EP3794102A1 - Dispositif et procédé de culture de cellules dans le mode d'agitation - Google Patents

Dispositif et procédé de culture de cellules dans le mode d'agitation

Info

Publication number
EP3794102A1
EP3794102A1 EP19724379.3A EP19724379A EP3794102A1 EP 3794102 A1 EP3794102 A1 EP 3794102A1 EP 19724379 A EP19724379 A EP 19724379A EP 3794102 A1 EP3794102 A1 EP 3794102A1
Authority
EP
European Patent Office
Prior art keywords
gas exchange
opening
shaking
gas
gas phase
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.)
Pending
Application number
EP19724379.3A
Other languages
German (de)
English (en)
Inventor
David Frank
Konrad Herzog
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Aquila Biolabs GmbH
Original Assignee
Aquila Biolabs GmbH
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 Aquila Biolabs GmbH filed Critical Aquila Biolabs GmbH
Publication of EP3794102A1 publication Critical patent/EP3794102A1/fr
Pending legal-status Critical Current

Links

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
    • 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
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/24—Gas permeable parts
    • 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
    • C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
    • C12M27/16—Vibrating; Shaking; Tilting
    • 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
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/34—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of gas
    • 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
    • C12M41/42—Means for regulation, monitoring, measurement or control, e.g. flow regulation of agitation speed

