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WO1997031128A1 - Appareil et procedes de culture de cellules de mammiferes - Google Patents

Appareil et procedes de culture de cellules de mammiferes Download PDF

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Publication number
WO1997031128A1
WO1997031128A1 PCT/US1997/002704 US9702704W WO9731128A1 WO 1997031128 A1 WO1997031128 A1 WO 1997031128A1 US 9702704 W US9702704 W US 9702704W WO 9731128 A1 WO9731128 A1 WO 9731128A1
Authority
WO
WIPO (PCT)
Prior art keywords
cell culture
media
culture media
pump
tube
Prior art date
Application number
PCT/US1997/002704
Other languages
English (en)
Inventor
Glenn F. Spaulding
Original Assignee
Vivorx Pharmaceuticals, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Vivorx Pharmaceuticals, Inc. filed Critical Vivorx Pharmaceuticals, Inc.
Priority to AU21331/97A priority Critical patent/AU2133197A/en
Publication of WO1997031128A1 publication Critical patent/WO1997031128A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/26—Means for regulation, monitoring, measurement or control, e.g. flow regulation of pH
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/08—Flask, bottle or test tube
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/48—Automatic or computerized control
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q3/00—Condition responsive control processes

Definitions

  • the present invention relates to a novel apparatus and process related thereto, for monitoring and feeding cell cultures, particularly mammalian and other extremely sensitive cell cultures.
  • T-flas s The most widely practiced processes for mammalian cell culture utilize T-flas s.
  • cells are obtained from some source, for example a human biopsy or commercial cell line, and placed into cell culture media.
  • a sterile T-flask is prepared and media containing the cells is deposited into the T-flask.
  • the T-flask is then placed in a C0 2 incubator and the cells are allowed to grow.
  • the T-flask is observed for colorimetric changes in the media caused by a pH indicator dye that is commonly added to the media. When the dye changes color, indicating an undesirable pH change, the T-flask is removed from the incubator and placed under a sterile hood for feeding.
  • Feeding requires the cap of the T-flask to be removed and placed aside in a sterile location, and the cap of the vessel containing the fresh cell culture media to be removed and placed in a sterile location.
  • the T- flask is tipped upright to move the depleted media to the pipetteable space at the bottom of the T-flask, which causes the non-adherent cells to be washed to the bottom of the T-flask along with the depleted media.
  • a sterile pipette is placed into the T-flask and the depleted media, along with the non-adherent cells, is removed and discarded.
  • a new sterilized pipette is obtained and fresh cell culture media is transferred into the T-flask, the T-flask cap replaced, and the T-flask returned to its original position in the incubator.
  • U.S. Patent No. 4,680,267 assigned to New Brunswick Scientific, relates to an interactive 4-gas instrumentation system for control of dissolved oxygen (D.O.) and pH.
  • the system disclosed in '267 is large, complex and expensive, and does not readily adapt to standard T-flasks.
  • Monitoring is invasive.
  • monitoring according to the present invention is non- invasive.
  • the present invention enables simple and low cost monitoring and is readily optimized for use with standard T-flasks, without the need for modifications.
  • Biosciences, Inc. relates to cell culture wells and continuous radial perfusion.
  • the system disclosed in '069 is large, complex and expensive, and does not readily adapt to standard T-flasks.
  • the present invention enables simple and low cost monitoring and is readily optimized for use with standard T-flasks, without the need for modifications.
  • the present invention provides an integrated solution to the above-described problems in the art.
  • optical pH monitoring enables non-invasive and quantitative monitoring; media replenishment takes place directly in the cell culture container (i.e., the cell culture base is established after adherent and non-adherent cells have settled to the bottom of the cell culture container) ; only partial media replenishment is required, thus lessening the dilution of growth factors; and media changes occur at user supplied pH thresholds while the cell culture container remains in the incubator.
  • invention apparatus and methods lower labor costs for maintaining cell cultures and reduce the amount of cell culture media required, thus providing a lower cost process. Moreover, the invention apparatus and methods enable improved outcomes for a variety of cell cultures.
  • the present invention provides a simple, cost- effective way to feed and maintain a cell culture without non-adherent cell loss, with reduced risk of contamination, while controlling the pH within any desired limits. Additionally, the cells are not exposed to the harsh temperature, air, and pH extremes, nor the large growth factor fluctuations, typically associated with conventional T-flask cell culture techniques.
  • the apparatus is small and of simple design such that it can be attached to a variety of other cell culture systems, for example a horizontally rotating bioreactor or shaking bioreactor. In the case of moving bioreactors, the pumping action can be accomplished by direct coupling to the agitating mechanism, thereby eliminating an extra motor. A variety of tubing types and diameters can be used with the apparatus.
