CN201746556U

CN201746556U - Internal recycle air lift type cell culture device

Info

Publication number: CN201746556U
Application number: CN 201020257596
Authority: CN
Inventors: 潘炘
Original assignee: Zhejiang Academy of Forestry
Current assignee: Zhejiang Academy of Forestry
Priority date: 2010-07-12
Filing date: 2010-07-12
Publication date: 2011-02-16
Anticipated expiration: 2020-07-12

Abstract

The utility model relates to an inner recycle air lift type cell culture device, which is mainly suitable for cell culture of agricultural and forestall tissues. The internal recycle air lift type cell culture device is characterized in that an air disperser, a culture flask aspirating hole and a culture flask air inlet hole are arranged on a cell culture flask; the air disperser and the culture flask air inlet hole are both positioned at the middle lower part of the flask body, and the culture flask aspirating hole is positioned at the middle upper part of the flask body; an interior air circulating mechanism is arranged and comprises a transmission permanent seat, a wear-resisting rope, a pair of piston bodies installed opposite to each other, a pair of offsetting springs and a pair of offsetting spring clips; the interior air circulating mechanism is connected with the cell culture flask; and piston air inlet holes and piston air outlet holes are arranged on the piston bodies. The utility model has reasonable structural design, and has the advantage that air in a cell reactor is circulated into culture solution through a small-size one-way piston pump so that cells of the plant tissues in the cell culture flask are in an air lift culture state. In addition, the device is easy to detach, assemble and generalize and has low cost, and the damage and replacement do not influence the operation.

Description

Internal circulation gas-lift type cell culture apparatus

Technical field

The utility model relates to a kind of cell culture apparatus, particularly a kind of internal circulation gas-lift type cell culture apparatus, it mainly is applicable to the cell cultures of agroforestry tissue, especially be fit to shearing force and to the external source gas sensitization to obtain the cultivation that important secondary metabolite is the medicinal plant cell of purpose.

Background technology

The device that is used for plant tissue cell's cultivation at present is a lot, and vegetable cell can synthesize many secondary metabolites with important value, comprises active medicinal matter.Utilize the extensive suspension culture vegetable cell of bio-reactor to produce the plant desirable metabolites and obtained very great development in recent years, DESIGN OF REACTOR and development aspect mainly focus on the design and the making of the bio-reactor that mixing efficiency is higher, shearing resistance is lower.

The reactor of at present suitable plant tissue and cell mainly contains the mechanical stir-reactor and the airlift reactor that utilizes gas injection kinetic energy by the mechanical stirring intake.

Mechanical agitation type be 20th century the seventies grow up, its main drawback is that the cell walls of vegetable cell is poor to the tolerance of shearing, the axle envelope between stirring type bioreactor stir shaft and tank body is often leaked easily and is caused microbiological contamination.

Gas is given birth to the formula bio-reactor and is relied on a large amount of ventilation entering momentums and energy, with good heat transfer, the mass transfer of nutrient solution in the assurance reactor, and guarantees not occur the dead angle.Because unique hydrodynamic characteristic, airlift reactor has following characteristics: caloic transfer rate height, and air feed efficient height, simple in structure, inner movement-less part, the air flow height, fluid effect is splendid, and waste of energy is even.

In the prior art, have lot of advantages, its shortcoming is also arranged for the little volume cell cultures in laboratory though gas is given birth to the formula bio-reactor; Because its source of the gas all is that air compressor machine or gas cylinder pump in the reactor by pressure difference, wherein need be through independent filtration unit to reach the sterilization purpose; Therefore, cause equipment complicated and expensive, energy consumption is also high; In addition, external source of the gas feeds has driven away carbonic acid gas and the ethene in the enchylema, and the growth of pair cell has inhibition on the contrary, and the oxygen uptake rate of vegetable cell is lower, and too high dissolved oxygen is unfavorable to the synthetic secondary metabolite of vegetable cell.

China's utility model patent numbers 902258052 discloses a kind of " high viscosity fermentation gas is given birth to the formula bio-reactor ", its be a kind of in circulation high viscosity fermentation gas life formula bio-reactor, can be used for microbial fermentation and cell culture of animals; This reactor structural disadvantages in actual use is: it mainly is to cultivate at the mycelia ferment, although its applicable culture plant cell still can injure responsive vegetable cell when managing the lifter motion process in this reactor.Its required air therefore also belong to external source gas lift culture plant cell and culture apparatus thereof, and this device external air source pressure is excessive, and is not suitable for similar ginkgo, the cultivation of yew cell from the outside.

