CN102382766B - Tidal bioreactor for animal cell culture - Google Patents
Tidal bioreactor for animal cell culture Download PDFInfo
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- CN102382766B CN102382766B CN 201110387699 CN201110387699A CN102382766B CN 102382766 B CN102382766 B CN 102382766B CN 201110387699 CN201110387699 CN 201110387699 CN 201110387699 A CN201110387699 A CN 201110387699A CN 102382766 B CN102382766 B CN 102382766B
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- 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
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- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
- C12M41/18—Heat exchange systems, e.g. heat jackets or outer envelopes
- C12M41/22—Heat exchange systems, e.g. heat jackets or outer envelopes in contact with the bioreactor walls
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Abstract
The invention provides a tidal bioreactor for animal cell culture. The tidal bioreactor for the animal cell culture mainly comprises a culture medium accommodating tank (3) and a three-gas water jacket constant-temperature culture box (18), wherein a cell culture reaction tank (17) is arranged inside the three-gas water jacket constant-temperature culture box (18); the culture medium accommodating tank (3) is arranged inside a heating constant-temperature water tank (2), or a water interlayer outside the culture medium accommodating tank (3) is communicated with the heating constant-temperature water tank (2) through a hot water circulating pump (37); the cell culture reaction tank (17) is communicated with the culture medium accommodating tank (3) through a third peristaltic pump (16); the culture medium accommodating tank (3) is communicated with a culture medium supplementing or recycling bottle (30) through a second peristaltic pump (15); and the culture medium supplementing or recycling bottle (30) is communicated with the culture medium accommodating tank (3) through a first peristaltic pump (14). The tidal bioreactor for the animal cell culture is small in investment, small in occupied space, low in production cost and running cost, and high in yield.
Description
Technical field
The invention belongs to biomass cells and cultivate the reactor field, be applicable to that foodstuffs industry processing, agroforestry biological pesticide, biological products, bio-pharmaceuticals, biovaccine, high-density large-scale cells produce are cultivated and the cultivation of going down to posterity of biological medicine clone cell, be specially a kind of tidal type animal cell culture bioreactor.
Background technology
In the prior art, the cell cultures in agroforestry biological pesticide, biological products, bio-pharmaceuticals, biovaccine pharmaceutical factory is most cultivates in the incubator for placing with trial jar, is confined to the experiment type, type of production mostly is the Rotary Machine rolling bottle and cultivates.The Rotary Machine occupied ground is wide, input cost is big, production run expense height, cell yield are low, also be confined to the cultivation of cell category.
Summary of the invention
At the deficiencies in the prior art part: wide as the Rotary Machine occupied ground, input cost big, production run expense height, cell yield are low, be confined to deficiency that cell category cultivates etc.Type of the present invention aims to provide a kind of tidal type animal cell culture bioreactor, to overcome above-mentioned deficiency, makes that the cell cultivation equipment floor space is little, and cost is low, cell yield is high.
For achieving the above object, technical scheme of the present invention is:
A kind of tidal type animal cell culture bioreactor comprises culture medium supplemented or returnable bottle and heated constant temperature water tank, also comprises substratum fluid-containing pot and three gas water holding jacket constant incubators; Wherein, fixedly there are fluid-containing pot gas filter, pH value detecting electrode, sugar detecting electrode, dissolved oxygen detecting electrode, imbibition circulation tube a, feed liquor circulation tube a and additional liquid liquid-inlet pipe in the upper end of substratum fluid-containing pot; Be provided with the cell cultures retort that is filled with cell culture vector in the three gas water holding jacket constant incubators, one side of three gas water holding jacket constant incubators is provided with I peristaltic pump, II peristaltic pump and III peristaltic pump, and the cell cultures retort is provided with whipping appts, gas filter, level electrode, feed liquor circulation tube b and imbibition circulation tube b; Feed liquor circulation tube b is provided with the I reversing valve, and feed liquor circulation tube b is communicated with additional liquid pipette by return line, and return line is provided with the II reversing valve; The substratum fluid-containing pot is arranged in the heated constant temperature water tank or the outer water trapping layer of substratum fluid-containing pot is communicated with the heated constant temperature water tank by hot-water circulating pump and circulating pipe; The substratum fluid-containing pot is communicated with the cell cultures retort by imbibition circulation tube a, II peristaltic pump, feed liquor circulation tube b and I reversing valve; The cell cultures retort is communicated with the substratum fluid-containing pot by imbibition circulation tube a, III peristaltic pump and feed liquor circulation tube a; The substratum fluid-containing pot replenishes the liquid pipette by imbibition circulation tube a, II peristaltic pump, return line, II reversing valve, warp and is communicated with (this moment, the I reversing valve was to be in closing condition, and the II reversing valve is in open mode) with culture medium supplemented or returnable bottle; Culture medium supplemented or returnable bottle are communicated with (this moment, the II reversing valve was to be in closing condition) by replenishing liquid pipette, I peristaltic pump and additional liquid liquid-inlet pipe with the filling of substratum Sheng liquid.