Definitions

  • the invention relates to a device for cultivating cells in the shaking operation and a method for their application. It is particularly applicable to high oxygenation culture processes or to cultures with high requirements for establishing a gas phase equilibrium with the environment.
  • the invention finds advantageous application in cultivation processes with passive fumigation, the mixing of which is achieved by shaking, but especially not exclusively in the field of shake flask cultivations.
  • the cultivation of cells takes place in a variety of devices, which make the storage and mixing of the culture liquid 2 and the setting of cultivation conditions. Of particular importance is the composition of the gas phase, from which the culture fluid 2 is supplied with oxygen, or exchanged over the physiologically relevant gases such as CO2.
  • the maintenance of a particular gas phase composition is critical to the success of any cultivation process. Thus, e.g. only at a sufficiently high oxygen concentration in the gas phase, that oxygen partial pressure in the culture fluid 2, which is necessary to allow completely aerobic metabolic pathways. Due to the low solubility of oxygen in aqueous phases and the continuous oxygen consumption of breathing cells, a device for culturing cells must provide suitable means for continuous gas exchange 10.
  • this gas exchange 10 must take place as possible to maintain sterility in order to prevent contamination of the cultivation process with foreign cells.
  • the gas phase in the culture apparatus becomes frequent and varied Sterility requirement via a sterile barrier 5 separated from the outer gas phase.
  • While actively gassed cultivation devices such as stirred tank fermenters, sterile-filtered gas mixtures are actively introduced into the culture fluid 2 (eg by means of compressed air as a bubble column), the gas exchange 10 in passively fumigated cultivation devices such as shake flasks takes place only via slow diffusion and convection between the head space 3 of the cultivation device and its ambient gas phase 6
  • passively fumigated cultivation devices such as shake flasks
  • oxygen limitations which slow down the cultivation process and adversely affect the formation of undesired metabolites or fermentation products.
  • This effect is enhanced in some applications by additions or additions to the culturing apparatus which hinder the diffusion and convection both between and within the headspace gas phase 3 of the cultivation apparatus and the ambient gas phase 6, in particular sensors and probes as well as feeding and sampling systems.
  • Passively fumigated devices for cultivating cells are fundamentally based on the concept of a liquid container with variously closable filling opening upwards, via which the gas exchange 10 with the ambient gas phase 6 takes place, so that the fumigation and the handling of the Culture liquid 2 (filling, sampling, etc.) carried out through the same opening.
  • the mixing of such cultivation devices takes place via the movement of the device in interaction with the inertia and friction of the liquid and gas phases located in the device.
  • Widely used devices which implement this concept are in particular a wide variety of culture flasks and bottles as well as multi-well plates.
  • the mixing takes place in particular on rocking, tumbling or orbital shakers, which can also be designed as incubators, in addition to the mixing also defined temperature, humidity and ambient gas phase conditions.
  • a disadvantage of the passive fumigation culturing devices described above is the location and orientation of the device opening over which the gas exchange 10 takes place.
  • the shaking motion induced movement of the liquid 2 in the culture device causes gas flow within the headspace 3 of the device. Due to the device geometry (especially in piston type devices), the distance between the gas movement causing liquid 2 and the device opening and the orientation of the opening surface parallel to the device bottom and thus parallel to the main movement surface of the liquid is the movement and thus the local mixing of the gas phase in the headspace 3 just before the device opening very weak and also almost free of pressure gradient with significant vertical proportion with respect to the opening area, so that both the gas exchange 10 within the near-open headspace 3 and the gas exchange 10 via the device opening with the ambient gas phase 6 predominantly by diffusion without appreciable Pressure gradient must occur and is therefore slow.
  • W0001987007293A1 describes a shaker flask with gas exchange enhancing agents. While this also solves the problem of poor mixing of the gas phase within the headspace 3, the above-described disadvantageous position, alignment and execution of the gas exchange / filling opening remains.
  • DE000004415444C2 and EP000000905229A2 describe devices for detecting and monitoring the physiological state of microbial cultures by means of the oxygen transfer rate. Also disclosed are methods and apparatus for the active gassing of shake flasks as a cultivation device, the fumigation of the device taking place via separate gas exchange openings 4 opened at the top.
  • the active gassing solves the problem of slow gas exchange 10 both within the near-open headspace 3 and through the device openings with the ambient gas phase 6, since the active gassing results in efficient convective gas exchange 10.
  • the active fumigation disadvantageously increases the complexity and manageability of the entire cultivation device, since the shaking flasks must be equipped with hoses, pumps, valves, etc.
  • the object underlying the invention is achieved by a device according to claim 1 and a method of using the device according to claim 10; preferred embodiments will be apparent from the dependent claims and the description.
  • the gas exchange 10 designates the transition of gaseous substances between two phases, in particular between the ambient gas phase 6 and the headspace gas phase 3, and between the headspace gas phase 3 and the culture liquid 2.
  • Gas exchange 10 takes place by the movement of the substances to be exchanged, in particular by convection or diffusion.
  • active gas exchange 10 this takes place by means of dedicated gas conveying devices and methods, in particular by means of pumps or overpressure systems, wherein the gas to be exchanged is usually conducted through pipelines or hoses into the device or culture fluid 2.
  • the active gas exchange 10 can be controlled directly, for example, by changing the volume flow via the pump speed.
  • passive gas exchange 10 this takes place without dedicated gas delivery devices and methods, but by utilizing given conditions, such as ambient pressure and construction and movement of the device for cultivating cells.
  • the passive gas exchange 10 is not directly controllable, but only by changing the given conditions, but especially not by changing the temperature, pressure or composition of the ambient gas phase 6, as well as by altering the structure or movement of the cell culturing device ,
  • the head space 3 comprises the inner volume of the device, which is not filled by culture fluid 2.
  • the headspace gas phase 3 both together or separately, reference is made as headspace with headspace gas phase 3.
  • the device according to the invention is located in an environment with ambient gas phase 6, referred to both together or separately as the environment with ambient gas phase 6.
  • the composition of the ambient gas phase 6 either corresponds to the natural ambient air or is selectively defined, for example by gas mixing stations.
  • the use of the device according to the invention takes place in the shaking operation, but in particular not exclusively by rocking, tumbling or orbital shaking.
  • the shaking operation can take place both continuously and intermittently.
  • the device has a relative velocity along a velocity vector, which is called here the motion vector 9, at least to the environment with ambient gas phase 6.
  • the motion vector 9 may change over time, but not exclusively with regard to direction and magnitude.