Chinese invention patent application number 031499058 discloses a kind of " have the screen cloth type and lead the living formula vegetable cell of the gas reactor of irritating tube ", this reactor structural disadvantages in actual use is: need use air filter when it uses, and its source of the gas comes from outside air compressor machine, belongs to external source gas lift culture plant cell and culture apparatus thereof; Simultaneously, not air but come gas in the autoreactor, still need air filter so can't accomplish sterile state because of air compressor machine even its external source air compressor machine is employed.

Chinese invention patent application number 2004100007971 discloses a kind of " cycle submergence gas is given birth to the culture plant cell and the culture apparatus thereof of formula ", it mainly is applicable to the vegetable cell that needs control cell aggregation size and need immobilization and suspension culture, this culture apparatus structural disadvantages in actual use is: complex structural designs, need external gas holder, and the air filtration post, also the membrane type air filter has been used in many places in addition; Therefore it is a kind of external source formula gas lift culture plant cell and culture apparatus thereof, is not suitable for endogenous gas lift culture plant cell.

The utility model content

Technical problem to be solved in the utility model is to overcome the equipment complexity that existing use external air source causes in the prior art, and the ventilation rear-guard is except defectives such as carbonic acid gas in the enchylema and ethene, and a kind of reasonable in design is provided, can cultivates the internal circulation gas-lift type cell culture apparatus of medicinal plant cell.

The utility model solves the problems of the technologies described above the technical scheme that is adopted: this internal circulation gas-lift type cell culture apparatus comprises Tissue Culture Flask, is provided with air dispersion device with holes in the bottle, it is characterized in that:

Also have the culturing bottle aspirating hole on the described Tissue Culture Flask, the culturing bottle air inlet port; Described air dispersion device and culturing bottle air inlet port all are positioned at the middle and lower part of bottle, and the culturing bottle aspirating hole is positioned at the middle and upper part of bottle.

The lower surface of described air dispersion device communicates with the culturing bottle air inlet port.

Be provided with the internal gas cycling mechanism, comprise the transmission permanent seat, wear-resisting rope, a pair of piston body of installing in opposite directions, a pair of return spring and a pair of return spring card; This internal gas cycling mechanism links to each other with described Tissue Culture Flask.

Have piston intake hole and piston air outlet on the described pair of pistons body.

Described return spring contacts with piston body, and return spring fastens on the transmission permanent seat, and the position and the return spring of this return spring card are complementary, and described wear-resisting rope is connected with piston body.

As preferably, piston body described in the utility model comprises stopcock and stopcock seat, and stopcock is positioned at the stopcock seat, and can move in the stopcock seat; One end of described return spring is positioned at the stopcock seat and contacts with stopcock, and an end of described wear-resisting rope is connected with stopcock.

As preferably, be provided with also on the internal gas cycling mechanism described in the utility model that rolling disc, flake down encircle admittedly and a pair of rope set collar; Described rolling disc is between the pair of pistons body, and flake down encircles admittedly and is fixed on the rolling disc, and the rope set collar is fixed on the transmission permanent seat, and described wear-resisting rope passes the rope set collar and encircles admittedly with rolling disc with flake down and be connected.

As preferably, also be provided with power supply, motor and current regulator power supply on the internal gas cycling mechanism described in the utility model; Described motor is connected with rolling disc, and power supply then links to each other with motor respectively with current regulator.

As preferably, Tissue Culture Flask of the present utility model links to each other with the internal gas cycling mechanism by threeway.

As preferably, the culturing bottle aspirating hole of Tissue Culture Flask described in the utility model links to each other with the pair of pistons air inlet port of internal gas cycling mechanism by the glass threeway, and the culturing bottle air inlet port of Tissue Culture Flask links to each other with the pair of pistons production well of internal gas cycling mechanism by the glass threeway.

As preferably, the utility model also is provided with and has the gas external treatment bottle of handling the bottle air inlet port and handling the bottle production well, and processing bottle production well wherein links to each other with the culturing bottle air inlet port, handles bottle air inlet port and links to each other with piston air outlet on the piston body.

