JP2007295827A - Automatic culturing device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To improve the traceability of a culturing equipment used in an automatic culturing device. <P>SOLUTION: This automatic culturing device is constituted by being equipped with a multiple number of culturing equipment 21 to 24 and 40 including a culturing tool 1 used for cell culture, a reading means 46 for reading a discrimination label attached to at least one of the culturing equipment and a memory means for memorizing the discriminating label read by the reading means as associated with the information related to the cultured cells, and therefore, in the case that there occurs any inconvenience in the cells after the culture, it becomes possible to investigate whether the presence of the inconvenience is in the culturing equipment such as the culturing tool, etc., or not. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to an automatic culture apparatus, and more specifically to an automatic culture apparatus that can improve the traceability of a culture instrument.

  In recent years, research such as regenerative medicine has been actively conducted, and stem cell culture for reconstructing a living tissue has been performed. In this case, since the cultured cells are transplanted to the subject, safety such as contamination (contamination), cytotoxicity of chemical substances eluted from incubators such as petri dishes used for culturing operations, and other culture devices, etc. Careful consideration is necessary.

  For example, cell culture work is performed by exchanging the medium in the incubator or subculture to optimize the cell density in order to obtain many cultured cells, and these cell culture work avoids the occurrence of contamination. Therefore, it is usually performed carefully in a relatively clean atmosphere in which airborne particulates are reduced. For example, in manual culture, lift the lid of the incubator, such as a petri dish, during work such as medium exchange or subculture, and take care not to mix bacteria, and remove the sterile pipette between the petri dish and its lid. In addition, the pipette works quickly so that it does not touch surrounding objects such as the edge of the petri dish.

  However, in manual culture, it is not sufficient to completely avoid contamination even in a clean atmosphere, so do not open / close the incubator lid or use a pipette in an environment where the cells do not touch the atmosphere. An automatic culture apparatus capable of performing a culture operation has been proposed (for example, Patent Documents 1 and 2).

  On the other hand, in cell culture, since culture devices that directly touch the drug, such as a culture vessel such as a petri dish, a container for the drug such as a medium, or a tube through which the drug passes, those culture tools do not adversely affect the cell culture. So care is taken. For example, these culture instruments are made of a non-cytotoxic material such as polystyrene or polyethylene terephthalate, and are used after being sterilized. However, even if the material has the same name, there are actually various grades depending on the subtle composition of the raw material composition. Therefore, a cytotoxicity test and a sensitization test are usually performed for each lot of the raw material. As sterilization methods, sterilization using EO (ethylene oxide) gas or γ-ray is known and widely used.

JP 2004-016194 A JP 2004-229619 A

  By the way, the test items and test contents of the cytotoxicity test and sensitization test of the culture apparatus, and the sterilization method are important matters that directly affect the safety of the cultured cells, and therefore have extremely high traceability. Required. That is, it is required that the history of the culture instrument used for the cell culture can be confirmed by the recorded material.

  Usually, for culture devices that come into contact with drugs such as cultured cells and culture media, those that have been properly tested and sterilized by the manufacturer or seller are provided to users who perform cell culture, so traceability is ensured. ing.

  However, there is a problem that traceability is not always considered at the stage of actual cell culture. For example, it is possible to mistakenly use an improper culture instrument that has not been fully tested and sterilized in an automated culture device. In this case, the cultured cell may be contaminated with a cytotoxic substance or other bacteria, and the safety of the cultured cell may be impaired. In addition, culture devices that come into contact with cultured cells or drugs should be replaced with new ones at the end of each culture in order to prevent contamination by other patients' cells. And there is a risk of damaging the safety of the cultured cells.

  In these cases, since the conventional automatic culture apparatus cannot cope, there is a problem that when the safety of the cultured cells is impaired, the cause cannot be investigated retrospectively.

  This invention makes it a subject to improve the traceability of the culture instrument used with an automatic culture apparatus.

  In order to solve the above-described problems, an automatic culture apparatus of the present invention includes a plurality of culture instruments including an incubator used for cell culture, a reading unit that reads an identification mark attached to at least one of the culture instruments, The identification mark read by the reading means comprises storage means for storing in association with information relating to cultured cells.

