EP2640824A1 - Dispositif de passage des cellules - Google Patents
Dispositif de passage des cellulesInfo
- Publication number
- EP2640824A1 EP2640824A1 EP10782551.5A EP10782551A EP2640824A1 EP 2640824 A1 EP2640824 A1 EP 2640824A1 EP 10782551 A EP10782551 A EP 10782551A EP 2640824 A1 EP2640824 A1 EP 2640824A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- culture vessel
- suspension
- riser
- cells
- cell
- Prior art date
- Legal status (The legal status 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 status listed.)
- Withdrawn
Links
- 238000000034 method Methods 0.000 claims abstract description 29
- 238000004113 cell culture Methods 0.000 claims abstract description 15
- 239000000725 suspension Substances 0.000 claims description 57
- 239000007788 liquid Substances 0.000 claims description 47
- 239000012530 fluid Substances 0.000 claims description 26
- 238000007599 discharging Methods 0.000 claims description 4
- 238000004891 communication Methods 0.000 claims description 2
- 238000012545 processing Methods 0.000 abstract description 2
- 238000012258 culturing Methods 0.000 abstract 1
- 235000015097 nutrients Nutrition 0.000 description 12
- 239000000243 solution Substances 0.000 description 12
- 239000007853 buffer solution Substances 0.000 description 10
- 102000004190 Enzymes Human genes 0.000 description 9
- 108090000790 Enzymes Proteins 0.000 description 9
- 229940088598 enzyme Drugs 0.000 description 9
- 239000002609 medium Substances 0.000 description 9
- 239000007789 gas Substances 0.000 description 7
- 230000005484 gravity Effects 0.000 description 7
- 230000001464 adherent effect Effects 0.000 description 6
- 238000000605 extraction Methods 0.000 description 6
- 238000011109 contamination Methods 0.000 description 5
- 239000006285 cell suspension Substances 0.000 description 4
- 238000004140 cleaning Methods 0.000 description 4
- 238000007639 printing Methods 0.000 description 4
- 239000002699 waste material Substances 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 230000004663 cell proliferation Effects 0.000 description 3
- 238000012432 intermediate storage Methods 0.000 description 3
- 238000007789 sealing Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 238000012546 transfer Methods 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- KFSLWBXXFJQRDL-UHFFFAOYSA-N Peracetic acid Chemical compound CC(=O)OO KFSLWBXXFJQRDL-UHFFFAOYSA-N 0.000 description 2
- 102000004142 Trypsin Human genes 0.000 description 2
- 108090000631 Trypsin Proteins 0.000 description 2
- 239000003570 air Substances 0.000 description 2
- 238000011166 aliquoting Methods 0.000 description 2
- 239000006143 cell culture medium Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 238000005204 segregation Methods 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000012588 trypsin Substances 0.000 description 2
- 238000002604 ultrasonography Methods 0.000 description 2
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 239000012080 ambient air Substances 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000012864 cross contamination Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 229910001882 dioxygen Inorganic materials 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 229940125532 enzyme inhibitor Drugs 0.000 description 1
- 239000002532 enzyme inhibitor Substances 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 238000005187 foaming Methods 0.000 description 1
- 230000002706 hydrostatic effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000010943 off-gassing Methods 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 1
- 229940024999 proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 239000012192 staining solution Substances 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
-
- 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
- C12M23/58—Reaction vessels connected in series or in parallel
-
- 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
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/12—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by pressure
Definitions
- the invention is in the field of automatic cultivation and further processing of biological cells and tissues and relates to an apparatus and a method for the automatic passage of biological cells or tissues during cell cultivation by means of pressurization.
- Biological cells are capable of dividing; Individual cells can form cell aggregates and, if appropriate, entire tissue layers over a specific cultivation period. For effective cell proliferation, this process is repeated iteratively. For example, individual cells are cultured into a tissue matrix, the tissue matrix is dissolved and the cells are separated. The isolated cells are then re-cultivated in each case to form cell clusters or tissues.
- This process is referred to as "passage.”
- the transfer of cells from a first cell culture vessel into one or more cell culture vessels is required in known manual processes by preparing a homogenous cell suspension in the first culture vessel, aspirating the cells Cell suspension by means of a pipette and aliquoting of the cell suspension sucked into the pipette into the further cell culture vessels
- Known solutions for the automated passage are based on a replication of these manual processes by technical means
- CONFIRMATION COPY and replaced the hand of the laboratory staff by a suitable robot kinematics.