The middle part of above-mentioned whipping appts is equipped with circulation tube air pressure interface, and the lower end of whipping appts is equipped with the internal recycle agitator.
Tidal type animal cell culture bioreactor of the present invention also comprises electrical equipment control cabinet and operation control display screen, and wherein the electrical equipment control cabinet has relay indicating light and power switch.
Tidal type animal cell culture bioreactor of the present invention also comprises falsework; Heated constant temperature water tank and culture medium supplemented or returnable bottle are placed in falsework lower floor, and three gas water holding jacket constant incubators are placed in the upper strata of falsework, and each member connects according to above-mentioned annexation.
Below the present invention will be further explained and the explanation:
One, the function of tidal type animal cell culture bioreactor each several part of the present invention is described in detail as follows:
1, three gas water holding jacket constant incubators (18): be to integrate water holding jacket constant temperature, carbonic acid gas, oxygen, the hybrid incubator of nitrogen, regulate carbonic acid gas, oxygen, nitrogen by three gas mixing variable valve (31), a kind of environment that is adapted to cell growth is provided, has guaranteed that effectively cell can high-density grow fast in the environment that its adapts to.
2, cell cultures retort (17): be filled with in the cell cultures retort (17) for cell attachment growing carrier (25), cell cultures retort (17) is provided with whipping appts (20), gas filter (22), level electrode (21) and feed liquor circulation tube b(12) and fluid circulation tube b(13).It is spongy porous fixed bed carrier that porous cell is cultivated carrier (25).
The cell cultures principle of cell cultures retort is: cell is sticked in the porous support of cultivating in the retort, substratum fluid-containing pot (3) is by imbibition circulation tube a(10), II peristaltic pump (15) and feed liquor circulation tube b(12) in cell cultures retort (17), carry substratum, make it be full of retort and flood carrier; And then by imbibition circulation tube b(13), III peristaltic pump (16) and fluid circulation tube a(11) be back in the substratum fluid-containing pot (3), to expose carrier; When carrier came out, cell can obtain sufficient oxygen and discharge carbonic acid gas.When carrier was flooded by substratum, cell can obtain nutrient and the waste of metabolism is discharged.Its principle shape is similar to the tide is flowing and ebbing.The design's device provides one with little space high-density cells, than the surface-area of maxicell growth, the environment of substratum non-foam, high oxygen supply in working cycle.Can be adapted to the cultivation of various kinds of cell.
3, fixedly there are fluid-containing pot gas filter (4), pH value detecting electrode (5), sugar detecting electrode (6), dissolved oxygen detecting electrode (7), imbibition circulation tube a(10 in the upper end of substratum fluid-containing pot (3)), feed liquor circulation tube a(11) and additional liquid liquid-inlet pipe (9); When needing results to reclaim liquid, close I reversing valve (33), open II reversing valve (44), reclaim liquid then in the substratum fluid-containing pot (3) by pipette a(10), II peristaltic pump (15), return line (45), II reversing valve (44) and culture medium supplemented pipette (8) flow in culture medium supplemented or the returnable bottle (30); When needing supplemental medium, close II reversing valve (44), the substratum in culture medium supplemented liquid or the returnable bottle (30) is by replenishing liquid pipette (8) through I peristaltic pump (14) and additional liquid liquid-inlet pipe (9) supplemental medium in the substratum fluid-containing pot (3); Realize reclaiming the function of liquid results or supplemental medium injection tank body.PH value detecting electrode (5), sugar detecting electrode (6) and dissolved oxygen detecting electrode (7) are for detection of the substratum each side index reactions change of fluid-containing pot.