  • the device flows around the gas, so depending on the point considered the device changes the pressure on the device and a local ambient pressure 8 for each point of the device flows around, which adjusts to the Shake movement and the motion vector 9 can change.
  • the difference or the pressure gradient between headspace pressure 7 and ambient pressure 8 can cause a convective or diffusive pressure compensation via at least one gas exchange 10. All this applies both to total pressures and to the partial pressures of individual components of a phase.
  • Culture fluids 2 liquids or liquid phases in the sense of the invention are pure or mixed substances which are not gases and have fluidic properties. Liquids in the sense of the invention are therefore not in particular exclusively liquid pure substances, solutions, emulsions, dispersions, sludges, suspensions and foams. Culture fluids 2 consist in particular but not exclusively of nutrient medium and cells.
  • a gas exchange opening 4 is an opening in the wall 1 of the device, which primarily serves the purpose of gas exchange 10. It is characterized by an opening area which results in the smallest possible area suitable for closing the opening.
  • the opening area may be curved and may be described at any point by at least one normal vector.
  • the orientation of an opening or opening area to the motion vector 9 of the shaking motion is described by the angle a, which results as the intersection angle between the motion vector b (9) and a normal vector n on the opening area
  • the angle a may differ from point to point, since the normal vectors of a curved surface are not all parallel.
  • the opening area of a gas exchange opening 4 and a motion vector 9 are considered to be non-parallel if, for at least one point on the opening area of the considered gas exchange opening 4, the angle a is greater than 0 °.
  • a sterile barrier 5 is a gas-permeable device, which is used in particular for the prevention, reduction or complete suppression of the penetration of unwanted cells, viruses or other contaminants into the interior of the device according to the invention by at least one gas exchange opening 4 or further opening 11.
  • Sterile barriers 5 according to the invention allow at least one gas exchange 10 between the head space with headspace gas phase 3 and the environment with ambient gas phase 6, in particular via diffusion and / or convection.
  • Sterile barriers 5 according to the invention are in particular but not exclusively sterile filters, porous membranes (eg PTFE, cellulose, hydrophilic or hydrophobic, etc.), wadding plugs or cushions and open-pore foams of silicone, polyurethane or other plastics.
  • Sterilbarrieren 5 in the context of the invention are connected to the wall 1 of the device according to the invention, but not exclusively by direct bonding or welding, and indirectly via suitable closure systems with screw or snap closure or other positive or material connections.
  • the object is achieved by a device for cultivating cells in the shaking operation, wherein the claimed device has at least one gas exchange opening 4 whose opening surface is aligned non-parallel to the motion vector 9 of the shaking motion.
  • This arrangement leads, caused by the shaking and the associated flow around the device according to the invention by the ambient gas phase 6 outside and the headspace gas phase 3 inside, to form a depending on the shaking movement changing pressure gradient between the headspace gas phase 3 and the ambient gas phase 6, which advantageously gas exchange between headspace 3 and ambient gas phase 6 and the gas mixing within the headspace 3 influenced.
  • this pressure gradient causes a compensating flow which, depending on the direction of the pressure gradient, allows gas to flow from the environment 6 into the headspace 3 of the device or to escape from the headspace 3 of the device into the device Environment 6 leads.
  • periodic shaking movements thus results in a breath-like oscillating gas movement between the headspace 3 and environment 6, which on the one hand allows a much more efficient gas exchange 10 than the known from the prior art passively fumigant cultivating devices and on the other hand improves the mixing within the headspace 3.
  • the pressure gradient between the environment 6 and the head space 3 forms, in particular, along the motion vector 9 of the device according to the invention in the shaking mode.
  • the pressure gradient increases for the same total area of the gas exchange opening 4 with increasing angle a between the opening area and the motion vector 9 from 0 ° to 90 °.
  • the achievable pressure gradient is maximum when the motion vector 9 is orthogonal to the opening area and minimal when the motion vector 9 is parallel to the opening area, the latter arrangement corresponding to the current state of the art.
  • the device is advantageously used in a method according to the invention for the cultivation of cells in the shaking mode.
  • a device according to the invention is filled with culture liquid 2.
  • a headspace gas phase 3 is established in the headspace.
  • the filled device according to the invention is positioned on or in a shaker and thus within an environment with ambient gas phase 6.
  • cells are now cultivated in the apparatus according to the invention, the shaking operation being characterized in that at least one movement vector 9 of the shaking movement runs non-parallel to the opening surface of at least one gas exchange opening 4.
  • the inventive method ensures that the device according to the invention is used under such conditions, under which it comes to the formation of at least one depending on the shaking movement changing pressure gradient between the headspace gas phase 3 of the device according to the invention and its ambient gas phase 6 and advantageous compared to the prior art, the gas exchange 10 between headspace gas phase 3 and ambient gas phase 6 and the gas mixing within the headspace gas phase 3 can be improved.
  • the angle a between the opening area of at least one gas exchange opening 4 and at least one movement vector 990 ° applies to the angle a between the opening area of at least one gas exchange opening 4 and at least one motion vector 960 ° ⁇ a ⁇ 90 ° or 45 ° ⁇ a ⁇ 90 ° or 45 ° ⁇ a ⁇ 60 "or 30 ° ⁇ a ⁇ 90 ° or 30 ° ⁇ a ⁇ 60 ° or 30 ° ⁇ a ⁇ 45 ° or 0 ° ⁇ a or 15 ° ⁇ a or 30 ° ⁇ a or 45 ° ⁇ a or 60 ° ⁇ a.
  • the device comprises a plurality of gas exchange openings 4, in particular but not exclusively two, three, four, five or six.
  • the device is surrounded by a plurality of gas exchange openings 4, so that for almost every state of the shaking for at least one of these gas exchange openings 4 at least one of the above conditions for the angle a between the opening area and motion vector 9 applies.
  • each gas exchange opening 4 of the device according to the invention with a sterile barrier 5 is closed, which hinders the penetration of foreign cells or completely prevented, however, is permeable to the gas phase components to be exchanged.
  • at least one sterile barrier 5 allows the gas exchange 10 via convection and diffusion.
  • the gas exchange openings 4 are arranged in a common plane in order to ensure the stability of the device during Scblinins.
  • the device is autoclavable or pre-sterilized.
  • the sterile barriers 5 can be changed.
  • the device in addition to at least one gas exchange opening 4, the device comprises at least one further opening 11, which is suitable but not exclusively for filling and emptying the device, for sampling, for attaching probes and sensors or for attaching Feeding- and dispensing ,
  • further openings 11 comprise a closure system for attaching external devices or closures 12, in particular but not exclusively closure systems with plug-in, latching, form-fitting or screwing mechanism.
  • one or more gas exchange openings 4 are deposited from the wall 1 of the device according to the invention and / or stocky with an inner splash guard 13 to the leakage of liquid due to Scblini- or Flandhabungsdochen and the contact of liquid with any existing sterile barriers. 5 to prevent.