  First, a culture instrument that comes into contact with cultured cells or drugs, such as incubators, drug containers (bags), tubes, etc., is attached with a letter, symbol, barcode, or a combination identification mark by stamping or pasting the identification mark. Keep a record of the material of the culture device, the appropriateness test, the sterilization treatment, etc. Then, when carrying out cell culture, the identification mark attached to the culture instrument is read and stored in association with information on the cultured cells, for example, patient name, culture history, and the like. As a result, when any defect occurs in the cultured cells, it is possible to investigate whether or not the defect exists in a culture instrument such as a culture vessel, and the traceability of the culture instrument can be improved.

  In this case, provided is a determination means for determining whether or not the identification mark read by the reading means matches a registered identification mark stored in advance, and if the identification mark does not match the registered identification mark, It is preferable to warn to that effect. According to this, it is possible to prevent the cell culture from being erroneously performed using an inappropriate culture instrument that has not been sufficiently tested and sterilized. In addition, when the cell culture is performed as it is, ignoring the warning, and any defect occurs in the cultured cells, it can be estimated with high accuracy that the location of the defect is in the culture instrument.

  In addition, a determination unit for determining whether or not the same identification mark as the identification mark newly read by the reading means is already stored in the storage means is provided, and the same identification mark as the newly read identification mark is stored. If it is already stored in the means, it can be configured to warn to that effect. According to this, it is possible to avoid a possibility that a culture instrument used for cell culture of a certain patient is erroneously reused for cell culture of another patient. Further, if the cell culture is carried out as it is, ignoring the warning, and some trouble occurs in the cultured cells, it can be estimated with high accuracy that the location of the problem lies in the culture instrument.

  Furthermore, the automatic culture apparatus of the present invention includes a plurality of culture devices including an incubator used for cell culture, control means for controlling the cell culture by operating the culture devices according to a culture procedure, and at least one of the culture devices. Reading means for reading the identification mark attached to the reading means, and determination means for determining whether or not the identification mark read by the reading means matches a registered identification mark stored in advance. Is preferably configured to output a cell culture start permission command to the control means only when the identification mark matches the registered identification mark. According to this, when an inappropriate culture instrument is used, it is possible to prevent any problems from occurring in the cultured cells by disabling the execution of the culture process. In addition, since automatic culture can be performed using only a culture instrument such as a culture vessel in which safety is ensured, the safety of a subject to whom the cultured cells are transplanted can be ensured.

  ADVANTAGE OF THE INVENTION According to this invention, the traceability of the culture instrument used with an automatic culture apparatus can be improved.

  Hereinafter, embodiments of an automatic culture apparatus to which the present invention is applied will be described.

(Embodiment 1)
An embodiment of the present invention will be described with reference to FIGS. 1 is a detailed view of an incubator according to an embodiment to which the present invention is applied. FIG. 1 (A) is a perspective view, FIG. 1 (B) is a side view, and FIG. 1 (C) is (B). It is sectional drawing in CC. FIG. 2 is an overall configuration diagram of an automatic culture apparatus according to an embodiment of the present invention. FIG. 3 is a block diagram of the present embodiment centering on the control device. 4A and 4B are flowcharts of an example of the automatic culture operation, and FIG. 5 is a flowchart of the processing procedure of the characteristic part. FIG. 6 is an example of recording information on cultured cells.

  As shown in FIG. 1, the incubator 1 is formed in a sealed container shape, and includes an injection nozzle 3 for injecting cells or drugs into the side wall, a discharge nozzle 4 for discharging cell liquid or waste liquid, and air suction. An exhaust nozzle 5 is provided. The injection nozzle 3 and the discharge nozzle 4 are provided with their tips positioned at the bottom in the incubator 1, and the intake / exhaust nozzle 5 is provided in the gas phase in the incubator 1. The material of the incubator 1 is not particularly limited, but is required to have cell non-toxic characteristics. For example, polystyrene and polyethylene terephthalate are preferable. The shape of the incubator 1 is not limited to this embodiment. In particular, a unique identification mark 6 is attached to the lid of the incubator 1. In the present embodiment, characters “ABCD corp.” Are engraved.

