GB2053704A - A flask for use in stirring particles in suspension such as microcarriers for anchorage-dependent living cells in a liquid culture medium - Google Patents

A flask for use in stirring particles in suspension such as microcarriers for anchorage-dependent living cells in a liquid culture medium Download PDF

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Publication number
GB2053704A
GB2053704A GB7924810A GB7924810A GB2053704A GB 2053704 A GB2053704 A GB 2053704A GB 7924810 A GB7924810 A GB 7924810A GB 7924810 A GB7924810 A GB 7924810A GB 2053704 A GB2053704 A GB 2053704A
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United Kingdom
Prior art keywords
flask
microcarriers
suspension
culture medium
anchorage
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Withdrawn
Application number
GB7924810A
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.)
Techne Cambridge Ltd
Original Assignee
Techne Cambridge Ltd
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 Techne Cambridge Ltd filed Critical Techne Cambridge Ltd
Priority to GB7924810A priority Critical patent/GB2053704A/en
Priority to GB8023215A priority patent/GB2054397B/en
Publication of GB2053704A publication Critical patent/GB2053704A/en
Priority to US06/277,403 priority patent/US4382685A/en
Withdrawn legal-status Critical Current

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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
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/08—Flask, bottle or test tube
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F27/00—Mixers with rotary stirring devices in fixed receptacles; Kneaders
    • B01F27/80—Mixers with rotary stirring devices in fixed receptacles; Kneaders with stirrers rotating about a substantially vertical axis
    • B01F27/95—Mixers with rotary stirring devices in fixed receptacles; Kneaders with stirrers rotating about a substantially vertical axis with stirrers having planetary motion, i.e. rotating about their own axis and about a sun axis
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F35/00—Accessories for mixers; Auxiliary operations or auxiliary devices; Parts or details of general application
    • B01F35/50—Mixing receptacles
    • 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
    • C12M27/02—Stirrer or mobile mixing elements
    • 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
    • C12M27/18—Flow directing inserts
    • C12M27/20—Baffles; Ribs; Ribbons; Auger vanes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F27/00—Mixers with rotary stirring devices in fixed receptacles; Kneaders
    • B01F27/23—Mixers with rotary stirring devices in fixed receptacles; Kneaders characterised by the orientation or disposition of the rotor axis
    • B01F27/232—Mixers with rotary stirring devices in fixed receptacles; Kneaders characterised by the orientation or disposition of the rotor axis with two or more rotation axes
    • B01F27/2324—Mixers with rotary stirring devices in fixed receptacles; Kneaders characterised by the orientation or disposition of the rotor axis with two or more rotation axes planetary

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Biomedical Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Clinical Laboratory Science (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The flask bottom is formed with an upwardly directed conical portion 3 delimited by an annular trough 5. The flask is designed so that the speed of revolution of the stirrer 1 stirring the medium in the flask can be low enough to avoid damage to the cells being swept around in the medium while at the same time maintaining the microcarriers in uniform suspension and obviating the risk of accumulation of the microcarriers at the centre or edge of the flask bottom. <IMAGE>

