CN101974424A - Swing-driven circular cell culture device - Google Patents
Swing-driven circular cell culture device Download PDFInfo
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- CN101974424A CN101974424A CN2010105329125A CN201010532912A CN101974424A CN 101974424 A CN101974424 A CN 101974424A CN 2010105329125 A CN2010105329125 A CN 2010105329125A CN 201010532912 A CN201010532912 A CN 201010532912A CN 101974424 A CN101974424 A CN 101974424A
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- liquid storage
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- 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
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- 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
- C12M27/24—Draft tube
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Abstract
The invention relates to a swing-driven circular cell culture device. The culture device comprises a hard culture container the lower part of which is provided with a solution chamber; the bottom surface of the solution chamber is a smooth concave surface; a slope surface is arranged on the side wall surface of the solution chamber; the slope surface and the concave surface are in smooth transition; a circular pipe is arranged on the side wall of the solution chamber along the slope surface in a matched manner; the circular pipe is provided with an upper opening and a lower opening; and a one-way discharge valve structure is arranged at the lower opening. When the circular cell culture device is driven to swing left and right, the upper culture solution constantly enters from the upper openings of the two circular pipes and rushes out from the lower openings, while the original lower culture solution is turned upwards in the process to realize up-down circulation of the culture solution; the circulation is simple and effective, and the mixing and ventilation effect is good; and moreover, the circulation frequency can be minimized and the shearing force of the circulation is little.
Description
Technical field
The present invention relates to a kind ofly wave when driving applying for it, the nutrient solution in it can carry out the round-robin cell culture apparatus.
Background technology
The cultivation of organism (comprises zooblast, vegetable cell, the cultivation of insect cell and microorganism) and utilize the organism of above-mentioned cultivation to carry out biological technology products (as protein, the polypeptide class, the nucleic acid class, antibody class, virus vector and virus vaccines class) production, need provide suitable temperature, necessary gas (as oxygen and carbon dioxide) and suitable mixing, therefore the various products that adapt have with it appearred on the market, as thermostat container, CO2gas incubator is mixed shaking table, fermentor tank, culturing bottle and various cell biological reactors etc.When carrying out cell cultures, animal cell culture and when utilizing zooblast to carry out the production of above-mentioned biological technology products particularly, often need precise dose control, good gas and nutrition exchange and the very little mixing of shearing force, therefore on the market various bio-reactors have appearred, as roll a bottle bio-reactor, magnetic agitation bio-reactor, basket bio-reactor of NBS and WAVE wave bio-reactor etc.
The containers for culturing organisms that is used for animal cell culture in the market mainly contains rolling bottle, the magnetic agitation culturing bottle, basket resting cell culture bioreactors of NBS and WAVE shake the pocket type bio-reactor, wherein rolling bottle and magnetic agitation culturing bottle only are used for small-scale production and amplify step by step, because the easy crossed contamination of its non-once use.In addition, magnetic agitation is cultivated and is existed the bigger shortcoming of shearing force.The basket resting cell culture bioreactors of NBS costs an arm and a leg, and complicated operation is unsuitable for the laboratory and cultivates on a small scale, and there is cross-contamination issue in its non-once culture tank.It is that cell cultures is cultivated in the soft bag of disposable plastic that WAVE shakes the pocket type bio-reactor, produces wave by shaking bag, makes cell suspension, nutrient solution mixes, gaseous interchange, its shearing force is less relatively, but has following shortcoming: at first, WAVE mainly is applicable to large-scale cell cultures, tool WAVE patent shows that volume of culture is preferably in more than the 5L, when small volume is cultivated, can not produce enough waves and make cell suspension, cell deposition at the bottom of bag, phase mutual friction in the process of shaking and damaging; Secondly, it is the soft bags of plastics that WAVE shakes bag, adds continuous a series of pipelines, and operation is than hardware culture vessel complexity; Once more, the WAVE culture bag of different model will be with the rocker that matches, and pouch portion can use big rocker, and sack can not be used little rocker, need buy the equipment of a series of costlinesses through being amplified to large-scale industrial production step by step from cultivating on a small scale; At last,, produce necessary cell suspension, mix fully and gaseous interchange, also want the certain speed of waving and frequency, therefore also can produce shearing force to a certain degree though WAVE is little than the stirring type bioreactor shearing force.
In order to overcome the shortcoming of above-mentioned bio-reactor, particularly the deficiency of WAVE swing reactor must be researched and developed novel cell culture apparatus.