- the conventional procedure of manual passage is described using the example of the passage of adherent cells: in a first step, the culture vessel containing the adherent cells is opened, in a second step supernatant over the cells, which as a rule consists of spent nutrient medium, aspirated, in a further step, the adhering to the bottom of the culture vessel cells are rinsed by optionally multiple overlays with buffer solution. In a further step, enzyme solution (trypsin) is added to dissolve the tissue assembly and detach the cells from the bottom of the vessel, and as a rule incubate with agitation.
- trypsin enzyme solution
- the enzyme activity is stopped by enzyme inhibitor to be dosed.
- the detached cells and cell aggregates or pieces of tissue are suspended in nutrient medium.
- the suspension is removed by suction from the culture vessel.
- the cell density number of cells per unit volume in the suspension is determined.
- the cell suspension is distributed to other cell culture vessels (aliquoted) and the cell culture vessels are sealed.
- the known automation of the known manual process requires a high expenditure on equipment in order to simulate the work steps and actions of the laboratory staff. At the same time, the known automation does not overcome the major disadvantages of manual passage. These are above all the risk of contamination, uncontrolled mechanical stress on the biological cells during the pipetting process, which is mainly characterized by alternating application of negative pressure and overpressure (sucking up, pumping out), uncontrollable flow conditions and multiple changes in the flow direction. Furthermore, inaccuracies in the aliquoting of the cells due to segregation of the suspension occur during the residence of the suspension in the pipette.
- the invention is based on the technical problem of providing a device for the automatic passage of biological tissue or cells (cell passage device) which substantially or completely obviates the above-described disadvantages of the prior art overcomes.
- the invention solves this technical problem, in particular by completely turning away from solution principles known from manual passage methods. Especially the use of a pipette is avoided.
- the invention provides a cell passage device for automatically passing a suspension of biological tissues or cells or liquid from a first culture vessel into at least one or more further culture vessels.
- This device contains according to the invention at least one removal unit for receiving and discharging the suspension or liquid from the first culture vessel.
- the removal unit has at least one riser for receiving the suspension or liquid and at least one pressure tube for pressurizing the first culture vessel.
- riser and pressure tube of the extraction unit can be inserted into the first cell culture vessel and the removal unit can complete the cell culture vessel pressure-tight.
- the removal unit is specially designed so that in the inserted state, the riser tube dips into the suspension or liquid present in the first cell culture vessel and the pressure tube is located above the liquid level.
- the cell passage device has at least one metering unit for metering the suspension or liquid into one or more of the further culture vessels.
- the dosing unit is specially designed to transfer the suspension or liquid removed by means of the integral withdrawal unit into one or more of the further culture vessels, especially to aliquot it distributed into a plurality of further culture vessels.
- the cell passage device according to the invention has at least one reservoir for liquid (liquid reservoir).
- a plurality of liquid reservoirs are provided.
- the liquid reservoirs are specially designed to receive the media required for the passage, such as cell culture medium (nutrient medium), buffer or washing liquid, enzyme solution and optionally resuspension liquid and sterilization liquid.
- the quantities required for the passage process can be metered automatically from the reservoirs.
- the cell passage device according to the invention has a printing original. This is specially designed to provide the pressure required for carrying out the passage process according to the invention.
- the artwork is optionally connectable to one or more of the foregoing elements to facilitate liquid transport (suspension transport) along the pressure gradient formed, optionally against gravity (hydrostatic pressure).
- a further element of the cell passage device is a fluidic block which has a plurality of fluid channels and a plurality of valves associated with the fluid channels.
- the fluid channels with the above-described elements especially with extraction unit, metering unit, at least one liquid reservoir and the artwork in fluid communication.
- the fluidic block, fluid channels and valves are specially designed to selectively sequentially fluidly connect one or more of the protruding elements to at least one other of the protruding elements to carry out the passage, preferably to selectively pressurize and, therefore, conditional to allow a targeted liquid transport within the cell passage device.
- the fluidic block is preferably further characterized in that the fluidic channels in the fluidic block are oriented substantially horizontally.
- the fluid channels are oriented substantially perpendicular to the gravity vector.