  • Figure 1 is a block diagram of an apparatus according to the invention, including operative connections between the various components thereof.
  • Figure 2 is a schematic layout of an optical pH sensor, demonstrating the relationship between the pH sensor and the tube.
  • the present invention provides a cell culture apparatus comprising: a) a cell culture container shaped to hold cell culture media; b) a tube comprising a first portion and a second portion, wherein said first portion is within said cell culture container, and said second portion is outside said cell culture container; c) a pH sensor capable of operative communication with cell culture media in which the cell culture is growing; d) a pump, wherein said pump is in operative communication with said second portion of said tube; e) a source of fresh cell culture media in fluid communication with said second portion of said tube; f) a receptacle for waste cell culture media in fluid communication with said second portion of said tube; and g) a control device, wherein said control device is in operative communication with said pump and said pH sensor, said control device controlling said pump in response to said pH sensor.
  • the present invention provides a method to grow cells under controlled pH conditions, said method comprising: a) inoculating cell culture media present within a cell culture apparatus according to invention with a cell culture; and b) monitoring and adjusting the pH of said cell culture media as the cells grow.
  • the present invention provides a method for the replenishment of cell culture media in a cell culture container without disturbing said cell culture media, said method comprising: a) monitoring the pH of a portion of said cell culture media with a pH sensor by pumping a portion of said cell culture media out of said cell culture container into a tube, wherein said tube comprises a first portion within said cell culture container in fluid communication with said cell culture media and a second portion outside said cell culture container, wherein said second portion of said tube is in operative communication with said pH sensor, a pump, a source of fresh cell culture media, and a receptacle for waste cell culture media; and b) controlling further action of said pump in response to said pH sensor, wherein i) if the pH of said cell culture media is within the target parameters, said pump is caused to return said portion of said cell culture media to said cell culture container, or ii) if the pH of said cell culture media is not within the target parameters, said pump is caused to transfer said cell culture media from said cell culture container to said recept
  • the present invention enables pH monitoring and feeding of a cell culture in a non-invasive manner.
  • non-invasive it is meant that items absent at the time of inoculating the cell culture are not introduced or brought into contact with the cell culture in order to monitor pH, feed the cells, or the like.
  • the present invention also enables the cell culture to be monitored or fed without disturbing the cell culture media.
  • “Disturbing" the cell culture media means to subject the cell culture media to undue physical agitation, temperature changes, exposure to air, and the like.
  • Prior art cell culture techniques require disturbing the cell culture media in order to monitor or feed it, while the practice of the present invention can readily eliminate such disturbances, and thereby promote better growth of the cell culture.
  • a block diagram of the cell culture apparatus 10 and its functional connections is shown.
  • the cell culture grows by feeding on the cell culture media in a cell culture container 12.
  • a first portion 16 of a tube 14 is within the cell culture container 12, and a second portion 18 of the tube 14 is placed in operative communication with a pH sensor 20, a pump 22, a source of fresh cell culture media 24, and a receptacle for waste cell culture media 26.
  • the control device 28 causes the pump 22 to draw cell culture media from the cell culture container 12 into the tube 14 to a position where the pH sensor 20 can measure the pH of the cell culture media.
  • the pH determination is compared to the pH target parameters that have been previously set in the control device 28 by the operator. If the pH of the cell culture media is not within the pH target parameters, the depleted cell culture media is withdrawn from the cell culture container 12 and replaced with fresh cell culture media from the fresh media source 24.
  • the cell culture container 12 is a standard T-flask.
  • the invention embraces any type of container that may be used to culture cells, including non-moving bottles, petri dishes, and the like.
  • the tube 14 is a platinum cured silicone tube with an inner diameter of 1/16" and a wall thickness of 1/16".
  • a platinum cured silicone tube with an inner diameter of 1/16" and a wall thickness of 1/16".
  • Useful materials for the tubing include all varieties of silicone rubber, all types of plastics, and the like.
  • the size of the tubing can range from an inner diameter of about 0.001 inches to about 2 inches, while the tubing wall thickness can range from about 0.001 inches to about 0.5 inches.
  • the tube 14 typically comprises a first portion 16 that is inside the cell culture container 12, and a second portion 18 that is outside the cell culture container 12.
  • the first portion 16 is passed through an opening in the cell culture container 12.