Summary of the invention
The object of the present invention is to provide a kind of shearing force little, mix the good circulating type cell incubator that waves driving of ventilation effect.
For achieving the above object, the present invention adopts following technical scheme: a kind of circulating type cell incubator that waves driving, this culture apparatus comprises the hard culture vessel, the bottom of this incubator is a liquid storage chamber, and the bottom surface of liquid storage chamber is slick concave surface, and the side wall surface of liquid storage chamber is a slope, seamlessly transit between slope and the concave surface, be provided with circulating line along slope is identical on the sidewall of liquid storage chamber, circulating line has upper shed and lower opening, and lower opening is provided with unidirectional venting valve structure.
Described circulating line is arranged on film on the side wall surface of liquid storage chamber and liquid storage chamber by both side edges sidewall bag is established and is formed.
Described circulating line is formed by the body on the sidewall that is installed in liquid storage chamber that coincide.
Described unidirectional venting valve structure is plural bag shape flexible valve, and the sack of bag shape flexible valve is towards the port of lower opening, and the outer side correspondence of bag shape flexible valve is arranged on the inwall of circulating line.
Described circulating line is a soft pipeline, and bag shape flexible valve and circulating line are wholely set.
Described unidirectional venting valve structure is the paddle type limit diaphragm that coincide with the pipe port shape of body, and the limit diaphragm folding is sticked on the lower opening of body, and the outer rim of limit diaphragm is less than the periphery of body lower opening.
The residing plane, edge of residing plane, the edge of the upper shed of described circulating line and lower opening is to the inclined-plane of the bottom surface of liquid storage chamber inclination, and the upper shed of circulating line is provided with the drainage buffering mantle edge that extends to the upper shed center of circulating line.
Described liquid storage chamber is the semielliptical shape, and the inwall of liquid storage chamber is oval arcuation, and described circulating line is two, and two circulating lines are symmetricly set on the both ends of ellipsoid major diameter respectively.
The one that makes progress on the opening on described liquid storage chamber top extension is provided with column and holds air chamber, and the upper end open of holding air chamber is provided with container cover, and the sidewall that holds air chamber is provided with ventilative window, and ventilative window is provided with the degerming filter membrane.
The one that makes progress on the opening on described liquid storage chamber top extension is provided with column and holds air chamber, hold that sealing is provided with container cover on the upper end open of air chamber, hold on the sidewall of air chamber or be communicated with at interval on the container cover be provided with the air inlet mouth of pipe, the mouth of pipe of giving vent to anger, the liquid in-out mouth of pipe, sampling nozzle.
The bottom surface of liquid storage chamber of the present invention is slick concave surface, the side wall surface of liquid storage chamber is a slope, seamlessly transit between slope and the concave surface, on the sidewall of liquid storage chamber, coincide and be provided with circulating line along slope, when driving the present invention vacillates now to the left, now to the right, being arranged in the nutrient solution on upper strata can be constantly be entered circulating line and is gushed out by lower opening by the upper shed of two circulating lines, and the nutrient solution that originally is in lower floor turns on can be in this process, realize the circulation up and down of nutrient solution, simultaneously, because the lower opening of circulating line is provided with unidirectional venting valve structure, the liquid that enters circulating line only may be supernatant liquid, and circulation is effectively simple, and it is good to mix ventilation effect, and the round-robin frequency can drop to Schwellenwert, and circulation shear power is minimum.Simultaneously, culture vessel of the present invention is the hard culture vessel, moves conveniently.
Circulating line of the present invention is formed by the body on the sidewall that is installed in liquid storage chamber that coincide, and body can separate moulding with culture vessel, and manufacturing process is better.
Unidirectional venting valve structure of the present invention is plural bag shape flexible valve, the sack of bag shape flexible valve is towards the port of lower opening, banded limit diaphragm is arranged on the inwall of circulating line by a side, wave in driving the present invention like this, when the upper shed position of circulating line reduces, nutrient solution can enter in the circulating line from upper shed, the nutrient solution of lower opening will be full of a bag shape flexible valve, bag shape flexible valve expands lower opening is shut, when the upper shed position of circulating line raises, nutrient solution squeeze bag shape flexible valve in the circulating line, liquid in the bag shape flexible valve is extruded, bag shape flexible valve smaller volume, and nutrient solution will be discharged by lower opening, this valve is a biomimetic features, unidirectional through-flow reliability height.
