WO2019051486A1 - Bioréacteur à grande échelle - Google Patents

Bioréacteur à grande échelle Download PDF

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Publication number
WO2019051486A1
WO2019051486A1 PCT/US2018/050467 US2018050467W WO2019051486A1 WO 2019051486 A1 WO2019051486 A1 WO 2019051486A1 US 2018050467 W US2018050467 W US 2018050467W WO 2019051486 A1 WO2019051486 A1 WO 2019051486A1
Authority
WO
WIPO (PCT)
Prior art keywords
liquid
culture
cells
flow
sub
Prior art date
Application number
PCT/US2018/050467
Other languages
English (en)
Inventor
Zongsen Wang
Wing Lau
Faribourz Payvandi
Peter Materna
Original Assignee
3D Biotek, Llc
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 3D Biotek, Llc filed Critical 3D Biotek, Llc
Priority to CN201880059387.4A priority Critical patent/CN111132595A/zh
Priority to EP18852871.5A priority patent/EP3681365A4/fr
Publication of WO2019051486A1 publication Critical patent/WO2019051486A1/fr

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
    • C12M23/58—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
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/14—Scaffolds; Matrices
    • 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
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/10—Perfusion
    • 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
    • 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
    • C12M33/08—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by vibration
    • 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
    • C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
    • C12M37/02—Filters
    • 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

Definitions

  • Figure 1 A is a three-dimensional perspective view of a culture chamber mounted above a reservoir.
  • the liquid pump 450 (which may be a peristaltic pump) to the showerhead 410.
  • the moat 160 there is a certain volume of the moat 160 as defined by space from the bottom surface of the moat 160 to the top of the overflow weir wall 140 that defines the moat 160. If the liquid pump 450 has been operating in the forward direction for some time, it can be expected that the tubing is full of liquid. It also is typical that the liquid level in the moat 160 is fairly low, i.e., close to the bottom of the moat 160. It may be desirable that when the direction of flow in the tubing is reversed, the liquid pump 450 may operate so as to introduce gas entering the tubing from the showerhead 410.
  • liquid flow could be operated in an oscillating manner.
  • the liquid flow direction could change repeatedly, and the volume of liquid displaced during any one oscillation could be relatively small, as could the distance that a given segment of liquid moves through the scaffold during oscillation.
  • Such a situation could be produced, using a peristaltic pump, if the rotor of the peristaltic pump rotates back and forth alternating its direction of rotation.
  • Such oscillation could be sinusoidal but does not have to be.
  • the liquid pump 450 can be operated in alternate directions for a small amount of volume displacement while the scaffold region is still fully submerged in liquid. This can cause alternating up and down flow of liquid past the scaffolds, which may be appropriate for dislodging cells from the scaffolds. It would also be possible to combine, in some sequence, the just-described alternating flow with the just-described pulsatile flow. For example, some reverse-direction flow of liquid could be followed by forward-direction flow of liquid in a relatively strong velocity or flowrate, which could be followed by a period of more gentle liquid flow. Any of this could be simultaneous with externally imposed vibration as may be desired.
  • the frequency of the oscillation of the flow could be different, even significantly different, from the frequency of vibration; alternatively, if desired, the frequency of the oscillation of the flow could be the same as, or almost the same as, the frequency of vibration. In the latter situation, the vibration and the flow oscillation could be adjusted to be in-phase with each other, in a way such that accelerations experienced by the cells due to vibration could reinforce forces experienced by the cells due to liquid motion.
  • the progress of the harvesting process can be estimated by observing the flow resistance (or the change in flow resistance) of the scaffold as a function of time during the harvesting process.
  • the flow resistance of the scaffold can be characterized in generally the same way that has been described herein in connection with estimating the degree of cell growth (approach to confluency) during the culturing process, by using pumping-related information.
  • the scaffold would have a relatively large flow resistance, which would be reflected in the pressure drop.
  • the flow resistance can be determined from a calculation using the liquid flowrate and the pressure drop.
  • the flow resistance of the scaffold would likely be smaller. This information could be used to determine how long the harvesting process should continue. There is potential for the harvesting process to damage cells, so it is advantageous that the harvesting process not continue longer than necessary. Similarly, this information could be used to adjust what technique is used at a given time during the harvesting process.
  • Bioreactors can be monitored for any of various process parameters associated with their operation, including but not limited to: pH of the culture medium; temperature; concentration of glucose in the culture medium; concentration of lactate in the culture medium; concentration of dissolved oxygen in the culture medium; concentration of carbon dioxide in the atmosphere above the liquid; numbers or confluence of cells growing on substrates. It is also possible that any of these could be used as a parameter to control a feedback loop that would adjust a process parameter to achieve a desired result.
  • each culture chamber there could be provided a plurality of sub -reservoirs each having a culture chamber associated therewith. It is possible that for each culture chamber there can be a dedicated fluid flow circuit that moves liquid culture medium past the scaffolds during culturing. Such circuit can have individual control of fluid flowrate, such as by an individually controlled liquid pump 450. In response to the conditions as indicated by a sensor, it is possible to adjust any one or more of the following during either cell culturing or cell harvesting: volumetric flowrate of liquid; duration of liquid flow; direction of liquid flow.
  • harvesting operations could be done differently for different culture chambers, and may be done responsive to sensed values of any of the described parameters. For example, harvesting operations do not have to be performed simultaneously for all of the culture chambers; rather, harvesting operations could be performed when a determination is made that for that particular culture chamber, an appropriate level of progress toward confluence has been reached. Also, the duration of harvesting operations does not have to be identical for all of the culture chambers 100.
  • liquid culture medium can be removed and replaced with harvesting liquid.
  • an overflow weir wall 140 defining a moat 160 with an exit at a lower elevation than the top of the overflow weir wall 140, such that when in operation, there is a trapped volume of gas above the liquid that is inside the culture chamber 100.
  • the presence of a trapped volume of gas is not essential, and as an alternative it is also possible to operate a culture chamber 100 in a mode in which the interior of the culture chamber 100 is completely filled with liquid.
  • embodiments of the invention are a closed system, easier and less expensive to operate, requires less maintenance and is more automated than currently available system. For some applications it is desired to harvest and make use of the cultured cells themselves. However, it is not always necessary to harvest cells from a bioreactor. There are some other applications in which the secretions of the cells are of interest, rather than the cells themselves.
  • saline solution could be Phosphate Buffered Saline.
  • the term pressure measuring device is intended to encompass a pressure transducer, a pressure transmitter, and any other suitable device for measuring pressure.