  The incubator 1 configured as described above is installed and used in an incubator 20 as shown in FIG. In the incubator 20, the temperature and the carbon dioxide concentration are adjusted to predetermined values. The incubator 1 is supplied with medicines from bags 22, 23, and 24 containing medicines through tubes 21 connected to the injection nozzle 3 of FIG. In the case of this embodiment, the bag 22 contains a cleaning solution, the bag 23 contains a release agent that decomposes cell surface adhesion proteins, and the bag 24 contains a culture medium. Pinch valves 25, 26, and 27 are provided on the branch tubes that connect the bags 22, 23, and 24 and the tube 21, respectively. The tube 21 is provided with a heater 28 for preheating a medicine such as a medium to be injected into the incubator 1, a pinch valve 29, and a pump 30. A liquid reservoir 31 is connected to the tube 21 between the pinch valve 29 and the pump 30 via a branch tube. The upper opening of the liquid reservoir 31 is covered with a film 32. Cells are injected into the liquid reservoir 31 through the film 32 from the syringe 33. A pinch valve 34 is provided in the branch tube to which the liquid reservoir 31 is communicated. In the incubator 20, air filters 35, 36 and 37 opened in the incubator 20 are provided. The air filter 35 is communicated with the tube 21. The air filters 36 and 37 are communicated with the intake and exhaust nozzle 5 of the incubator 1 through tubes provided with pinch valves 38 and 39, respectively. The air filters 35 and 37 prevent outside air in the incubator 20 from directly entering the inside of the incubator 1, and the air filter 36 prevents cells and the like inside the incubator 1 from entering the incubator 20. It is something to prevent.

  Further, the incubator 1 is connected to a bag 41 into which the waste liquid and a bag 42 into which the cultured cell liquid is to be placed through a tube 40 connected to the discharge nozzle 4 in FIG. The tube 40 is provided with a pump 43. The tube 40 communicates with the bags 41 and 42 via branch tubes on the downstream side of the pump, and pinch valves 45 and 46 are provided with the branch tubes. Further, an image acquisition means 46 such as a microscope with a CCD camera for observing the state of cells inside the incubator 1 is provided. Although not shown in FIG. 2, the pinch valve, the pump, and the like are sequence-controlled by the control device in accordance with the set procedure of the cell culture operation.

  Here, since the incubator 1, the bags 22 to 24, 41, 42 and the tubes 21, 40 and the branch tube are culture devices that come into contact with cells or drugs, they must be non-toxic to cells, for example, vinyl chloride, silicon, etc. The product is preferred. The pumps 30 and 43 are preferably of a type in which outside air does not enter, and a roller pump that squeezes a tube and feeds liquid is preferable.

  With reference to FIG. 3, the structure of the characteristic part of this invention is demonstrated. In the figure, the pinch valves 25 to 27, 29, 34, 38, 39, 44, 45 and the pumps 30, 43 of the automatic culture apparatus shown in FIG. This is indicated by a pump group 62. As shown in the figure, the heater 28, the pinch valve group 61, the pump group 62, and the image acquisition means 46 are controlled by a control unit 63 configured by a computer or the like. Further, the acquired image data is input to the control unit 63 from the image acquisition means 46.

  Further, the barcode reading means 64 is connected to the control unit 63 so that the read barcode is inputted. The reference image data storage unit 65 is connected to the control unit 63 via the comparison / determination unit 66. The reference image data storage unit 65 includes a registration identification mark indicating that the incubator 1, bags 22 to 24, 41 and 42, tubes 21 and 40, and branch tubes used in this embodiment are appropriate culture instruments. Images are stored. The control unit 63 is connected to a database unit 67, a counter unit 68, an input unit 69 such as a keyboard, and a display unit 70 such as a display.

  The control unit 63 configured as described above controls the heater 28, the pinch valve group 61, the pump group 62, the image acquisition unit 46, the barcode reading unit 64, the comparison / determination unit 66, and the like according to the culture process. It is supposed to be. In addition, patient information, cell types, and detailed conditions of the culture process are input using the input unit 69. In addition, the progress of the culture is displayed on the display unit 70.

  The operation of the automatic culture apparatus of the present embodiment will be described along the flow chart of the culture operation shown in FIGS. 4A, 4B and 5.