The one that makes progress on the opening on culture vessel of the present invention top extension is provided with column and holds air chamber, the upper end open of holding air chamber is provided with container cover, be communicated with at interval on the sidewall of appearance air chamber and be provided with inlet mouth and air outlet, when the volume of culture vessel is big, nutrient solution only contacts with air by culture vessel upper end open place and dissolved oxygen can't satisfy the demand the time, can be by inlet mouth oxygenation in the culture vessel, discharge the waste gas that organism produces by the air outlet simultaneously, can well guarantee the vegetative activity of organism.
Culture vessel of the present invention is the semielliptical shape, and inner chamber is oval arcuation, circulating line arrangement convenience like this, and circulation fluid moves on the long axis direction of oval ball back and forth gently, shearing force can be dropped to minimum.
Description of drawings
Fig. 1 is the stereographic map of the embodiment of the invention 1;
Fig. 2 is the sectional view of the front view of the embodiment of the invention 1;
Fig. 3 is the vertical view of Fig. 2;
Fig. 4 is the sectional view of the front view of the embodiment of the invention 2;
Fig. 5 is the vertical view of Fig. 4;
Fig. 6 is the perspective view of Fig. 5 middle tube body;
Fig. 7 is the stereographic map of the embodiment of the invention 3;
Fig. 8 is the front view of the embodiment of the invention 3;
Fig. 9 is the perspective view of Fig. 7 middle tube body;
Figure 10 is the stereographic map of bag shape flexible valve among Fig. 7.
Embodiment
A kind of embodiment 1 that waves the circulating type cell incubator of driving, in Fig. 1 ~ 3, this culture apparatus comprises the hard culture vessel, the top of this culture vessel is for holding air chamber 3, the bottom is a liquid storage chamber 4, and liquid storage chamber 4 is a semielliptical shape, the inner chamber ellipsoid cambered surface of liquid storage chamber 4, hold air chamber 3 and be wholely set, hold on the upper port of air chamber 3 by liquid storage chamber 4 extending upward with liquid storage chamber 4.The both sides that are relatively set with two films, 6, two films 6 at the both ends of the major axis of liquid storage chamber's elliposoidal inner chamber fixedly are sticked on the wall of inner chamber chamber.Culture vessel in the present embodiment is by the rigid plastics moulding, and film is a soft plastics, on the chamber wall of the inner chamber that film can be by hot melting mode fixed culture vessel.Two films 6 have been set as two body chambeies with the inner chamber of liquid storage chamber near the part bag at major axis two ends with the chamber wall of the inner chamber of liquid storage chamber as the part of circulating line, the upper end of circulating line has upper shed 7 up, and the lower end of circulating line has the lower opening 9 on the bottom surface that is positioned at liquid storage chamber.The inboard of lower opening 9 is provided with marsupial shape flexible valve 11, be provided with two altogether, the sack of bag shape flexible valve 11 is towards the port of lower opening 9, the bag top is towards the inside of torus, one side of a limit diaphragm fixedly installs on the inner side-wall of film, and a side of another end limit diaphragm is fixedly installed on the chamber wall of inner chamber.Bag shape flexible valve can be semi-moon shaped also can be the semielliptical shape.
Be equipped with container cover 14 on the upper end open of appearance air chamber 3, be oppositely arranged two ventilative windows of ventilation mutually on the sidewall of appearance air chamber, ventilative window is provided with degerming filter membrane 15.
The bottom of culture vessel is provided with base 5.
A kind of embodiment 2 that waves the circulating type cell incubator of driving, at Fig. 4, Fig. 5, among Fig. 6, the difference of present embodiment and embodiment 1 is: the circulating line of present embodiment is formed by body 10, body 10 coincide and is sticked on the sidewall of liquid storage chamber, the upper port of body 10 is upper shed 7, the lower port of body 10 is a lower opening 9, the upper surface of body 10 and lower surface are the inclined-plane to the inclination of the bottom surface of liquid storage chamber 4, therefore upper shed 7 and lower opening 9 are oval-shaped mouthful, limit diaphragm 8 is the oval lamellar body that coincide with the pipe port shape of body, limit diaphragm 8 be sticked the lower surface on, limit diaphragm 8 is fixed on the body at one end, can be with respect to the body open and close movement.The size of limit diaphragm 8 is greater than the internal diameter of body lower opening and less than the external diameter of body lower opening, and limit diaphragm 8 just blocks the mouth of pipe of body and the end face of whole body all can not covered.The edge of the lower opening of body be enlarging to male cone (strobilus masculinus), the edge of limit diaphragm 8 is the conical surface that the edge with the lower opening of body coincide.Body is a flexible duct, and upper shed place of body is provided with drainage buffering mantle edge 12.Drainage buffering mantle edge 12 is arranged on the upper surface of body, and extends to the mouth of pipe.