  • one or more gas exchange ports 4 are mounted in a flea above the bottom of the device that is at least twice as high as the fleas of the unshaken culture fluid 2 above the bottom during conventional filling, but especially not only when the device is filled with 5% to 20% of their nominal volume.
  • the gas exchange 10 between the headspace gas phase 3 and the ambient gas phase 6 is unidirectional, in other embodiments bidirectional.
  • selective sterile barriers 5 are used, which allow only one or more specific gas phase components for the gas exchange 10 via the respective gas exchange opening 4.
  • the wall 1 is made of glass. In other embodiments of the device according to the invention, the wall 1 is made of plastic, but in particular not exclusively by means of injection or centrifugal casting.
  • Figure 1 a schematic representation of the device according to the invention.
  • FIG. 3 a representation of an embodiment of the invention as a shaking flask with circulating gas exchange openings 4th Figure 4, a schematic representation of the device according to the invention with connected via another opening 11 Feeding system.
  • FIG. 1 shows a schematic representation of the device according to the invention.
  • This comprises at least one wall 1, which encloses the interior of the device according to the invention.
  • the headspace gas phase 3 is in communication with the environment with ambient gas phase 6, so that in particular but not exclusively via diffusion and / or convection of gas phase components at least one Gas exchange 10 between the headspace gas phase 3 and the ambient gas phase 6 can take place.
  • Each gas exchange opening 4 may be closed by a gas-permeable sterile barrier 5.
  • the device according to the invention is mixed by occasional, periodic or continuous shaking, wherein the shaking motion is characterized at any time via at least one motion vector 9.
  • the device also comprises at least one further opening 11, which is closed at least during the cultivation of cells, but in particular not exclusively with a suitable closure 12th
  • FIG. 2 shows a schematic representation of three embodiments of the device according to the invention as a shaking flask with different gas exchange openings 4.
  • A, B, C are each a side section (above the dashed line) and a top view (below the dashed line) given.
  • the embodiments in FIG. 2 A-C all include a further opening 11 for filling, emptying and sample pulling, feeding or for carrying out other interventions in the cultivation process, which is designed here as a classic piston opening and is closed with a lid as closure 12.
  • the embodiment of the device according to the invention as a shake flask comprises a comparatively large, upwardly tapered headspace with headspace gas phase 3 at typical nominal filling volumes of up to 30%.
  • This classic headspace geometry benefits in particular in orbital-shaken operation from the advantages of the passive gas exchange 10 according to the invention via gas exchange openings 4, their opening area is aligned non-parallel to the motion vector 9 of the shaking motion.
  • shaking flasks according to the prior art have only a single opening upwards, which is only poorly suited for gas exchange 10 because of their orientation, which is wholly or during the shaking operation, which is predominantly parallel to the shaking movement.
  • gas exchange 10 is somewhat weaker but still significantly better than passive gasification devices according to the current state of the art.
  • the gas exchange opening 4 is advantageously offset obliquely from the wall 1 of the device according to the invention, so that spray liquid and condensation liquid do not disadvantageously wet or close the sterile barrier 5, but instead flow back into the culture liquid 2.
  • FIG. 2C shows an embodiment of the device according to the invention as a shaking flask with a multiplicity of circulating gas exchange openings 4, which are all provided with a common sterile barrier 5 and thus represent a quasi-continuous gas exchange opening 4.
  • This arrangement is particularly advantageous in Orbital conveyel
  • the device according to the invention since they ensures that the gas exchange 10 favoring non-parallel arrangement between the motion vector 9 and the opening area of the gas exchange opening 4 is given at any time of the shaking.
  • the gas exchange openings 4 are not set down but integrated directly into the wall 1 so that the sterile barrier 5 rests directly on the wall 1 of the device and the angle a between the opening area of the gas exchange openings 4 and the motion vector 9 by the geometry of the wall 1 results.
  • This offers particular advantages in terms of manufacturing simplification of a device according to the invention, since here the sterile barrier 5 can be applied more easily, but especially not by gluing or ultrasonic welding to the wall 1. It is also advantageous that the non-discontinuation of the gas exchange openings 4 of the focus of the invention Device closer to the ground remains, which has a positive effect on the stability of the device and the maximum allowable shaking speed, especially with rapid shaking.
  • the embodiment of Fig. 2C comprises a splash guard 13 which is mounted between the gas exchange port 4 and the culture liquid 2 to retain spouting liquid.
  • a splash protection curvature 13 can also be advantageously used above the gas exchange opening 4, in particular to prevent wetting of the sterile barrier 5 with condensate flowing out of the upper device part.
  • the settling of at least one gas exchange opening 4 is combined with at least one of the aforementioned splash guard bumps 13.
  • FIG. 3 A selected implementation of the device according to the invention from FIG. 2C is shown in FIG. 3 as a perspective partial section.
  • the device is designed here as a classical shaker flask comprising a wall 1, in which above the culture liquid 2 radially surrounding several gas exchange openings 4 are introduced, so that an entire, quasi-continuous gas exchange opening 4 for gas exchange 10 between Kopfraumgasphase 3 and ambient gas phase 6 results only through narrow connecting struts 14 is interrupted. Also circumferentially covered from the outside a contiguous sterile barrier 5 (shown here cut) all gas exchange openings 4. Below this extends annularly a splash guard arch 13. The device also has a further opening 11 with closure 12th
  • the embodiment of the device according to the invention shown in FIG. 3 is particularly suitable for being manufactured as a disposable product made of plastic and delivered presterilized.
  • Figure 4 shows a schematic representation of the device according to the invention with connected via a further opening 11 Feeding system.
  • the device is designed here as a classical shaker flask, comprising a wall 1, to which above the culture liquid 2 a plurality of gas exchange openings 4 in a remote version for gas exchange 10 between Kopfraumgasphase 3 and ambient gas phase 6 are mounted such that at least one time of shaking the angle a between Motion vector 9 and the opening area of at least one gas exchange opening 4 is greater than 0 ° and motion vector 9 and opening area of at least one gas exchange opening 4 are aligned non-parallel to each other.
  • a gas-permeable sterile barrier 5 covers each gas exchange openings 4.
  • the device has a further opening 11, which is closed during operation of the apparatus according to the invention for cultivating cells by a closure with integrated Feeding reservoir 15 and on which a dosing system 16 is mounted.
  • This advantageously allows the addition of substrates and nutrients during the cultivation of cells, in particular but not exclusively as a fed-batch process, so that the apparatus shown here as a result of the invention improved gas exchange 10th is able to carry out cell cultivation processes without oxygen limitation even with passive fumigation, which otherwise would only be possible in significantly more complex, actively fumigated stirred tank fermenters.