4, heated constant temperature water tank (2): the water trapping layer that substratum fluid-containing pot (3) is arranged in the heated constant temperature water tank (2) or substratum fluid-containing pot (3) is outer is communicated with heated constant temperature water tank (2) by hot-water circulating pump (37) and circulating pipe (43), makes the interior temperature of the interior substratum preheating of substratum fluid-containing pot (3) and cell cultures retort (17) consistent.When in substratum circulation and the biological culture retort, difference variation is little, can make cell unaffected in culturing process.
5, the effect of culture medium supplemented or returnable bottle (30) is: when the substratum sugar degree in the substratum fluid-containing pot descends, can be by closing II reversing valve (44), the substratum in culture medium supplemented or the returnable bottle (30) is by replenishing liquid pipette (8) through I peristaltic pump (14) and additional liquid liquid-inlet pipe (9) supplemental medium in the substratum fluid-containing pot (3); Otherwise, when needing results to reclaim liquid, close I reversing valve (33), open II reversing valve (44), reclaim liquid then in the substratum fluid-containing pot (3) by pipette a(10), II peristaltic pump (15), return line (45) culture medium supplemented pipette (8) flow in culture medium supplemented or the returnable bottle (30).
6, I peristaltic pump (14), II peristaltic pump (15) and III peristaltic pump (16): act in this device, by control center control execution to the substratum in cell cultures retort and the substratum fluid-containing pot through the pipeline reciprocation cycle or replenish new substratum.
7, various pipelines: act on reciprocation cycle between the results of replenishing and reclaim liquid of new nutrient solution and substratum fluid-containing pot and the cell cultures retort.
8, display operation control panel (26): the whole device technique flow process of analog information and each location detection and red-tape operati intuitively.Tidal type animal cell culture bioreactor of the present invention is shown, operates and controlled by operation control display screen, shows the cultivation reaction of full device each several part technical process.The automatic allotment supply of ratio and the data of new culture medium supplemented are supported in detection cell cultures, substratum circulation, three air cultures, simulate dynamically to show respectively each cultivation reaction of installing of reactions change and control of device.The control intuitive display, simple to operation.
9, Controlling System cabinet (27): the data monitored, the collection and treatment that are each position in the whole device of control reach the conversion operation to each topworks.
10, relay indicating light (28): be each topworks's work indication in the whole device.
Two, the principle of tidal type animal cell culture bioreactor of the present invention:
Present design is that bio-reactor single unit system structure is placed in movably on the falsework (1), and compact construction is easy to operate.This reactor principle of work is: the cell cultures retort (17) of spongy porous being cultivated carrier (25) is inserted in the three gas water holding jacket constant incubators (18), the interior stationary temperature of three gas water holding jacket incubators (18) is set, mixes up three gas ratios and Water Tank with Temp.-controlled temperature simultaneously.Start is by II peristaltic pump (15), by imbibition circulation tube a(10) and feed liquor circulation tube b(12) substratum in the substratum fluid-containing pot (3) is conveyed in the cell cultures retort (17), the cell kind injected simultaneously.When substratum reached certain liquid level in the cell cultures retort (17), level electrode (21) control II peristaltic pump (15) stopped to carry substratum.Stirring motor this moment (20) starts internal recycle agitator (24), stirring more up turns clockwise at the bottom of making the interior substratum of cell cultures retort (17) by jar, the uniform overflow of the mouth of pipe comes and goes flows, and makes the well large-area all surfaces of porous support that are deposited to of cell kind.When internal recycle agitator (24) when quitting work, the rapid sedimentation of cell adheres on the carrier, and cell adhesion is behind carrier, and cell enters cultivation stage, cell cultures retort (17) beginning training.After cultivating for some time, cultivate substratum being sucked away at regular time and quantity in the retort, by imbibition circulation tube a(13), III peristaltic pump (16) and feed liquor circulation tube a(11) be back in the substratum fluid-containing pot (3), come and go circulation with cell cultures retort (17).When the substratum of cell cultures retort (17) was sucked away, the carrier cell that exposes obtained sufficient oxygen and discharges carbonic acid gas.When carrier was flooded by substratum, cell can obtain nutrient and the waste of metabolism is discharged.The electrode of substratum fluid-containing pot (3) detects the reactions change of every index of substratum constantly simultaneously, at any time in substratum fluid-containing pot (3), replenish new substratum, regulate the variation of substratum sugar, acid, basicity and regulate the interior gas concentration of three gas water holding jacket incubators (18).