  • the first portion 16 typically lies on the base 30 of the cell culture container 12, with a hole 32 in the tube 14 approximately one tubing wall diameter above the base 30 9 of the cell culture container 12. By locating the hole 32 above the base 30, it is not possible to withdraw all of the cell culture media from the cell culture container 12. Residual cell culture media will always remain in the cell culture container 12 to protect the cell culture from direct exposure to air. In contrast, in prior art cell culture practices, the cells are commonly exposed to air.
  • the depth of the remaining media will be approximately equal to the distance between the base 30 of the cell culture container 12 and the hole 32 in the tube 14.
  • the depth of the cell culture media is important both for protecting cells and for determining growth factor dilutions.
  • the tube 14 is in direct contact with the base 30, while the hole 32 is offset from the base 30. In this manner, the distance between the base 30 and the hole 32 is directly related to the thickness of the wall of the tube 14.
  • the depth of the media that remains in the cell culture container 12 may be adjusted by merely selecting a tube 14 having a different wall thickness.
  • the pH sensor 20 is preferably an optical pH sensor. In practice, almost any pH sensor 20 that can measure pH without increasing the risk of infecting or otherwise harming the cell culture may be used in the present invention. Other examples of useful pH sensors 20 include FET, electrodes, and the like.
  • the pH sensor 20 is in operative communication with the cell culture media when the media has been pumped into the tube 14 to a point where the tube 14 is within or next to the pH sensor 20.
  • the tube 14 is routed from the cell culture container 12 through a notch in the optical pH sensor 20. This allows the pH sensor to measure the pH of the media without the media leaving the sterile environment inside the tube 14, and without introducing any outside agents into that sterile environment. It is in this manner that non-invasive optical measurement of pH is readily achieved.
  • the cell culture media contains a pH sensitive agent that changes color as the pH changes.
  • the pH sensitive agent is required if non- invasive pH monitoring is to be carried out.
  • suitable pH sensitive agents include Phenol Red, and any other compounds that change color at the appropriate pH range, and which are nontoxic to the cell culture.
  • Phenol Red transitions from red to orange as the pH transitions from pH 7.5 to pH 6.8.
  • a difference in the light absorbance occurs.
  • cell culture media is replenished according to the invention methods or using the invention apparatus.
  • the present invention requires the use of a pump 22 to move the cell culture media between the cell culture container 12, through the tube 14, and into waste receptacle 26 or out from the fresh media source 24. It is also very important that the pump 22 move the media in a relatively slow and easy fashion. This insures that non-adherent cells in the cell culture are not accidentally removed from the cell culture container 12 along with the depleted media.
  • the pump 22 moves the media at a rate of from about 0.1 ml/min to about 1.0 ml/min. Most preferably, the rate of movement is from about 0.25 ml/min to about 0.5 ml/min.
  • a peristaltic pump 22 is used for pumping media.
  • a peristaltic pump 22 is utilized because of its ability to provide suction, low cost, and precision control of media volume and dispensing/withdrawing rates. In practice, any pump 22 that is able to provide precise volume control, in combination with precise pumping rates, will be useful in the practice of the present invention.
  • the amount of media to withdraw, the media replacement volumes, and the pump 22 on/off times and rates are readily determined by those skilled in the art.
  • Media is suctioned from the cell culture container 12, through the tube 14, and past the pH sensor 20. If the media is within the pH target parameters, the media is pumped back into the cell culture container 12. If the pH is outside the pH target parameters set by the operator, the pump 22 continues to withdraw media. Once the cell culture container 12 has been emptied (except for the residual media) , the pump direction is reversed and fresh media is pumped into the cell culture container 12.
  • the tube 14 In order to dispose of depleted media, the tube 14 must be in fluid communication with the waste media receptacle 26.
  • the waste media receptacle 26 may be almost any type of container for holding the depleted media, or it may even be as simple as a sink or sewer into which the depleted media may flow for disposal purposes.
  • fresh media Once the depleted media has been removed from the cell culture container 12, fresh media must be supplied.
  • the fresh media source 24 may be virtually any container that is capable of holding sterile media suitable for growing the cells being cultured. Almost any size or shape of container that may be sterilized and then maintained in a sterile condition is useful.
  • a first check valve 34 and a second check valve 36 may be placed in-line in the tube 14.
  • the first check valve 34 is positioned between the waste receptacle 26 and the pump 22.
  • the second check valve 36 is positioned between the fresh media source 24 and the pump 22.
  • the flow directions of check valves 34 and 36 are opposite to each other. As such, when the pump 22 is drawing depleted media from the cell culture container 12, the first check valve 34 is open and the second check valve 36 is closed. When the pump 22 is drawing fresh media from the source of fresh media 24, the first check valve 34 is closed and the second check valve 36 is open.
  • a control device 28 is required.