- the cell passage device according to the invention is specially designed to minimize the residence time of a suspension in fluid lines running substantially parallel to the gravity vector. Thus, the influence of gravity-induced segregation of the suspension on the distribution of the suspended cells or tissues can be prevented or reduced.
- the invention now provides that by pressurizing the first culture vessel via the at least one pressure tube of the integral sampling element, the liquid or suspension in the first cell culture vessel can be removed via the at least one riser of the integral sampling unit.
- biological cells or tissue parts thus suspended can be carefully removed from the first cell culture vessel in a controlled manner and transferred into the dosing unit via the fluidic block, which can preferably additionally serve as a buffer for the suspension.
- the fluidic block which can preferably additionally serve as a buffer for the suspension.
- the invention further provides for the media stored in the liquid reservoirs to be sequentially and selectively pressurized. tion of the liquid reservoir via the fluidic block and the integral withdrawal unit in the first culture vessel to dose.
- the dosage of enzyme solution into the first culture vessel is thus made possible in order to bring there adherent cells in suspension in order to remove them from the culture vessel.
- the solution according to the invention is based on the fact that liquid or suspension is not aspirated out of the culture vessel, temporarily stored in a pipette and then dispensed in metered quantities. Instead, liquid or suspension are pressed by pressurization via the pressure tube of the integral extraction unit according to the invention into a riser tube of the extraction unit according to the invention immersed in the liquid or suspension, and possibly guided without further intermediate storage and while maintaining the flow direction.
- the suspension can thus be removed from the first culture vessel and "pressed" into one or more of the further culture vessels in a subsequent step while maintaining the direction of flow, preferably without intermediate storage, into the downstream dosing unit for metering the suspension Underpressure and overpressure, an intermediate storage with associated residence times with separation risk and the disadvantageous multiple reversal of the flow direction are avoided.
- the invention also preferably provides that at substantially constant pressure, the amount of liquid is controlled by the opening duration of the valves in the fluidic block, taking into account the specific flow rate.
- the fluidic block is used together with the valves as a central control unit to allow the flow of liquid and suspension in the device.
- the control of the valves is preferably carried out programmatically.
- the program control allows an adjustment of the dosing quantities, the predetermination of the number of aliquots in connection with the number of additional culture vessels provided in each case as well as the preselection of additional washing, sterilization and / or calibration steps.
- riser and pressure tube are formed together as an integral unit in the form of a double needle or multiple needle.
- This is preferably pierced by an elastic septum, which is formed on the culture vessel, wherein preferably the septum pressure-tightly closes the culture vessel in the region of the pierced needle.
- additional means for pressure-tight sealing of the culture vessel are provided on the removal unit. These can be designed in a manner known per se as sealing lips, surface or labyrinth seals, which come into sealing engagement with the culture vessel in a special embodiment.
- the pressure is 0.5 to about 2 bar.
- the gas required for pressurization can be provided sterile and contamination-free.
- the gas is ambient air.
- the system allows the use of gas or gas compositions favorable for cell cultivation, for example, carbon dioxide-enriched oxygen gas (95% O 2 /5% CO 2 ). This allows the use of cell-friendly physical buffer systems in the cell culture media.
- the pressure can be provided in the simplest case via a connected gas cylinder; There is no need for a mechanical pump, which further reduces the risk of contamination and simplifies maintenance.
- the cell passage device makes it possible to carry out the passage on the basis of a purely unidirectional flow of the suspension.
- the danger of the formation of residuals in known pipetting processes due to the necessary flow reversal there is thus avoided.
- the device according to the invention allows, in a special embodiment, a substantially continuous transport / flow of the suspension in the passage, whereby in particular the residence time of the suspension within the removal unit can be minimized.
- the extraction unit can therefore optionally be guided parallel to the gravity vector (vertical). Any necessary retention of the suspension in the device according to the invention takes place only in the region of the horizontal fluid channels in the fluid block. Because of the arrangement according to the invention of the fluid channels, essentially perpendicular to the gravity vector, the separation of the suspension caused by gravity, that is to say an unintentional lowering of the cells in the fluid channel, is avoided.
- the device according to the invention avoids, in a particular embodiment, any kinematic processes within the system formed by the removal unit, dosing unit, liquid reservoir, printing original and fluidic block. As a result, dead volumes can advantageously be minimized.
- Usable media for the realization of the passage are in particular buffer solutions or nutrient-enriched nutrient media, which are known per se.