Compared with prior art, the invention has the beneficial effects as follows:
Facility investment of the present invention is little, occupied ground is little, the production run cost is low, output is high.The output of one 5 liters tidal type biological culture reaction unit is equivalent to the rolling bottle of 25 bottles of positions, 3000ml/ bottle and cultivates 15 in machine, thereby the production platform of biological products and biological medicine and biovaccine effectively is provided, has reached purpose low-cost, that improve output.
Description of drawings
The invention will be further described below in conjunction with drawings and Examples
Fig. 1 is the one-piece construction synoptic diagram of the embodiment of the invention 2;
Fig. 2 is the heated constant temperature water tank (2) of Fig. 1 and the structural representation (band partial cutaway) of substratum fluid-containing pot (3);
Fig. 3 is the front view (band partial cutaway) of the three gas water holding jacket incubators (18) of Fig. 1;
Fig. 4 is the side-view (band partial cutaway) of the three gas water holding jacket incubators (18) of Fig. 1;
Fig. 5 is the structural representation (band partial cutaway) of the internal recycle agitator (24) of Fig. 1;
Fig. 6 is the one-piece construction synoptic diagram (band partial cutaway) of the embodiment of the invention 3;
Fig. 7 is the heated constant temperature water tank (2) of Fig. 6 and the structural representation (band partial cutaway) of substratum fluid-containing pot (3);
Wherein: 1 is falsework, 2 is the heated constant temperature water tank, 3 is the substratum fluid-containing pot, 4 is the fluid-containing pot gas filter, 5 is the pH value detecting electrode, 6 is the sugar detecting electrode, 7 is the dissolved oxygen detecting electrode, 8 for replenishing the liquid pipette, 9 for replenishing the liquid liquid-inlet pipe, 10 are imbibition circulation tube a, 11 is feed liquor circulation tube a, 12 is that feed liquor circulation tube b and 13 is imbibition circulation tube b, and 14 is the I peristaltic pump, and 15 is the II peristaltic pump, 16 is the III peristaltic pump, 17 are cell cultures reaction filling, and 18 is three gas water holding jacket constant incubators, and 19 is the outlet of internal recycle liquid, 20 is whipping appts, 21 is level electrode, and 22 is gas filter, and 23 is circulation tube air pressure interface, 24 is the internal recycle agitator, 25 are porous cell cultivation carrier, and 26 are operation control display screen, and 27 is the electrical equipment control cabinet, 28 is relay indicating light, 29 is power switch, and 30 for liquid medium replenishes or returnable bottle, and 31 is three gas hybrid regulators, 32 is carrier tray, 33 is the I reversing valve, and 34 is oxygen hose, and 35 is nitrogen tube, 36 is the internal recycle water screw, the 37th, hot-water circulating pump, the 38th, the comprehensive agitator of substratum, the 39th, glass vision panel in the fluid-containing pot, the 40th, stirring motor, the 41st, temperature detection sensor, the 42nd, constent temperature heater, 43 circulating pipes, 44 is the II reversing valve, and 45 is return line.