The utility model compared with prior art, have the following advantages and effect: reasonable in design, by the small unidirectional piston pump is that the internal gas cycling mechanism pumps into the gas circulation in the Tissue Culture Flask in the nutrient solution, solved existing following defective in the prior art: external source gas takes place with the filter apparatus configuration design complicated, and external air source ventilation rear-guard is except carbonic acid gas in the enchylema and ethene, and too high dissolved oxygen is unfavorable to the synthetic secondary metabolite of vegetable cell.The used air cycle of the utility model mechanism can dismantle sterilization, and do not have when using microbiological contamination may, therefore, need not external gas-filtering device, promptly saved cost, also simplified system architecture.Certainly, also can reach and improve endogenous gaseous fraction, replenish the essential oxygen composition of plant by external gas processing bottle.

Simultaneously, this device in use, a pair of stopcock pumps the intravital air of Tissue Culture Flask respectively incessantly with motor and rolling disc, makes the cell of cell cultures bottle implants tissue be in the gas lift cultivation conditions; In addition, because adjustable economize on electricity engine power reaches the function of controlling air flow, to reach best gas lift culture effect; Whole device can be taked modular design, and the convenient inventiona gas treatment module that inserts is to strengthen whole device usefulness; Whole apparatus structure simplicity of design, dismounting and assembling easily; Whole device promptly can whole be sterilized, the back assembling use again behind the inoculation histocyte in the aseptic technique platform of also can sterilizing respectively; Each parts is taked modular design, and each assembly damages and changes the operation that does not influence whole device, so production cost is low, is easy to apply.

Description of drawings

Fig. 1 is the structural representation of the utility model diplopore culturing bottle;

Fig. 2 is the structural representation front view of the utility model internal gas cycling mechanism;

Fig. 3 is the structural representation vertical view of Fig. 2;

Fig. 4 is an one-piece construction synoptic diagram of the present utility model;

Fig. 5 is the structure enlarged diagram at A place among Fig. 3;

Fig. 6 is the structural representation of gas external treatment bottle of the present utility model.

Among the figure: 1 air dispersion device, 2 bottles, 3 cultivate bottle cap, 4 culturing bottle aspirating holes, 5 culturing bottle air inlet ports, 6 piston air outlet, 7 piston intake holes, 8 stopcocks, 9 stopcock seats, 10 return spring cards, 11 rolling discs, 12 return springs, 13 rope set collars, 14 flake downs encircle admittedly, 15 wear-resisting ropes, 16 transmission permanent seats, 17 power supplys, 18 motors, 19 current regulators, 20 handle the bottle production well, and 21 handle bottle air inlet port, 22 gas external treatment bottles, 23 glass threeways, 24 flexible pipes, I Tissue Culture Flask, II internal gas cycling mechanism, II-1 piston body, II-11 left piston body, the right piston body of II-12, II-13 left piston body bottom, the right piston body of II-14 top.

Embodiment

Below in conjunction with accompanying drawing embodiment of the present utility model is elaborated, present embodiment provides detailed embodiment and concrete operating process, but protection domain of the present utility model is not limited to following embodiment.

Referring to Fig. 1-Fig. 6, present embodiment mainly is made up of Tissue Culture Flask I and internal gas cycling mechanism II two portions.

The Tissue Culture Flask I of present embodiment is provided with air dispersion device 1 with holes, culturing bottle aspirating hole 4, culturing bottle air inlet port 5; Air dispersion device 1 and culturing bottle air inlet port 5 all are positioned at the middle and lower part of bottle 2, and culturing bottle aspirating hole 4 is positioned at the middle and upper part of bottle 2; The lower surface of described air dispersion device 1 communicates with culturing bottle air inlet port 5.

The internal gas cycling mechanism II of present embodiment, it comprises transmission permanent seat 16, wear-resisting rope 15, a pair of piston body II-1 that installs in opposite directions, a pair of return spring 12 and a pair of return spring card 10; This internal gas cycling mechanism II links to each other with Tissue Culture Flask I; Have piston intake hole 7 and piston air outlet 6 on the pair of pistons body II-1; Return spring 12 contacts with piston body II-1, and return spring card 10 is fixed on the transmission permanent seat 16, and the position of this return spring card 10 and return spring 12 are complementary, and wear-resisting rope 15 is connected with piston body II-1.