A kind of embodiment 3 that waves the circulating type cell incubator of driving, in Fig. 7 ~ 10, the difference of present embodiment and embodiment 2 is: the inboard of the lower opening 9 of present embodiment is provided with marsupial shape flexible valve 11, be provided with two altogether, the sack of bag shape flexible valve 11 is towards the port of lower opening 9, the bag top is towards the inside of torus, and two packed limit diaphragms are wholely set on the inner side-wall of body near the outside of tube wall.Bag shape flexible valve can be semi-moon shaped also can be the semielliptical shape.
Extend upward the appearance air chamber 3 that is provided with column on the opening on culture vessel top, can add container cover 14 on the upper end open of appearance air chamber 3, be symmetrically arranged with inlet mouth 2 air outlets 1 on the sidewall of appearance air chamber 3, inlet mouth 2 air outlets 1, all be communicated with appearance air chamber 3, also be provided with liquid in-out mouth, thief hole on the sidewall of appearance air chamber, the also pre-buried signal wire that is equipped with for dissolved oxygen sensor and pH value transmitter on the sidewall of appearance air chamber.In use also can on inlet mouth 2, air outlet 3, the degerming filter membrane be set, to prevent bacterial contamination.
The material of the culture vessel of above embodiment is duroplasts, also can be pottery or other mechanically resistant materials.
The bag shape flexible valve of above embodiment is two, also can be for more.
Above embodiment is provided with the appearance air chamber of column, and the appearance air chamber of column also can be set, and simultaneously container cover is not set yet, allows the air on culture vessel top that the microorganism needed oxygen of surviving is provided.
The inner chamber of the liquid storage chamber of present embodiment is an ellipsoid, and its inner chamber can be designed to by the bottom surface in actual use is slick concave surface, and the sidewall of liquid storage chamber is a slope, the Any shape that seamlessly transits between slope and the concave surface.
The circulation body of present embodiment can also be one-body molded with liquid storage chamber.
The circulating type cell incubator that waves driving among the above embodiment needs to be placed on the drive unit that produces oscillating motion in use, oscillating motion can make the cyclic motion of nutrient solution generation shown in Fig. 2,4,8 in the culture vessel, to reach the purpose that allows liquid circulation take a breath,, this oscillating motion makes nutrient solution round-robin purpose even if also reaching extremely gently.
The cell culture apparatus of above embodiment has under the situation of shearing force minimum, reaches maximum mixing and gaseous interchange effect, and is easy and simple to handle, with low cost, can disposablely use no crossed contamination.
In use, the cell culture apparatus of above embodiment is placed on swing incubator or other rocking equipments, the cell culture apparatuses of different sizes do not need the swing incubator or other rocking equipments that are complementary with it, same swing incubator or other rocking equipments can finish from laboratory scale to cultivate the demand of amplifying step by step again to large scale culturing.
Claims (10)
1. circulating type cell incubator that waves driving, it is characterized in that: this culture apparatus comprises the hard culture vessel, the bottom of this incubator is a liquid storage chamber, the bottom surface of liquid storage chamber is slick concave surface, the side wall surface of liquid storage chamber is a slope, seamlessly transits between slope and the concave surface, and coincideing along slope on the sidewall of liquid storage chamber is provided with circulating line, circulating line has upper shed and lower opening, and lower opening is provided with unidirectional venting valve structure.
2. the circulating type cell incubator that waves driving according to claim 1 is characterized in that: described circulating line is arranged on film on the side wall surface of liquid storage chamber and liquid storage chamber by both side edges sidewall bag is established and is formed.
3. the circulating type cell incubator that waves driving according to claim 1 is characterized in that: described circulating line is formed by the body on the sidewall that is installed in liquid storage chamber that coincide.
4. according to claim 1 or the 2 or 3 described circulating type cell incubators that wave driving, it is characterized in that: described unidirectional venting valve structure is plural bag shape flexible valve, the sack of bag shape flexible valve is towards the port of lower opening, and the outer side correspondence of bag shape flexible valve is arranged on the inwall of circulating line.
5. the circulating type cell incubator that waves driving according to claim 4 is characterized in that: described circulating line is a soft pipeline, and bag shape flexible valve and circulating line are wholely set.