Embodiment
For a better understanding of the present invention, do detailed description further below in conjunction with the present invention of embodiment, but the scope of protection of present invention is not limited to the scope shown in the embodiment.Except specifying, the percentage composition in the Application Example of the present invention all refers to the quality percentage composition.
Embodiment 1:
A kind of tidal type animal cell culture bioreactor also comprises substratum fluid-containing pot (3) and three gas water holding jacket constant incubators (18); Wherein, fixedly there are fluid-containing pot gas filter (4), pH value detecting electrode (5), sugar detecting electrode (6), dissolved oxygen detecting electrode (7), imbibition circulation tube a(10 in the upper end of substratum fluid-containing pot (3)), feed liquor circulation tube a(11) and additional liquid liquid-inlet pipe (9); Be provided with the cell cultures retort (17) that is filled with cell culture vector (25) in the three gas water holding jacket constant incubators (18), one side of three gas water holding jacket constant incubators (18) is provided with I peristaltic pump (14), II peristaltic pump (15) and III peristaltic pump (16), and cell cultures retort (17) is provided with whipping appts (20), gas filter (22), level electrode (21), feed liquor circulation tube b(12) and imbibition circulation tube b(13); Feed liquor circulation tube b(12) be provided with I reversing valve (33), feed liquor circulation tube b(12) be communicated with additional liquid pipette (8) by return line (45), return line (45) is provided with II reversing valve (44); The water trapping layer that substratum fluid-containing pot (3) is arranged in the heated constant temperature water tank (2) or substratum fluid-containing pot (3) is outer is communicated with heated constant temperature water tank (2) by hot-water circulating pump (37) and circulating pipe (43); Substratum fluid-containing pot (3) is by imbibition circulation tube a(10), II peristaltic pump (15), feed liquor circulation tube b(12) and I reversing valve (33) be communicated with cell cultures retort (17); Cell cultures retort (17) is by imbibition circulation tube a(13), III peristaltic pump (16) and feed liquor circulation tube a(11) be communicated with substratum fluid-containing pot (3); Substratum fluid-containing pot (3) is by imbibition circulation tube a(10), II peristaltic pump (15), return line (45), II reversing valve (44), be communicated with (the I reversing valve is to be in closing condition at this moment, and the II reversing valve is in open mode) with culture medium supplemented or returnable bottle (30) through replenishing liquid pipette (8); Culture medium supplemented or returnable bottle (30) are communicated with (this moment, the II reversing valve was to be in closing condition) by replenishing liquid pipette (8), I peristaltic pump (14) and additional liquid liquid-inlet pipe (9) with substratum Sheng liquid filling (3).
As shown in Figure 5, the middle part of described whipping appts (20) is equipped with circulation tube air pressure interface (23), and the lower end of whipping appts (20) is equipped with internal recycle agitator (24).
A kind of tidal type animal cell culture bioreactor of the present invention also comprises electrical equipment control cabinet (27) and operation control display screen (26), and wherein electrical equipment control cabinet (27) has relay indicating light (28) and power switch (29).
Embodiment 2:
A kind of tidal type animal cell culture bioreactor of the present invention as shown in Figure 1, also comprises falsework (1); Heated constant temperature water tank (2) and culture medium supplemented or returnable bottle (30) are placed in falsework (1) lower floor, and three gas water holding jacket constant incubators (18) are placed in the upper strata of falsework (1), and each member connects according to embodiment 1 described annexation.
Embodiment 3:
A kind of tidal type animal cell culture bioreactor of the present invention, reactor as shown in Figure 6, wherein the outer water trapping layer of substratum fluid-containing pot (3) is communicated with heated constant temperature water tank (2) by hot-water circulating pump (37) and circulating pipe (43); (as shown in Figure 7), substratum fluid-containing pot (3) is placed in the next door of falsework, and other each member is identical with embodiment 1, connects according to embodiment 1 described annexation.