  • the control device 28 ensures that pH measurements of the cell culture media are made at the desired times, and that the pump is activated to replenish the media when necessary.
  • the control device 28 causes the pump 22 to bring a sample of the cell culture media into operative communication with the pH sensor 20 every X hours, where X is about 0.1 hours to about 20 hours. In a preferred embodiment, X is about 1 hour to about 8 hours.
  • control device 28 is a microcomputer that has been preprogrammed with the necessary instructions to operate the apparatus.
  • the microcomputer may instead be a programmable device that is merely capable of being programmed with the necessary instructions.
  • Other embodiments of the control device 28, according to the present invention include a full- sized computer, a human operator, and the like.
  • the cell culture apparatus is controlled by the control device 28.
  • a data input device 38 and a visual display 40 that are part of the control device 28.
  • the visual display 40 is a standard LCD display.
  • the visual display 40 can also be a television screen, a computer monitor, and the like.
  • the data input device 38 is a standard button key pad.
  • the data input device 38 can also be a full keyboard, a modem or dedicated line (i.e., for connection to distant data input devices 38) , a voice- activated terminal, and the like.
  • the visual display 40 and data input device 38 are interfaced with the control device 28 by methods that are commonly used and known by those skilled in the art.
  • the operator sets the pH target parameters by pushing the keys on the button key pad. Once entered the control device 28 stores the information for later comparison with the pH determination made by the pH sensor 20. If the pH is outside the pH target parameters entered by the operator, the pump 22 is turned on for a preprogrammed duration that is based on the known volume of media in the cell culture container 12.
  • the depleted cell culture media is transferred from the cell culture container 12 into the waste media receptacle 26. Then the direction of the pump 22 is reversed for a preprogrammed duration during which time fresh cell culture media is transferred from the fresh media source 24 into the cell culture container 12.
  • first LED 50 and second LED 52 are both directed to first photosensor 54, through tube 14, then to second photosensor 56.
  • First photosensor 54 measures the light output from LEDs 50 and 52, and is used to compensate for light output variations, using methods known in the art.
  • Second photosensor 56 is used to measure the light output from LEDs 50 and 52 after some of the light has been absorbed by the media inside the tube 14.
  • the wavelength output from first LED 50 is in the green range, and for second LED 52 the wavelength output is in the blue range.
  • first LED 50 is turned on by the control device 28, and the signal outputs of photosensors 54 and 56 are registered by the control device 28 (these signal outputs correspond to the light energy from first LED 50 that impacts the photosensors 54 and 56) .
  • first LED 50 is turned off and second LED 52 is turned on.
  • the signal outputs of photosensors 54 and 56 are again registered by the control device 28.
  • the signal outputs registered from photosensors 54 and 56 under both conditions are compared to a look-up table preprogrammed in the control device 28 to determine the pH. Establishing a pH look-up table and programming it into the control device 28 are well known in the art.
  • the pH value derived from the look-up table is the measured pH of the cell culture media.
  • the control device 28 compares the measured pH to the target parameters previously set by the operator, and controls the actions of the other components of the cell culture apparatus 10 (especially the pump 22) on the basis of this comparison.
  • a platinum cured silicone tube (1/16" I.D., 1/16" wall) was placed through a hole in the cap of a standard 250 ml T-flask.
  • the 1/16" wall thickness of the tube insured that the proper volume of media remained in the T-flask during the media withdrawal process.
  • the opposite end of the tube was connected to a Y-connector, which was used to place the tube into operative communication with a waste receptacle and a fresh media source.
  • a first check valve which only allowed flow away from the T-flask was placed in-line in the leg of the tube connected to the waste receptacle.
  • a second check valve which only allowed flow toward the T-flask was placed in-line in the leg of the tube connected to the fresh media source.
  • the tube was placed in a notch in an optical pH sensor.
  • the pump was turned on by a microcomputer, and approximately 400 microliters of media was withdrawn from the T-flask (this being the volume required to bring the media into the tube and in operative communication with the optical pH sensor) .
  • the microcomputer compared data from the pH sensor to the operator supplied target parameters (pH 7.2 or greater) .
  • the pump When the media pH had dropped below pH 7.2, the pump was turned on by the microcomputer and 100 ml of media was withdrawn (or 2 times the normal media volume for a standard 250 ml T-flask) . Media removal was at a rate of 1.7 ml/min, which left non-adherent cells inside the cell culture container. After media removal, the microcomputer reversed the direction of the pump and infused 50 ml of fresh media from the fresh media source into the T-flask.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
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  • Health & Medical Sciences (AREA)
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  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