Landscapes

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

Abstract

Dans un mode de réalisation de l'invention, il peut y avoir un bioréacteur ayant des échafaudages tissulaires et ayant un milieu de culture perfusé à travers celui-ci. Il peut y avoir de multiples chambres de culture indépendantes et des réservoirs ou sous-réservoirs. Des capteurs peuvent permettre de commander individuellement des conditions dans diverses chambres de culture, et diverses chambres de culture peuvent être actionnées différemment ou pour différentes durées. Il est possible de déduire le nombre de cellules ou la progression vers la confluence à partir de la résistance aux fluides de l'échafaudage, sur la base du débit et de la chute de pression. La récolte peut comprendre n'importe quelle combinaison ou séquence de : l'exposition au réactif de récolte ; une vibration ; un écoulement de liquide qui est constant, pulsatile ou oscillant ; le passage d'une interface gaz-liquide à travers l'échafaudage. Des vibrations et un écoulement peuvent être appliqués de manière à se renforcer l'un l'autre.
PCT/US2018/050467 2017-09-11 2018-09-11 Bioréacteur à grande échelle WO2019051486A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CN201880059387.4A CN111132595A (zh) 2017-09-11 2018-09-11 大型生物反应器
EP18852871.5A EP3681365A4 (fr) 2017-09-11 2018-09-11 Bioréacteur à grande échelle

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US201762556646P 2017-09-11 2017-09-11
US62/556,646 2017-09-11
US201862636039P 2018-02-27 2018-02-27
US62/636,039 2018-02-27

Publications (1)

Publication Number Publication Date
WO2019051486A1 true WO2019051486A1 (fr) 2019-03-14

Family

ID=65634631

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2018/050467 WO2019051486A1 (fr) 2017-09-11 2018-09-11 Bioréacteur à grande échelle

Country Status (3)

Country Link
EP (1) EP3681365A4 (fr)
CN (1) CN111132595A (fr)
WO (1) WO2019051486A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022202733A1 (fr) * 2021-03-26 2022-09-29 Terumo Kabushiki Kaisha Système de culture cellulaire
WO2022202732A1 (fr) * 2021-03-26 2022-09-29 Terumo Kabushiki Kaisha Système de culture cellulaire

Citations (7)

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US20110236970A1 (en) * 2008-08-01 2011-09-29 Smart Biosystems Aps Chamber of a bioreactor platform
US20130177972A1 (en) * 2009-11-17 2013-07-11 David Green Bioreactors, systems, and methods for producing and/or analyzing organs
US20140030805A1 (en) * 2011-04-15 2014-01-30 Pluristem Ltd. Methods and systems for harvesting cells
WO2014102730A1 (fr) * 2012-12-28 2014-07-03 Universidade Federal De Minas Gerais - Ufmg Chambre de perfusion de culture tridimensionnelle pour le génie tissulaire
WO2015061907A1 (fr) * 2013-10-30 2015-05-07 Miklas Jason Dispositifs et procédés de culture de tissu tridimensionnel
EP2151491B1 (fr) * 2008-08-06 2015-10-28 Associacion for the Advancement of Tissue Engineering and Cell Based Technologies & Therapies (A4TEC) Bioreacterur multi-chambre avec perfusion bidirectionnel integre dans un systeme de culture pour des strategies d'ingenierie de tissus

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US20110236970A1 (en) * 2008-08-01 2011-09-29 Smart Biosystems Aps Chamber of a bioreactor platform
EP2151491B1 (fr) * 2008-08-06 2015-10-28 Associacion for the Advancement of Tissue Engineering and Cell Based Technologies & Therapies (A4TEC) Bioreacterur multi-chambre avec perfusion bidirectionnel integre dans un systeme de culture pour des strategies d'ingenierie de tissus
US20130177972A1 (en) * 2009-11-17 2013-07-11 David Green Bioreactors, systems, and methods for producing and/or analyzing organs
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WO2014102730A1 (fr) * 2012-12-28 2014-07-03 Universidade Federal De Minas Gerais - Ufmg Chambre de perfusion de culture tridimensionnelle pour le génie tissulaire
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022202733A1 (fr) * 2021-03-26 2022-09-29 Terumo Kabushiki Kaisha Système de culture cellulaire
WO2022202732A1 (fr) * 2021-03-26 2022-09-29 Terumo Kabushiki Kaisha Système de culture cellulaire

Also Published As

Publication number Publication date
EP3681365A4 (fr) 2021-06-16
EP3681365A1 (fr) 2020-07-22
CN111132595A (zh) 2020-05-08

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