Embodiment 4: the working process of a kind of tidal type animal cell culture bioreactor of the present invention
The cell cultures retort (17) that will be placed with porous cell cultivation carrier (25) is inserted in the three gas water holding jacket constant incubators (18), and the interior stationary temperature of three gas water holding jacket incubators (18) is set, and mixes up three gas ratios and Water Tank with Temp.-controlled temperature simultaneously.Start is by II peristaltic pump (15), by imbibition circulation tube a(10) and feed liquor circulation tube b(12) substratum in the substratum fluid-containing pot (3) is conveyed in the cell cultures retort (17), the cell kind injected simultaneously.When substratum in the cell cultures retort (17) flooded whole porous cell cultivation carriers (25), level electrode (21) control II peristaltic pump (15) stopped to carry substratum.Start internal recycle agitator (24) this moment, stirrings that up turn clockwise again at the bottom of making substratum in the cell cultures retort (17) by jar, and the uniform overflow of the mouth of pipe comes and goes flows, and makes the cell kind can the good large-area all surfaces of porous support that are deposited to.When internal recycle agitator (24) when quitting work, enter cultivation stage behind the sticking wall of cell, cell cultures retort (17) begins to cultivate reaction work.After cultivating for some time, cultivate being sucked away of the interior substratum timing of retort, by imbibition circulation tube a(13), III peristaltic pump (16) and feed liquor circulation tube a(11) be back in the substratum fluid-containing pot (3), realize that substratum in the substratum fluid-containing pot (3) comes and goes with cell cultures retort (17) to circulate.When the substratum of cell cultures retort (17) was sucked away, the carrier cell that exposes obtained sufficient oxygen and discharges carbonic acid gas.When carrier was flooded by substratum, cell can obtain nutrient and the waste of metabolism is discharged.The electrode of substratum fluid-containing pot (3) detects the reactions change of every index of substratum constantly simultaneously, control center replenishes new substratum at any time in substratum fluid-containing pot (3), regulate the variation of substratum sugar, acid, basicity and regulate the interior gas concentration of three gas water holding jacket incubators (18).When needing results to reclaim liquid, close I reversing valve (33), open II reversing valve (44), reclaim liquid then in the substratum fluid-containing pot (3) by pipette a(10), II peristaltic pump (15), return line (45), II reversing valve (44) and culture medium supplemented pipette (8) flow in culture medium supplemented or the returnable bottle (30); When needing supplemental medium, close II reversing valve (44), the substratum in culture medium supplemented liquid or the returnable bottle (30) is by replenishing liquid pipette (8) through I peristaltic pump (14) and additional liquid liquid-inlet pipe (9) supplemental medium in the substratum fluid-containing pot (3); Realize reclaiming the function of liquid results or supplemental medium injection tank body.
Embodiment 6: Application Example:
1. with 5 liters of tidal type bioreactor culture PK15 cells:
1.1 vehicle treated: getting 100 gram cell culture vectors (25), is the PBS(deionized water preparation of 7.2,0.1M with pH value) 3 liters of immersions 2 hours.Naturally drain standby.
1.2 sterilization: take out cell cultures retort (17), add carrier and 2 liters of PBS of handling well, its all mouths of pipe bandage with clean-cloth, sterilize 40 minutes for 116 ℃ in Sterilizers.All mouths of pipe of substratum fluid-containing pot (3) bandage with clean-cloth, and articles for use such as silicone tube bandage with clean-cloth, sterilize 30 minutes for 121 ℃ in Sterilizers.
1.3 connecting tube: at hundred grades of clean environments, with aseptic technique cell cultures retort (17) outlet is connected with silicone tube with substratum fluid-containing pot (3) refluxing opening, cell cultures retort (17) fluid inlet is connected with silicone tube with substratum fluid-containing pot (3) outlet.
1.4 the parameter setting of cell cultures retort (17): the incubator temperature is 37 ℃, the volumetric concentration 5% of CO2.The rotating speed of whipping appts (20) is 150 rev/mins.The water temperature of substratum fluid-containing pot (3) is set at 37 ℃.