The said a pair of piston body II-1 that installs in opposite directions is meant in the literary composition: be positioned at the left end of left piston body II-11 into and out of pore on the left piston body II-11, be positioned at the right-hand member of right piston body II-11, the symmetrical configuration of left and right piston body II-11, II-12 into and out of pore on the right piston body II-12.

Described piston body II-1 comprises stopcock 8 and stopcock seat 9, and stopcock 8 is positioned at stopcock seat 9, and can move in stopcock seat 9; One end of described return spring 12 is positioned at stopcock seat 9 and contacts with stopcock 8, and an end of described wear-resisting rope 15 is connected with stopcock 8.

Also be provided with rolling disc 11, flake down on the described internal gas cycling mechanism II and encircle 14 and a pair of rope set collar 13 admittedly; Rolling disc 11 is between pair of pistons body II-1, and flake down encircles 14 admittedly and is fixed on the rolling disc 11, and rope set collar 13 is fixed on the transmission permanent seat 16, and wear-resisting rope 15 passes rope set collar 13 and flake down and encircles 14 admittedly and be connected with rolling disc 11.

Also be provided with power supply 17, motor 18 and current regulator power supply 19 on the described internal gas cycling mechanism II; Motor 18 is connected with rolling disc 11, and power supply 17 links to each other with motor 18 respectively with current regulator 19.

The culturing bottle aspirating hole 4 of described Tissue Culture Flask I links to each other with the pair of pistons air inlet port 7 of internal gas cycling mechanism II by glass threeway 23, and the culturing bottle air inlet port 5 of Tissue Culture Flask I links to each other with the pair of pistons production well 6 of internal gas cycling mechanism II by glass threeway 30.

As option, present embodiment also can be provided with and have the gas external treatment bottle 22 of handling bottle air inlet port 21 and handling bottle production well 20, processing bottle production well 20 wherein links to each other with culturing bottle air inlet port 5, handles bottle air inlet port 21 and links to each other with piston air outlet 6 on the piston body II-1.

In the present embodiment, piston body II-1 is installed in the transmission permanent seat 16 by bayonet socket, and an end of stopcock 8 is equipped with return spring 12 and wear-resisting rope 15, and the other end of wear-resisting rope 15 links to each other with rolling disc 11.When mounted, should guarantee a bottom that as far as possible is in piston body II-1 in a pair of stopcock 8, as the bottom II-13 of left piston body II-11, another is in the top of piston body II-1 as far as possible, as the top II-14 of right piston body II-12.

Tissue Culture Flask I can be triangular flask, and bottle 2 volumes can be 250ml, 500ml, 1000ml or bigger.

Piston body II-1 and bottle link to each other by high temperature resistant PE flexible pipe 24 or primary sterilization PV flexible pipe and glass threeway 23.High temperature resistant PE flexible pipe 24, and piston body II-1 and bottle 2 etc. all can place 121 ℃ of autoclavings to handle.

The adjustable power of described rolling disc 11 is with the adjustments of gas circulation velocity.

In addition, the flexible pipe 24 that is connected with the culturing bottle air inlet port 5 of Tissue Culture Flask I can insert inventiona gas treatment module by gas folder and glass threeway 23, and this module can increase arbitrarily, with the air in the daf molecule culturing bottle I or increase a certain or several particular components.

In the present embodiment: cultivate bottle cap 3 and be the high-temperature resistance plastice lid, power supply 17 is the 12V power supply, and stopcock seat 9 is a demountable structure, and the hole on the air dispersion device 1 is small through hole.

During use,, the gas among the Tissue Culture Flask I is extracted out by culturing bottle aspirating hole 4, and thrown into, in gas distributor 1 homodisperse liquid nutrient medium thereon by culturing bottle air inlet port 5 by internal gas cycling mechanism II.

The principle of work of internal gas cycling mechanism II is as follows: motor 18 rotates rolling disc 11 under power supply 17 drives, and then encircles the wear-resisting rope 15 of 14 pullings admittedly by flake down, makes stopcock 8 reciprocating under return spring 12 and 15 effects of wear-resisting rope.Because transmission permanent seat 16 by a pair of stopcock seat 9 is housed, therefore forms the continuity of bleeding and giving vent to anger.Wherein return spring 12 is positioned stopcock seat 9 inside by return spring card 10.In addition, by the power of current regulator 19 may command motors 18, and then the travelling speed of control rolling disc 11, thereby reach the purpose of controlling airshed.