Description

SPECIFICATION A flask for use in stirring particles in suspension such as microcarriers for anchorage-dependent living cells in a liquid culture medium Field of the invention The present invention relates to a flask for use in stirring particles in suspension and particularly to a flask which may be employed for use in stirring micro-carriers or beads within a liquid culture suspension to enable the growth of anchoragedependent living cells thereon.
Background of the invention The growth of living cells in a stirred liquid culture medium is well established and is important both in the research and industrial fields.
Mammalian cell growth such as in the case of cancer cells can be generated in a stirred suspension culture without the need for the presence of an attachment surface. However certain mammalian cell types exist which can only grow in the suspension by being attached to a solid surface, and such cell types are termed anchorage-dependent.
In recent years methods for growing anchoragedependent cells in suspension cultures have been developed using microcarriers, see for example U.S.
Patent Specification Serial No. 4036693. The use of such microcarriers is more economic relative to other known methods and provides environmental continuity of growth since the cells can be grown in a controlled environment which can be regulated to provide constant optimal cell growth.
Normal methods for cell growth in a liquid culture medium containing microcarriers involves the necessity to maintain the microcarriers in uniform suspension while avoiding damage to the growing anchorage-dependent cells.
The accepted method of stirring non-anchorage dependent cells has been by a horizontal revolving permanent magnet encased in PTFE suspended by some form of bearing about 10 mm above the bottom of the flask containing the suspension. Such a device is described in U.S. Patent Specification Serial No. 3572651. The device disclosed therein is, however, ineffective for stirring microcarriers in suspension since they tend to accumulate on the bottom in a heap just underneath the axis of the revolving magnet and around the bottom edge of the flask.
A much superior type of stirrer for microcarriers suspended in liquid culture medium is disclosed in U.K. Patent Specification 1485741. Such a stirrer is basically comprised of a stirring rod for immersion in liquid to be stirred, the stirring rod being provided with means for imparting to it an oscillatory pivotal motion such that the end of the rod describes an orbital path within the liquid being stirred.
With such a stirrer no accumulation occurs at the centre of the bottom of the flask in which the stirrer is mounted and the diameter of the tip orbit during its oscillatory movement can be made large enough to sweep around in the bottom edge of the flask.
However, it has been found that cells can be damaged in a large flat-bottomed flask because of the speed of rotation necessary to keep the microcarriers in suspension.
Objects ofthe invention An object of the invention is to provide a flask for use in growing anchorage-dependent cells in a stirred liquid culture medium containing microcarriers which is designed so that the speed of revolution of the stirrer can be low enough to avoid damage to the cells being swept around in the culture medium while at the same time maintaining the microcarriers in uniform suspension and obviating the risk of accumulation of the microcarriers at the centre or edge of the flask bottom.
Summary of the invention According to the invention therefore there is provided a flask for use in living cell growth techniques in a culture medium comprising a base formed of a conical portion which tapers to its apex in an upward direction from said base within the interior of said flask, said portion being delimited by an annular trough or channel forming the bottom corner of the flask, said trough or channel being radiused between the inner wall surface of the flask and the surface of said conical portion interior of the flask.
With such a configuration of the bottom of the flask the speed of revolution of the orbiting stirrer rod, if the flask is used with such a device, can be as low as 50 rpm or less without giving rise to accumulation of the microcarriers at the centre or edge of the flask bottom. Thus damage to the cells by the previously required high rotational or orbiting motion of the stirrer rod necessary to maintain the cells in uniform suspension, is eliminated, while at the same time accumulation of microcarriers at the bottom of the flask avoided.
Brief description of the drawings An embodiment of the invention will now be described by way of example with reference to the sole accompanying drawing which shows a crosssectional view of a flask according to the invention shown in its application for use with an oscillatory stirrer which can be mounted to the flask in some convenient manner.
Detailed description ofa preferred embodiment The flask shown in the sole accompanying drawing will be described as it may be employed with an oscillatory type stirring apparatus as described in the aforementioned United Kingdom Patent Specification 1485741. The stirring rod 1 of such apparatus itself since this is believed unnecessary to an understanding of the invention. Suffice it to say the the stirrer rod 1 of the stirring apparatus as mentioned can adopt several positions as shown such that it performs orbotal rotational movements, the stirring action closely resembling that of an ordinary hand-held stirring rod.
The strirrer rod 1 is shown mounted in a flask 2 for containing, when the flask is used in connection with the growth of living cells, microcarriers or beads on which anchorage-dependent cells can be grown in a liquid culture medium suspension.
The bottom of the flask 2 is formed of a conical portion 3 which tapers to its apex 4 in an upward direction to the interior of flask 2.
The conical portion 3 is delimited by an annular trough or channel 5. The channel or trough 5 is radiused as shown between the surface of the conical portion 3 facing interiorly of the flask 2 and the interior surface 6 of wall 7 of the flask 2.
The conical portion 3 is formed in the manner of a concave depression in the base of the flask 2 and is therefore of inverted v-shape in cross-section as shown.
The height of the apex of the cone-shaped portion 3 from the base of the cone is of the order of one third of the diameter D of the flask 2 and the diameter of the cone of the order of one half the diameter of the flask 2. Moreover, the radius of the annular trough or channel 5 is of the order of one eighth of the diameter D of the flask 2.
In the embodiment as described the axis of the cone-shaped portion 3 lies on the central vertical axis 8 of the flask 2.
In operation the stirrer rod 1 is arranged to sweep along the bottom of the flask as shown with an orbital motion so that it traces the annular trough or channel 5.
With the configuration of the bottom of the flask as described uniformity of microcarrier suspension is achieved at stirrer rod speeds much lower than those necessary to maintain the same uniformity in a flat-bottomed flask.
Although the flask of the invention has been described with reference to its use in living cell growth techniques, nevertheless it will be appreciated that it has wider applications for example in chemical mixing and the like.