6. the circulating type cell incubator that waves driving according to claim 3 is characterized in that:
Described unidirectional venting valve structure is the paddle type limit diaphragm that coincide with the pipe port shape of body, and the limit diaphragm folding is sticked on the lower opening of body, and the outer rim of limit diaphragm is less than the periphery of body lower opening.
7. the circulating type cell incubator that waves driving according to claim 1, it is characterized in that: the residing plane, edge of residing plane, the edge of the upper shed of described circulating line and lower opening is to the inclined-plane of the bottom surface of liquid storage chamber inclination, and the upper shed of circulating line is provided with the drainage buffering mantle edge that extends to the upper shed center of circulating line.
8. the circulating type cell incubator that waves driving according to claim 1, it is characterized in that: described liquid storage chamber is the semielliptical shape, the inwall of liquid storage chamber is oval arcuation, and described circulating line is two, and two circulating lines are symmetricly set on the both ends of ellipsoid major diameter respectively.
9. the circulating type cell incubator that waves driving according to claim 1, it is characterized in that: the one that makes progress on the opening on described liquid storage chamber top extension is provided with column and holds air chamber, the upper end open of holding air chamber is provided with container cover, the sidewall that holds air chamber is provided with ventilative window, and ventilative window is provided with the degerming filter membrane.
10. the circulating type cell incubator that waves driving according to claim 1, it is characterized in that: the one that makes progress on the opening on described liquid storage chamber top extension is provided with column and holds air chamber, hold that sealing is provided with container cover on the upper end open of air chamber, hold on the sidewall of air chamber or be communicated with at interval on the container cover be provided with the air inlet mouth of pipe, the mouth of pipe of giving vent to anger, the liquid in-out mouth of pipe, sampling nozzle.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010532912.5A CN101974424B (en) | 2010-11-05 | 2010-11-05 | Swing-driven circular cell culture device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010532912.5A CN101974424B (en) | 2010-11-05 | 2010-11-05 | Swing-driven circular cell culture device |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101974424A true CN101974424A (en) | 2011-02-16 |
| CN101974424B CN101974424B (en) | 2015-03-25 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201010532912.5A Expired - Fee Related CN101974424B (en) | 2010-11-05 | 2010-11-05 | Swing-driven circular cell culture device |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102618441A (en) * | 2012-03-14 | 2012-08-01 | 江苏农林职业技术学院 | Swinging type liquid intermittent submerged culture apparatus |
| CN103415610A (en) * | 2011-03-18 | 2013-11-27 | 通用电气健康护理生物科学股份公司 | Flexible bag for cultivation of cells |
| CN103911285A (en) * | 2014-03-08 | 2014-07-09 | 郑州威瑞生物技术有限公司 | Disposable bioreactor |
| CN109289958A (en) * | 2017-07-24 | 2019-02-01 | 郑州威瑞生物技术有限公司 | Clamshell pipe cap and clamshell test tube component |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN201545840U (en) * | 2009-09-21 | 2010-08-11 | 惠识瑶 | Packed bed cell bioreactor of stirring-free device |
| CN101812405A (en) * | 2010-04-30 | 2010-08-25 | 惠倪 | Polygonal plastic bag swing cell reactor and method for culturing animal cells |
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2010
- 2010-11-05 CN CN201010532912.5A patent/CN101974424B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN201545840U (en) * | 2009-09-21 | 2010-08-11 | 惠识瑶 | Packed bed cell bioreactor of stirring-free device |
| CN101812405A (en) * | 2010-04-30 | 2010-08-25 | 惠倪 | Polygonal plastic bag swing cell reactor and method for culturing animal cells |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103415610A (en) * | 2011-03-18 | 2013-11-27 | 通用电气健康护理生物科学股份公司 | Flexible bag for cultivation of cells |
| CN103415610B (en) * | 2011-03-18 | 2016-10-19 | 通用电气健康护理生物科学股份公司 | For cultivating the flexible bag of cell |
| CN102618441A (en) * | 2012-03-14 | 2012-08-01 | 江苏农林职业技术学院 | Swinging type liquid intermittent submerged culture apparatus |
| CN103911285A (en) * | 2014-03-08 | 2014-07-09 | 郑州威瑞生物技术有限公司 | Disposable bioreactor |
| CN109289958A (en) * | 2017-07-24 | 2019-02-01 | 郑州威瑞生物技术有限公司 | Clamshell pipe cap and clamshell test tube component |
| CN109289958B (en) * | 2017-07-24 | 2023-10-31 | 郑州威瑞生物技术有限公司 | Flip formula cap and flip formula test tube assembly |
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| CN101974424B (en) | 2015-03-25 |
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