  First, after assembling culture apparatuses such as the incubator 1, bags 22 to 24, 41 and 42, the liquid reservoir 31, the filters 35, 36 and 37, the tubes 21 and 40, and the branch tube according to the configuration of FIG. 3, an automatic culture operation start command is input from the input unit 69. Thereby, the control part 63 controls operation | movement of an automatic culture apparatus according to the procedure shown in FIG. 4 based on the program of the automatic culture operation stored in the database part.

{Step 0}: Confirmation of Culture Instrument When a start command is input, the control unit 63 confirms whether or not the culture instrument to be used is appropriate according to the procedure shown in FIG. Here, although it demonstrates as what confirms whether the culture device 1 is an appropriate thing, it can confirm similarly about another culture instrument. In other words, since culture devices such as bags, tubes, and incubators are disposable, it is confirmed whether or not the culture devices are appropriate before setting them in the apparatus and starting culture.

  First, the control unit 63 sends a command to the image acquisition means 46 to image the incubator 1 and captures the read image (step S21). As shown in the figure, the incubator 1 is stamped with an identification mark 6 of the letters “ABCD corp.”. Next, the control unit 63 sends the read image to the comparison / determination unit 66, reads the reference image of the registered identification mark from the reference image data storage unit 65, compares the read image with the reference image, and determines the identification mark of the read image. It is determined whether there is a match with the registered identification mark (step S22). Next, when the identification mark of the read image matches the registered identification mark, the start of the culturing operation is permitted, and as shown in FIG. 6, the identification mark 6 is associated with the information about the cultured cells in the database unit 67. (Step S23, 24). Information such as cell origin information in FIG. 6 is input from the input unit 69 to the database unit 67 before culturing.

  On the other hand, if the identification mark of the read image does not match the registered identification mark, a warning that the incubator 1 has a problem is output to the display unit 70, and the process is terminated without starting the culture operation (step) S25).

  As described above, according to the present embodiment, when an inappropriate incubator 1 is used, it is possible to disable the culture process of the automatic culture apparatus, and safety is improved.

  In addition, after the cultured cells are transplanted into the patient, it is possible to follow up whether or not the cause is in the incubator 1 or the like in the case where some trouble occurs in the cells. Should there be a problem with the material such as the incubator 1, it is possible to immediately advise all users to cancel the use and prevent the spread of trouble.

{Step 1}: Medium injection When the start of the culture operation is permitted in Step 0, the control unit 63 opens the pinch valves 27, 29, and 38, operates the pump 30, and transfers from the bag 24 to the incubator 1. Inject medium. At this time, the air inside the incubator 1 is exhausted from the filter 36 in the direction of arrow B through the pinch valve 38, so that the medium can be injected without being clogged.

{Step 2}: Cell liquid injection After the medium injection is completed, the process proceeds to Step 2 where the syringe 33 filled with the cell liquid in which the cells to be cultured are suspended in the medium or the like is placed in the liquid reservoir 31. The film 32 is punctured to inject cells. Thereafter, the pinch valves 34 and 38 are opened, the pump 30 is operated, and the cell liquid in the liquid reservoir 31 is sent to the incubator 1. At this time, the air inside the incubator 1 is exhausted from the filter 36 in the direction of arrow B, so that the cell fluid can be injected without being clogged.

{Step 3}: Culture After injecting the cells, the process proceeds to Step 3 where the incubator 1 in the incubator 20 is allowed to stand and cultured for a predetermined period (eg, about 3 to 7 days).

{Step 4}: Air feed Since the incubator 1 is in a hermetically sealed state, it is necessary to ventilate the inside of the incubator 1 in a timely manner. Therefore, in the step, the pinch valves 29 and 38 are opened, the pump 30 is operated, and the filter 35 The air in the incubator 20 is sucked through the and the air is injected into the incubator 1. At this time, the air in the incubator 1 is exhausted in the direction of arrow B through the filter 36. In this figure, the air feed of step 4 is performed once, but it may be performed any number of times other than the medium exchange and cell recovery described later, and it is always performed continuously. Also good.

{Step 5}: Medium Replacement Step 5 is a step performed periodically (for example, every 3 to 7 days) for supplementation of cell metabolites eluted in the medium and nutrients in the medium. The period of medium exchange can be set in advance or can be arbitrarily set according to the state of cell proliferation. The number of medium exchanges is not particularly limited, but is set in advance in the present embodiment. The medium exchange includes a medium discharge for discharging the medium in the incubator 1 and a medium injection for injecting a new medium into the incubator 1.