- Enzyme solutions, in particular trypsin, which may be used in conjunction with EDTA, may be used for the cell separation and detachment of adherent cells to produce a suspension.
- cleaning media for the lines, vessels and valves can be provided via a liquid reservoir according to the invention, in order to free the entire device from cells and other contaminants, for example in preparation for a passage process or subsequently.
- Ethanol, propanol, peracetic acid and / or hydrogen peroxide can be used in particular as cleaning media.
- the cross-section of the fluid channels does not exceed a certain size to secure formation Menisken and to prevent mixing of air or gas with liquid or suspension.
- an abrupt change in the cross section of the fluidic channels that is to say particularly abrupt transitions, corners and edges, is further avoided by avoiding through flow cavities and negative pressure with the risk of outgassing the liquid or suspension. This can prevent foaming in the system.
- a further aspect of the invention is a method for the automatic passage of cells or tissues in suspension from a first culture vessel into one or more further culture vessels using the cell passage device according to the invention described herein.
- the method has at least the following steps: In a first step, the removal unit containing at least one riser and at least one pressure tube is introduced into the first culture vessel and the culture vessel is closed pressure-tight, preferably via the removal unit. In a further step, the interior of the culture vessel is pressurized with the pressure tube, whereby liquid or suspension in the culture vessel is removed via the at least one riser, following the pressure gradient.
- the method according to the invention is further characterized in that the liquid or suspension discharged via the riser pipe is transferred via a metering unit connected to the riser pipe into at least one further culture vessel.
- the pressurization of the culture vessel via the pressure tube the only driving force for discharging the liquid or suspension from the culture vessel, and preferably for transferring the liquid or suspension, via the metering unit in at least one further culture vessel.
- the method further provides that for transferring the liquid or suspension pension from the first culture vessel into the at least one further culture vessel an, at least temporary, fluid connection between the riser and the metering unit is produced.
- this fluid connection is always flowed through in the same flow direction during the automatic passage of the liquid or suspension.
- Figure 1 shows an embodiment of the extraction unit (1 10) according to the invention with a riser (1 12) for receiving the suspension or other liquid and a pressure tube (1 14) for pressurizing, which are introduced into a culture vessel (200).
- riser (1 12) and pressure tube (114) are formed as an integral double needle, which is inserted in the region of the septum (202) of the first culture vessel (200).
- the culture vessel (200) is sealed pressure-tight with respect to the environment.
- Figure 2 shows the overall structure of a particular embodiment of the cell passage device (100) according to the invention with removal unit (110), dosing unit (120), liquid reservoirs (131, 132, 133, 134) and print template (140).
- the reservoir (131) preferably contains buffer solution
- the reservoir (132) preferably contains enzyme solution
- the reservoir (133) preferably contains nutrient medium
- the reservoir (134) preferably contains cleaning fluid.
- the dosing unit (120) has a single needle (122) which can be selectively brought into the further culture vessels (210) in order to maintain the suspension there. pension or liquid to dose.
- the valves (154) and the fluid connections between the valves and the aforementioned elements are arranged in an integral fluidic block (150) (not shown in the schematic diagram). In the illustrated embodiment, the valves are electromagnetically actuated.
- the control takes place via a central computer by means of hardware / software implemented program.
- waste container (160) and pressure pump (170) with pressure reducer (172) are provided.
- Example: Automated Passage The first culture vessel (200) containing the cells or tissues to be passed is provided.
- the removal unit (110) with riser (112) and pressure tube (114) is introduced into the culture vessel (200), wherein the riser (112) is positioned below the liquid level, as close as possible to the bottom of the culture vessel (200).
- the pressure tube (114) is located in the culture vessel above the liquid level.
- the printing original (140) is connected to the pressure tube (114) in order to pressurize the interior of the culture vessel (200) with overpressure.
- the liquid in the culture vessel (200) deviates via the riser (112) and is thus conveyed out of the culture vessel (200) in the direction of the fluidic block (150).
- nutrient medium supernatant is removed from the culture vessel (200).
- the nutrient medium removed from the culture vessel (200) is removed from the system via the fluidic block (150).
- the riser (112) via the fluidic block (150) with an optional waste container (160) is brought into connection and the pressurization of the culture vessel (200) the nutrient medium from the culture vessel (200) via the riser (112) and the fluidic block (150) in the optional waste container (160) out.
