, micro-tube); and
Further comprising a weight equipment table for accommodating weight equipment heavier than the light equipment, including the container for centrifugation (conical tube),
The articulated robot
A lightweight multi-joint robot capable of moving up and down, left and right, and rotatable, and picking up and moving light equipment moved by the first orthogonal robot; and
A cell culture automation system comprising a weight articulated robot capable of moving up and down, left and right and rotatable, and picking up and moving the heavy equipment moved by the second orthogonal robot. According to claim 1,
The cell culture automation system, characterized in that it further comprises a gantry robot to which the ADP is mounted to move the ADP up, down, front and rear, left and right. According to claim 1,
a bacteria test container for collecting bacteria in the cell culture system chamber; and
Cell culture automation system, characterized in that it further comprises a sterilizer for spraying hydrogen peroxide water vapor into the cell culture system chamber. According to claim 1,
When the cells in the culture vessel are cultured in the cell incubator for a certain period of time, the cells in the culture vessel are accommodated by the articulated robot, the DECAP robot, and the ADP, and a culture bag having a larger capacity than the culture vessel; and
It consists of a mechanism that fixes the culture bag to facilitate movement of the culture bag by the articulated robot, and prevents distortion of the culture bag according to the movement of the culture bag and rotation of the culture bag when the lid of the culture bag is opened and closed Cell culture automation system, characterized in that it further comprises a culture bag jig. According to claim 1,
The cells in the culture bag, which have been cultured in the cell incubator, are placed in a centrifuge container and centrifuged, and when the centrifuged cells are transferred to the final bag, a tube sealer that seals the final bag is applied. Cell culture automation system, characterized in that it further comprises. According to claim 1,
an identification code issuer attached to the cell culture equipment and issuing an identification code comprising at least information identifying the cultured cell donor; and
Further comprising an identification code reader for reading the identification code, cell culture automation system. According to claim 1,
It is mounted on the multi-joint robot, and the type and quantity of cell culture equipment brought in through the light equipment table and the heavy equipment table are photographed, the lid of the culture bag is opened, and the door of the cell incubator is opened when the door is opened. A cell culture automation system further comprising a vision scan to take a picture.

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