  To discharge the medium, the pinch valves 39 and 44 are opened, the pump 43 is operated, and the medium inside the incubator 1 is discharged to the bag 41. At this time, since outside air is injected into the incubator 1 from the filter 37, the medium can be discharged from the incubator 1 without clogging.

  In the medium injection, the pinch valves 27, 29, and 38 are opened, the pump 30 is operated, and the medium is injected from the bag 24 into the incubator 1. At this time, the air inside the incubator 1 is exhausted from the filter 36 in the direction of arrow B through the pinch valve 38, so that the medium can be injected without being clogged.

{Step 6}: Confirmation of the number of medium exchanges Here, the number of medium exchanges is confirmed, and if the preset number of medium exchanges has been reached, the process returns to the next step 7, and if not, the process returns to step 4.

{Step 7}: Air feed The operation is the same as that in Step 4, the pinch valves 29 and 38 are opened, the pump 30 is operated, and air is injected into the incubator 1. The exhaust from the incubator 1 is exhausted in the direction of arrow B through the filter 36.

{Step 8}: Culture The same as step 3, and the culture is allowed to stand in an incubator for a predetermined period (for example, about 3 to 7 days).

{Step 9}: Collection of Cultured Cells Pinch valves 29, 38, 45 are opened and pump 30 is operated to inject the cleaning solution for bag 22 into incubator 1. Since the air inside the incubator 1 is exhausted in the direction of arrow B by the filter 36, the cleaning liquid can be injected without clogging. Thereafter, the pinch valves 39 and 44 are opened, the pump 43 is operated, and the cleaning liquid inside the incubator 1 is discharged to the bag 41.

  Next, the pinch valves 26, 29, and 38 are opened, and the pump 30 is operated to inject the release agent from the bag 23 into the incubator 1. Thereafter, the pinch valve 27 is opened, the pump 30 is operated, and the culture medium in the bag 24 is injected into the incubator 1. At this time, the air inside the incubator 1 is exhausted in the direction of arrow B by the filter 36, so that the medium can be injected without clogging.

  Thereafter, the pinch valve 45 is opened, the pump 43 is operated, and the cells inside the incubator 1 are injected into the bag 42. After the injection, the connected tube is cut with a heat seal to complete a series of cell cultures, and the cultured cells collected in the bag 42 are used.

  As described above, according to the present embodiment, when an inappropriate incubator 1 is used, the automatic culture apparatus can disable the execution of the culture process, and safety is improved. In addition, the present invention is not limited to the incubator, but is attached to a culture instrument that comes into contact with cultured cells or drugs, such as a bag or tube, with a letter, a symbol, a barcode, or an identification mark that is a combination thereof by stamping or pasting, Corresponding to the identification mark, a record of the material of the culture device, proper test, sterilization treatment, etc. is kept. Then, when carrying out cell culture, the identification mark attached to the culture instrument is read and stored in association with information on the cultured cells, for example, patient name, culture history, and the like. As a result, when any defect occurs in the cultured cells, it is possible to investigate whether or not the defect exists in a culture instrument such as a culture vessel, and the traceability of the culture instrument can be improved.

  For example, after a cultured cell is transplanted into a patient, if any defect occurs in the cell, it is possible to follow up whether the cause is in the incubator 1 or the like. Should there be a problem with the material such as the incubator 1, it is possible to immediately advise all users to cancel the use and prevent the spread of trouble.

  In addition, if a warning is issued when an inappropriate incubator 1 is used, the operator can recognize an error and replace it with an appropriate incubator 1, thereby avoiding a culture failure. be able to. In addition, since automatic culture can be performed using only a culture instrument such as a culture vessel in which safety is ensured, the safety of a subject to whom the cultured cells are transplanted can be ensured. If the cell culture is performed as it is, ignoring the warning, and the cell after culture has some trouble, it can be estimated with high accuracy that the location of the trouble is in the culture instrument.

  Further, the same effect can be obtained by pasting a barcode on a culture instrument such as the incubator 1 and reading the barcode with the barcode reading means 64.