- buffer solution is added to wash the cells or tissues remaining in the culture vessel (200).
- the liquid reservoir is brought into contact with buffer solution (131) by switching at least one valve within the fluidic block (150) to the printing master (140) in order to meter buffer solution from the reservoir (131).
- the buffer solution metered from the reservoir (131) is metered into the culture vessel via the riser (112) or optionally via another feed tube of the removal unit (113, not shown).
- the added buffer solution is removed by pressurization of the culture vessel (200) via the pressure tube (114) via the riser (112) as described above and preferably transferred to the optional waste container (160).
- the process of washing is optionally repeated once or several times.
- enzyme solution is then removed from reservoir (132) by actuation of the corresponding valves in fluidic block (150) via riser (112) or optionally via the additional supply tube (113) for detachment of adherent cells to prepare the suspension ) into the culture vessel (200) and incubate the cells in the culture vessel (200) with enzyme solution. After the incubation period and, if appropriate, by assisting the detachment process by mechanical vibrations (ultrasound, etc.), by adding a solution inhibiting the enzyme activity (analogous for the above-mentioned addition of a buffer solution) stopped.
- nutrient medium in particular from reservoir (133) is metered into the culture vessel (200) in the above-described manner by actuation of at least one corresponding valve in the fluidic block (150) via riser (112) or optional additional feed tube (113).
- the detached cells are suspended (resuspended) in the dosed nutrient medium.
- the formation of a substantially homogeneous suspension by additional mechanical measures (ultrasound, etc.) in the culture vessel (200) is supported.
- the suspension obtained is then fed via the riser (112) into the fluidic block (150) and via the fluidic block (150) into the metering unit (120) by pressurizing the culture vessel (200) via the pressure tube (114) .
- Targeted control of the opening times of the valves preferably makes it possible to meter the suspension onto the dosing unit (120) and thus to aliquot the suspension to a plurality of culture vessels (210).
- the device is rinsed and cleaned by flushing with cleaning fluid from Reservoir (134) after passage, in order to avoid cross-contamination between individual passages.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Sustainable Development (AREA)
- Clinical Laboratory Science (AREA)
- Molecular Biology (AREA)
- Cell Biology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/EP2010/006995 WO2012065618A1 (fr) | 2010-11-17 | 2010-11-17 | Dispositif de passage des cellules |
Publications (1)
Publication Number | Publication Date |
---|---|
EP2640824A1 true EP2640824A1 (fr) | 2013-09-25 |
Family
ID=44352186
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP10782551.5A Withdrawn EP2640824A1 (fr) | 2010-11-17 | 2010-11-17 | Dispositif de passage des cellules |
Country Status (3)
Country | Link |
---|---|
US (1) | US20130236955A1 (fr) |
EP (1) | EP2640824A1 (fr) |
WO (1) | WO2012065618A1 (fr) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2014039082A1 (fr) * | 2012-09-04 | 2014-03-13 | Becton, Dickinson And Company | Pré-concentration bactérienne et technique de détection |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB1471865A (en) * | 1973-04-06 | 1977-04-27 | Unilever Ltd | Dispensers for use in bacteriology and similar biochemical analysis |
US6599484B1 (en) * | 2000-05-12 | 2003-07-29 | Cti, Inc. | Apparatus for processing radionuclides |
EP1869476A2 (fr) * | 2005-03-22 | 2007-12-26 | Irm Llc | Dispositifs et systemes de profilage de composes, et procedes associes |
US8263389B2 (en) * | 2007-01-12 | 2012-09-11 | Biorep Technologies, Inc. | Perifusion device |
JP5125210B2 (ja) * | 2007-04-27 | 2013-01-23 | 株式会社Ihi | 培養細胞サンプリング方法及びその装置 |
-
2010
- 2010-11-17 EP EP10782551.5A patent/EP2640824A1/fr not_active Withdrawn
- 2010-11-17 US US13/988,174 patent/US20130236955A1/en not_active Abandoned
- 2010-11-17 WO PCT/EP2010/006995 patent/WO2012065618A1/fr active Application Filing
Non-Patent Citations (2)
Title |
---|
None * |
See also references of WO2012065618A1 * |
Also Published As
Publication number | Publication date |
---|---|
WO2012065618A1 (fr) | 2012-05-24 |
US20130236955A1 (en) | 2013-09-12 |
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