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  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Clinical Laboratory Science (AREA)
  • Analytical Chemistry (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

La présente invention a pour but de fournir un dispositif au moyen duquel la culture de cellules peut se produire lors d'une fumigation passive dans le mode d'agitation, avec un échange gazeux optimal (10) entre une phase gazeuse d'espace de tête (3) et une phase gazeuse ambiante (6), ainsi qu'un mélange gazeux optimal au sein de l'espace de tête (3). À cet effet, l'invention concerne un dispositif destiné à la culture de cellules dans le mode d'agitation, le dispositif concerné possédant au moins une ouverture d'échange gazeux (4), dont la surface d'ouverture n'est pas parallèle au vecteur de mouvement (9) du mouvement d'agitation. Du fait du mouvement d'agitation et de l'écoulement lié à ce dernier du dispositif selon l'invention par la phase gazeuse ambiante (6) à l'extérieur et la phase gazeuse d'espace de tête (3) à l'intérieur, ledit agencement mène à la formation d'un gradient de pression variant selon le mouvement d'agitation entre la phase gazeuse d'espace de tête (3) et la phase gazeuse ambiante (6), qui agit avantageusement sur l'échange gazeux entre la phase gazeuse d'espace de tête (3) et la phase gazeuse ambiante (6) ainsi que sur le mélange gazeux au sein de l'espace de tête (3).
EP19724379.3A 2018-05-16 2019-05-08 Dispositif et procédé de culture de cellules dans le mode d'agitation Pending EP3794102A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102018003910.3A DE102018003910B8 (de) 2018-05-16 2018-05-16 Vorrichtung und Verfahren zur Kultivierung von Zellen im Schüttelbetrieb
PCT/EP2019/061768 WO2019219465A1 (fr) 2018-05-16 2019-05-08 Dispositif et procédé de culture de cellules dans le mode d'agitation