(Embodiment 2)
FIG. 7 shows a flowchart of the processing procedure of the characteristic part according to another embodiment of the present invention. The present embodiment is different from the flowchart of FIG. 5 in that, in the determination of step S23, when the identification mark of the read image matches the registered identification mark, this reading is performed before permitting the start of the culture operation. Step S32 is provided for determining whether or not there is an identification mark that matches the identification mark of a culture instrument used in the past. Since other functions and configurations are the same as those of the first embodiment, the same reference numerals are given and descriptions thereof are omitted.

  In the determination in step S31, the past culture record in the database unit 67 is searched, and if there is no coincidence with the identification mark of the culture instrument used in the past, the process proceeds to step S24 to allow the start of the culture operation. If there is something that matches the identification mark of the culture instrument used in the past, the process proceeds to step S25, where a warning that the culture instrument is defective is output to the display unit 70, and the process is performed without starting the culture operation. finish.

  According to the present embodiment, it is possible to avoid a possibility that a culture instrument used for cell culture of a patient is erroneously reused for cell culture of another patient. Further, if the cell culture is carried out as it is, ignoring the warning, and some trouble occurs in the cultured cells, it can be estimated with high accuracy that the location of the problem lies in the culture instrument.

  Although not shown, the counter unit 68 can count the number of cell cultures every time the series of culture operations shown in FIG. According to this, it is possible to check whether an inappropriate culture instrument has been used or reused for another patient's cell culture by associating the number of culture instruments such as an incubator with the number of counters. In addition, there is a secondary effect that it can be used as an index for periodic maintenance of the apparatus.

  Moreover, in the above description, any method may be used for identifying the character image stamped on the culture apparatus or identifying the bar code with a barcode. For example, it may be a generally used IC tag, and is not particularly limited. In the case of sterilization with γ rays, an IC tag may be damaged due to the influence of radiation, so an image or a barcode is preferable as in this embodiment.

It is detail drawing of the incubator of one Embodiment used for the automatic culture apparatus of this invention. It is a whole block diagram of one Embodiment of the automatic culture apparatus of this invention. It is a block diagram which concerns on the characteristic part of the automatic culture apparatus of this invention. It is the 1 of the operation | movement flowchart of an example of an automatic culture apparatus. It is the 2 of the operation | movement flowchart of an example of an automatic culture apparatus. It is a flowchart of one Embodiment which concerns on the process of the characteristic part of this invention. It is a figure which shows an example of the record of the cultured cell of this invention. It is a flowchart of other embodiment which concerns on the process of the characteristic part of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Incubator 21, 40 Tube 22-24, 41, 42 Bag 35-37 Air filter 46 Image acquisition means

Claims (5)

  A plurality of culture devices including an incubator used for cell culture, reading means for reading an identification mark attached to at least one of the culture devices, and the identification mark read by the reading means is associated with information on cultured cells And an automatic culture apparatus comprising storage means for storing the information.   A plurality of culture devices including an incubator used for cell culture, a reading unit for reading an identification mark attached to at least one of the culture devices, and a registration in which the identification mark read by the reading unit is stored in advance An automatic culturing apparatus comprising: determination means for determining whether or not the identification mark is matched and outputting a warning if not matched.   A plurality of culture devices including an incubator used for cell culture; reading means for reading an identification mark attached to at least one of the culture devices; and storage means for storing the identification mark read by the reading means; Determination means for determining whether or not the same identification mark as the identification mark newly read by the reading means is already stored in the storage means, and the determination means includes the newly read identification mark. An automatic culture apparatus that warns that the same identification mark as the mark is already stored in the storage means.   A plurality of culture devices including an incubator used for cell culture, control means for controlling the cell culture by operating the culture devices in accordance with a culture procedure, and reading means for reading an identification mark attached to at least one of the culture devices And determination means for determining whether or not the identification mark read by the reading means is stored in advance and matches the registered identification mark, and the determination means includes the identification mark as the registered identification mark. An automatic culture apparatus that outputs a cell culture start permission command to the control means only when they match. In the automatic culture apparatus according to any one of claims 1 to 4,
An automatic culture apparatus comprising storage means for storing the cumulative number of automatic cell cultures based on the identification mark read by the reading means.

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