Appareil (10) de culture de cellules de mammifères et procédés permettant le réapprovisionnement d'un récipient de culture cellulaire (12) en milieu de culture cellulaire sans perturber le milieu de culture cellulaire déjà présent dans ledit récipient. Ledit appareil (10) de culture cellulaire est constitué d'un récipient de culture cellulaire (12) adapté pour contenir le milieu de culture cellulaire; d'un tube (14) comportant une première partie (16) située à l'intérieur du récipient (12) et une seconde partie (18) située à l'extérieur de celui-ci; d'un capteur de pH (20) apte à être mis en communication active avec le milieu de culture dans lequel se produit la croissance de la culture cellulaire; d'une pompe (22) mise en communication active avec ladite seconde partie (18) dudit tube (14); d'une source (24) de milieu de culture frais, cette source étant mise en communication fluidique avec ladite seconde partie (18) dudit tube (14); d'un réceptacle (26) recevant le milieu de culture cellulaire usé et se trouvant en communication fluidique avec ladite seconde partie (18) dudit tube (14); et d'un dispositif de commande (28) mis en communication active avec ladite pompe (22) et ledit capteur de pH (20), et destiné à commander ladite pompe (22) en réponse audit capteur de pH (20).
PCT/US1997/002704 1996-02-22 1997-02-21 Appareil et procedes de culture de cellules de mammiferes WO1997031128A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU21331/97A AU2133197A (en) 1996-02-22 1997-02-21 Apparatus and methods for culturing mammalian cells