Claims (6)

1. Aflaskfor use in living cell growth techniques in a culture medium comprising a base formed of a conical portion which tapers to its apex in an upward direction from said base within the interior of said flask, said portion being delimited by an annular trough or channel forming the bottom corner of the flask, said trough or channel being radiused between the inner wall surface of the flask and the surface of said conical portion interior of the flask.
2. A flask as claimed in Claim 1 wherein said conical portion is hollow, being a depression formed in said base.
3. A flask as claimed in Claim 2 wherein the height of said coned-shaped portion on the base of said cone is of the order of one third of the diameter of the flask.
4. Aflask as claimed in any preceding claim wherein the diameter of said cone-shaped portion is of the order one half the diameter of the flask.
5. A flask as claimed in any preceding claim wherein the radius of the annular trough or channel is of the order of one eighth of the diameter of the flask.
6. Aflaskfor use in living cell growth techniques in a culture medium substantially as hereinbefore described with reference to the drawing.
GB7924810A 1979-07-17 1979-07-17 A flask for use in stirring particles in suspension such as microcarriers for anchorage-dependent living cells in a liquid culture medium Withdrawn GB2053704A (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
GB7924810A GB2053704A (en) 1979-07-17 1979-07-17 A flask for use in stirring particles in suspension such as microcarriers for anchorage-dependent living cells in a liquid culture medium
GB8023215A GB2054397B (en) 1979-07-17 1980-07-16 Method and apparatus for stirring particles in suspension such as microcarriers for anchorage-dependent living cells in a liquid culture medium
US06/277,403 US4382685A (en) 1979-07-17 1981-06-25 Method and apparatus for stirring particles in suspension such as microcarriers for anchorage-dependent living cells in a liquid culture medium

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
GB7924810A GB2053704A (en) 1979-07-17 1979-07-17 A flask for use in stirring particles in suspension such as microcarriers for anchorage-dependent living cells in a liquid culture medium

Publications (1)

Publication Number Publication Date
GB2053704A true GB2053704A (en) 1981-02-11

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GB7924810A Withdrawn GB2053704A (en) 1979-07-17 1979-07-17 A flask for use in stirring particles in suspension such as microcarriers for anchorage-dependent living cells in a liquid culture medium
GB8023215A Expired GB2054397B (en) 1979-07-17 1980-07-16 Method and apparatus for stirring particles in suspension such as microcarriers for anchorage-dependent living cells in a liquid culture medium

Family Applications After (1)

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GB8023215A Expired GB2054397B (en) 1979-07-17 1980-07-16 Method and apparatus for stirring particles in suspension such as microcarriers for anchorage-dependent living cells in a liquid culture medium

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2183338A (en) * 1985-11-19 1987-06-03 Sclavo Spa Self-mixing cell for photometric and turbidimetric measurements
US5267791A (en) * 1991-12-13 1993-12-07 Corning Incorporated Suspended cell culture stirring vessel closure and apparatus
EP1516920A1 (en) * 2003-09-19 2005-03-23 The Automation Partnership Cell culture vessel for the automated processing of cell cultures
CN118126928A (en) * 2024-01-31 2024-06-04 广东永顺生物制药股份有限公司 Microcarrier culture method of adherent cells

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3165197D1 (en) * 1980-12-04 1984-09-06 Techne Cambridge Ltd Method and apparatus for stirring particles in suspension such as microcarriers for anchorage-dependent living cells in a liquid culture medium
US4498785A (en) * 1982-06-09 1985-02-12 Techne Corporation Floating magnetic stirrer for culture medium

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2183338A (en) * 1985-11-19 1987-06-03 Sclavo Spa Self-mixing cell for photometric and turbidimetric measurements
US5267791A (en) * 1991-12-13 1993-12-07 Corning Incorporated Suspended cell culture stirring vessel closure and apparatus
EP1516920A1 (en) * 2003-09-19 2005-03-23 The Automation Partnership Cell culture vessel for the automated processing of cell cultures
CN118126928A (en) * 2024-01-31 2024-06-04 广东永顺生物制药股份有限公司 Microcarrier culture method of adherent cells

Also Published As

Publication number Publication date
GB2054397B (en) 1983-03-02
GB2054397A (en) 1981-02-18

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