1.5 inoculating cell: open peristaltic pump the absorption of the PBS in the cell cultures retort (17) is discarded, add 2 liters and contain the PK15 kind cell that cell concn is 2 * 105/ml (10% new born bovine is clear with containing, and the high sugared DMEM substratum of two anti-100 units/ml are made into).Open internal recycle agitator (24) and stirred 30 minutes, after static 3 hours, restir 20 minutes stops to stir.
1.6 cell cultures: concrete culturing process is with embodiment 5, the time that the control carrier is flooded by substratum is 6min, siphoning away substratum then makes the time of cellular exposure also be 6min, recirculation like this, up to cell cultures retort (17) running about 96 hours, the glucose content of substratum is during less than 500mg/L, the substratum that need renew, and be covered with cell on the carrier this moment.
2. cultivate reactor with 5 liters of tidal type high-density carrier organism and cultivate PK15 cells produce swine Fever Vaccine:
2.1 cell cultures: with embodiment 5.
2.2 connect poison: behind the cell well-grown, discard old substratum then, it is clear that adding contains 4% new born bovine, two anti-100 units/ml, 5 liters of the high sugared DMEM substratum of swine fever spleen poison 0.5%.Cultivated about 72 hours, when glucose content was less than 500mg/L in the substratum, the results venom changed to that to contain 4% new born bovine clear, the high sugared DMEM substratum of two anti-100 units/ml.When glucose content is less than 500mg/L in substratum (about 72 hours), gathers in the crops venom, change substratum.
2.3 survey malicious result, see Table 1:
Table 1 PK15 cell cultures live vaccines of hog cholera test-results
Claims (6)
1. a tidal type animal cell culture bioreactor comprises culture medium supplemented or returnable bottle (30) and heated constant temperature water tank (2), it is characterized in that, also comprises substratum fluid-containing pot (3) and three gas water holding jacket constant incubators (18); Wherein, fixedly there are fluid-containing pot gas filter (4), pH value detecting electrode (5), sugar detecting electrode (6), dissolved oxygen detecting electrode (7), imbibition circulation tube a(10 in the upper end of substratum fluid-containing pot (3)), feed liquor circulation tube a(11) and additional liquid liquid-inlet pipe (9); Be provided with in the three gas water holding jacket constant incubators (18) and be filled with the cell cultures retort (17) that porous cell is cultivated carrier (25), one side of three gas water holding jacket constant incubators (18) is provided with I peristaltic pump (14), II peristaltic pump (15) and III peristaltic pump (16), and cell cultures retort (17) is provided with whipping appts (20), gas filter (22), level electrode (21), feed liquor circulation tube b(12) and imbibition circulation tube b(13); Feed liquor circulation tube b(12) be provided with I reversing valve (33), feed liquor circulation tube b(12) be communicated with additional liquid pipette (8) by return line (45), return line (45) is provided with II reversing valve (44); The water trapping layer that substratum fluid-containing pot (3) is arranged in the heated constant temperature water tank (2) or substratum fluid-containing pot (3) is outer is communicated with heated constant temperature water tank (2) by hot-water circulating pump (37) and circulating pipe (43); Substratum fluid-containing pot (3) is by imbibition circulation tube a(10), II peristaltic pump (15), feed liquor circulation tube b(12) and I reversing valve (33) be communicated with cell cultures retort (17); Cell cultures retort (17) is by imbibition circulation tube b(13), III peristaltic pump (16) and feed liquor circulation tube a(11) be communicated with substratum fluid-containing pot (3); Substratum fluid-containing pot (3) is by imbibition circulation tube a(10), II peristaltic pump (15), return line (45), II reversing valve (44), be communicated with culture medium supplemented or returnable bottle (30) through replenishing liquid pipette (8); Culture medium supplemented or returnable bottle (30) are communicated with substratum fluid-containing pot (3) by replenishing liquid pipette (8), I peristaltic pump (14) and additional liquid liquid-inlet pipe (9); It is spongy porous fixed bed carrier that described porous cell is cultivated carrier (25); The lower end of described whipping appts (20) is equipped with internal recycle agitator (24).