Gas external treatment bottle 22 is a selectable unit (SU), if need to add these parts, only needs by high temperature resistant PE flexible pipe 24 or disposable sterilization plastics tubing these parts are connected with the utility model and gets final product, to reach purpose such as oxygenation.

The utility model can bulk-breaking, also can connect part becomes integral body to carry out vapor sterilization, after in Tissue Culture Flask I, injecting the certain amount of fluid substratum, and can in the aseptic technique platform, inoculate plant tissue cell, connect the effect that power supply 17 backs just can reach the air lift type cell cultures through sterilization.

The Tissue Culture Flask I cumulative volume of this enforcement is 500ml, and the air input that piston body II-1 makes a round trip is 2ml.Present embodiment is a multi-functional culture plant cell device, can be used for medicinal plant tissue and the cultivation of cell short run in the laboratory.

Above content described in this specification sheets only is to the explanation of the utility model structure example; And the title that the utility model component are got also can be different, allly conceive equivalence or the simple change that described structure, feature and principle are done according to the utility model patent, are included in the protection domain of the utility model patent.

Claims

1. an internal circulation gas-lift type cell culture apparatus comprises Tissue Culture Flask, is provided with air dispersion device with holes in the bottle, it is characterized in that:

(1) also has the culturing bottle aspirating hole on the described Tissue Culture Flask, the culturing bottle air inlet port; Described air dispersion device and culturing bottle air inlet port all are positioned at the middle and lower part of bottle, and the culturing bottle aspirating hole is positioned at the middle and upper part of bottle;

(2) lower surface of described air dispersion device communicates with the culturing bottle air inlet port;

(3) be provided with the internal gas cycling mechanism, it comprises the transmission permanent seat, wear-resisting rope, a pair of piston body of installing in opposite directions, a pair of return spring and a pair of return spring card; This internal gas cycling mechanism links to each other with described Tissue Culture Flask;

(4) have piston intake hole and piston air outlet on the described pair of pistons body;

(5) described return spring contacts with piston body, and return spring fastens on the transmission permanent seat, and the position and the return spring of this return spring card are complementary, and described wear-resisting rope is connected with piston body.

2. internal circulation gas-lift type cell culture apparatus according to claim 1 is characterized in that: described piston body comprises stopcock and stopcock seat, and stopcock is positioned at the stopcock seat; One end of described return spring is positioned at the stopcock seat and contacts with stopcock, and an end of described wear-resisting rope is connected with stopcock.

3. internal circulation gas-lift type cell culture apparatus according to claim 1 and 2 is characterized in that: be provided with also on the described internal gas cycling mechanism that rolling disc, flake down encircle admittedly and a pair of rope set collar; Described rolling disc is between the pair of pistons body, and flake down encircles admittedly and is fixed on the rolling disc, and the rope set collar is fixed on the transmission permanent seat, and described wear-resisting rope passes the rope set collar and encircles admittedly with rolling disc with flake down and be connected.

4. internal circulation gas-lift type cell culture apparatus according to claim 3 is characterized in that: also be provided with power supply, motor and current regulator power supply on the described internal gas cycling mechanism; Described motor is connected with rolling disc, and power supply then links to each other with motor respectively with current regulator.

5. internal circulation gas-lift type cell culture apparatus according to claim 1 and 2 is characterized in that: described Tissue Culture Flask links to each other with the internal gas cycling mechanism by threeway.

6. internal circulation gas-lift type cell culture apparatus according to claim 5, it is characterized in that: the culturing bottle aspirating hole of described Tissue Culture Flask links to each other with the pair of pistons air inlet port of internal gas cycling mechanism by the glass threeway, and the culturing bottle air inlet port of Tissue Culture Flask links to each other with the pair of pistons production well of internal gas cycling mechanism by the glass threeway.

7. internal circulation gas-lift type cell culture apparatus according to claim 1, it is characterized in that: also be provided with and have the gas external treatment bottle of handling the bottle air inlet port and handling the bottle production well, processing bottle production well wherein links to each other with the culturing bottle air inlet port, handles the bottle air inlet port and links to each other with piston air outlet on the piston body.