Publications (1)

Publication Number Publication Date
EP3794102A1 true EP3794102A1 (fr) 2021-03-24

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EP19724379.3A Pending EP3794102A1 (fr) 2018-05-16 2019-05-08 Dispositif et procédé de culture de cellules dans le mode d'agitation

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US (1) US20210189311A1 (fr)
EP (1) EP3794102A1 (fr)
DE (1) DE102018003910B8 (fr)
WO (1) WO2019219465A1 (fr)

Family Cites Families (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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
US4665035A (en) 1986-05-27 1987-05-12 Josephino Tunac Fermentation apparatus and systems for the cultivation of microorganisms and other biological entities
US5100801A (en) * 1989-01-26 1992-03-31 Biocontrol Systems, Inc. Device for sequential microbial enrichment in a single apparatus
DE4415444C2 (de) 1993-05-12 2002-10-31 Jochen Buechs Automatisches Meßsystem zur sterilen on-line Bestimmung der Sauerstofftransferrate (OTR) in Schüttelkolben
EP0905229B1 (fr) 1997-09-01 2004-04-28 Toyota Gosei Co., Ltd. Procédé et dispositif pour déterminer et surveiller l'état physiologique de cultures microbiennes
US8409854B2 (en) * 2001-07-31 2013-04-02 Sartorius Stedim Biotech Gmbh Bioreactor provided with equipment with flexible walls
TWI325401B (en) * 2007-12-17 2010-06-01 Duen Gang Mou Vessel structure
US8696778B2 (en) * 2009-11-05 2014-04-15 Todd A. McAdams Self-contained breathing closure and container
CN202430229U (zh) * 2011-12-22 2012-09-12 上海康岱生物医药技术股份有限公司 一种大容量细胞培养转瓶
DE102014001284B3 (de) * 2014-02-01 2015-01-22 aquila biolabs GmbH Verfahren, Vorrichtung und System zur automatisierten Bestimmung optischer Dichten oder der Veränderung optischer Dichten von Reaktionsgemischen in geschüttelten Reaktoren
PL3132020T3 (pl) * 2014-04-17 2020-06-29 Corning Inc. Naczynie do hodowli komórek i jego zastosowanie
CN207079243U (zh) * 2017-08-16 2018-03-09 苏州米迪生物技术有限公司 扰流装置

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DE102018003910B3 (de) 2019-07-04
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