2. a kind of tidal type animal cell culture bioreactor according to claim 1 is characterized in that, the middle part of described whipping appts (20) is equipped with circulation tube air pressure interface (23).
3. a kind of tidal type animal cell culture bioreactor according to claim 1, it is characterized in that, also comprise electrical equipment control cabinet (27) and operation control display screen (26), wherein electrical equipment control cabinet (27) has relay indicating light (28) and power switch (29).
4. a kind of tidal type animal cell culture bioreactor according to claim 1 is characterized in that, also comprises falsework (1); Heated constant temperature water tank (2) and culture medium supplemented or returnable bottle (30) are placed in falsework (1) lower floor, and three gas water holding jacket constant incubators (18) are placed in the upper strata of falsework (1).
5. a kind of tidal type animal cell culture bioreactor according to claim 1 is characterized in that, the temperature of described substratum fluid-containing pot (3) is 37 ℃.
6. a kind of tidal type animal cell culture bioreactor according to claim 1, it is characterized in that, the temperature of described cell cultures retort (17) is 37 ℃, and the volumetric concentration of carbonic acid gas is 0-30%, and the rotating speed of whipping appts (20) is 0-250 rev/mins.
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| CN 201110387699 CN102382766B (en) | 2011-11-29 | 2011-11-29 | Tidal bioreactor for animal cell culture |
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| CN 201110387699 CN102382766B (en) | 2011-11-29 | 2011-11-29 | Tidal bioreactor for animal cell culture |
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| CN102382766A CN102382766A (en) | 2012-03-21 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102796661A (en) * | 2012-07-19 | 2012-11-28 | 南京福昌环保有限公司 | Anaerobic microbial biomass fermentation tank |
| CN103667053B (en) * | 2013-05-17 | 2015-07-29 | 中国航天员科研训练中心 | Automatic continuous changes liquid cell culture apparatus |
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| CN105985906B (en) * | 2015-02-04 | 2018-05-11 | 上海力申科学仪器有限公司 | High-temperature sterilization water-jacket typ incubator |
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| CN107513496A (en) * | 2016-06-17 | 2017-12-26 | 上海市农药研究所有限公司 | Unicellular alga auto culturing system and its application |
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| CN108641958B (en) * | 2018-08-08 | 2024-02-23 | 南京师范大学 | Biological three-dimensional printing environment real-time monitoring and activity maintaining device |
| CN109652296A (en) * | 2018-11-15 | 2019-04-19 | 上海量能生物科技有限公司 | The bioreactor of removable culture bottle pallet |
| CN109666583A (en) * | 2018-11-15 | 2019-04-23 | 上海量能生物科技有限公司 | Bioreactor with culture jar platform for placing |
| CN109628295A (en) * | 2018-11-15 | 2019-04-16 | 上海量能生物科技有限公司 | Bioreactor with culture jar pallet |
| CN111334427B (en) * | 2019-03-12 | 2023-05-05 | 华道(上海)生物医药有限公司 | Totally-enclosed cell culture system |
| CN111979088B (en) * | 2019-05-23 | 2022-08-26 | 比欧联科供应链管理(北京)有限公司 | Breathable bag forward and reverse perfusion cell culture device and culture method |
| CN110343607B (en) * | 2019-06-25 | 2020-09-22 | 南方医科大学珠江医院 | Biological reaction system for intermittent sampling |
| CN110423674B (en) * | 2019-08-19 | 2024-06-04 | 广东花猫农业科技有限公司 | Microorganism precipitation and propagation conveying device |
| CN111518667B (en) * | 2020-04-30 | 2021-05-04 | 苏州乐米凡电气科技有限公司 | Liquid medium replacing device with low potential pollution |
| CN113667590A (en) * | 2021-08-24 | 2021-11-19 | 品玺汇(厦门)生物科技有限公司 | Biological culture system |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101979517B (en) * | 2010-10-15 | 2011-12-28 | 普莱柯生物工程股份有限公司 | Method for producing influenza viruses in large scale by using bioreactor |
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