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US1208696P 1996-02-22 1996-02-22
US60/012,086 1996-02-22

Publications (1)

Publication Number Publication Date
WO1997031128A1 true WO1997031128A1 (fr) 1997-08-28

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0870823A1 (fr) * 1997-03-07 1998-10-14 Istituto Trentino Di Cultura Dispositif de contrÔle de l'activité métabolique de cellules vivantes
DE10259251B4 (de) * 2001-12-19 2008-07-10 National Cheng Kung University Kultivierkammer an einem Mikroskopgestell
WO2009118015A3 (fr) * 2008-08-01 2010-03-04 Smart Biosystems Aps Orifice pour échantillons d'un système de culture cellulaire
EP2924110A4 (fr) * 2013-12-26 2016-03-23 Panasonic Ip Man Co Ltd Dispositif de culture cellulaire et procédé de culture cellulaire
CN109943483A (zh) * 2018-11-03 2019-06-28 宁波大学 基于实时补充技术的细胞培养装置及方法

Citations (6)

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Publication number Priority date Publication date Assignee Title
US3740321A (en) * 1970-04-15 1973-06-19 Smith Kline French Lab Cell propagator
US3875000A (en) * 1974-04-05 1975-04-01 Merck & Co Inc Cell culture apparatus
US3941662A (en) * 1971-06-09 1976-03-02 Max-Planck-Gesellschaft Zur Forderung Der Wissenschaften E.V. Apparatus for culturing cells
US3950227A (en) * 1970-12-21 1976-04-13 St. John's University Batch method of establishing and maintaining a controlled aerobic environment for a microbial culture
DE2657209A1 (de) * 1976-12-17 1978-06-22 Bender & Hobein Gmbh Biokulturverfahren und vorrichtung zu seiner ausuebung
US4812392A (en) * 1984-12-27 1989-03-14 Sumitomo Electric Industries, Ltd. Method and apparatus for incubating cells

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3740321A (en) * 1970-04-15 1973-06-19 Smith Kline French Lab Cell propagator
US3950227A (en) * 1970-12-21 1976-04-13 St. John's University Batch method of establishing and maintaining a controlled aerobic environment for a microbial culture
US3941662A (en) * 1971-06-09 1976-03-02 Max-Planck-Gesellschaft Zur Forderung Der Wissenschaften E.V. Apparatus for culturing cells
US3875000A (en) * 1974-04-05 1975-04-01 Merck & Co Inc Cell culture apparatus
DE2657209A1 (de) * 1976-12-17 1978-06-22 Bender & Hobein Gmbh Biokulturverfahren und vorrichtung zu seiner ausuebung
US4812392A (en) * 1984-12-27 1989-03-14 Sumitomo Electric Industries, Ltd. Method and apparatus for incubating cells

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0870823A1 (fr) * 1997-03-07 1998-10-14 Istituto Trentino Di Cultura Dispositif de contrÔle de l'activité métabolique de cellules vivantes
DE10259251B4 (de) * 2001-12-19 2008-07-10 National Cheng Kung University Kultivierkammer an einem Mikroskopgestell
WO2009118015A3 (fr) * 2008-08-01 2010-03-04 Smart Biosystems Aps Orifice pour échantillons d'un système de culture cellulaire
EP2924110A4 (fr) * 2013-12-26 2016-03-23 Panasonic Ip Man Co Ltd Dispositif de culture cellulaire et procédé de culture cellulaire
US9650599B2 (en) 2013-12-26 2017-05-16 Panasonic Intellectual Property Management Co., Ltd. Apparatus for culturing cells and method for culturing cells
CN109943483A (zh) * 2018-11-03 2019-06-28 宁波大学 基于实时补充技术的细胞培养装置及方法

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