CN101260366A - Gas concentration and gas pressure adjustable cell culture case - Google Patents
Gas concentration and gas pressure adjustable cell culture case Download PDFInfo
- Publication number
- CN101260366A CN101260366A CNA2008100310708A CN200810031070A CN101260366A CN 101260366 A CN101260366 A CN 101260366A CN A2008100310708 A CNA2008100310708 A CN A2008100310708A CN 200810031070 A CN200810031070 A CN 200810031070A CN 101260366 A CN101260366 A CN 101260366A
- Authority
- CN
- China
- Prior art keywords
- box
- cell culture
- gas
- box body
- pressure
- 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.)
- Granted
Links
Images
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention provides a gas concentration and pressure adjustable type cell culture box, comprising a box body, a plastic bag communication pipe and a manometer communication pipe, wherein a side wall of the box body is provided with an air pipe connected with an air bag containing needed gases; the plastic bag communication pipe is communicated with a sealed plastic bag; the manometer communication pipe is communicated with a manometer; the lower part of a side wall of the box body is provided with a water pipe by which water can be drained out of the box or injected in the box, and the top part of the box body is provided with a box cover provided with a seal ring. The gas concentration and pressure adjustable type cell culture box has the advantages that: (1) mixed gases do not need to be pre-prepared, and different gases are directly injected into the cell culture box, which builds all gas conditions needed and saves gases and time, and the operation is easy; (2) the gas pressure in the cell culture box can be stably controlled, thereby the cell culture box can be used for the cell culture of which the gas pressure condition is the environmental atmospheric pressure as well as the research of the influence of the high gas pressure and the low gas pressure on cells; (3) the cell culture box matching a common constant temperature culture box can replace CO2 culture box, thereby saving costs.
Description
Technical field
The present invention relates to a kind of cell culture case, especially relate to the gentle pressure adjustable cell culture case of a kind of gas concentration.
Background technology
At present, when setting up cell culture condition, the common establishment method of cultivating the interior gas condition of vessel placement container (calling cell culture case in the following text) is, prepare earlier mixed gas on request, the mixed gas with preparation feeds the cultivation box again, replaces the atmosphere in the box, not only the gas waste is many, the poor controllability of gas concentration, and, the unbalanced problem of box internal gas pressure and around atmospheric pressure also ignored.Influence research for high atmospheric pressure or subatmospheric factor are cultivated cell in vitro also lacks accurately, the method for economy, simple and feasible at present.
Summary of the invention
The object of the present invention is to provide a kind of gas concentration controllability good, the cell culture case that air pressure is adjustable.
The objective of the invention is to be achieved through the following technical solutions: it comprises box body, on box side wall top ventpipe is installed, ventpipe connects the airbag that desired gas is housed, the box body top is provided with the lid of band sealing-ring, the box body outside have with box in the manometer that is communicated with, it is characterized in that, also be provided with the sealed plastic bag by being communicated with in communicating pipe and the box, the box side wall bottom is provided with the water pipe that is used for outside box draining or water filling in box.
Usually, laboratory the present invention's cell culture case, the vessel volume that is made of described box body is generally 3～5L, and the capacity of sealed plastic bag is generally 0.4～0.6L, and each ips can be 3～5mm.
Described sealed plastic bag volume can be adjusted automatically, thereby makes cultivation box internal gas pressure remain consistent with ambient air pressure.
All pipelines are preferably flexible pipe, also can select the hard tube of belt switch for use.
Described box body and pipe, airbag and related accessories are all made with non-toxic material.
The present invention's gas concentration controllability, the air pressure adjustability, mainly be based on following physics principle: (1) only communicates with the sack that desired gas is housed and keeps under the condition of air pressure balance and temperature-stable at the container that water is housed, discharge or inject in container the water of certain volume in the container, then the gas of equal volume enters or amount discharge container; (2) under the condition that gas-phase space remains unchanged in the container of water is housed, discharge or in container, inject the gas of certain volume in the container, can regulate the container internal gas pressure and reach certain level; (3) the utilization equation of gaseous state can calculate the interior total gas pressure of sealed vessel and the dividing potential drop and the volume fraction of all gases; (4), can calculate the gas dissolving amount in the certain volume water in the sealed vessel according to gas solubleness in water.
During use, operate according to the following steps: (1) adorns earlier an amount of water in described cell culture case, by the water of discharging certain volume equal-volume gas is entered in the box, makes the mixed gas of required all gases constituent concentration in box; (2), utilize the plastics bag statocyte that connects to cultivate the inside and outside air pressure of box if carry out the cell cultures that air pressure conditions is an around atmospheric pressure; (3),, regulate the cell culture case internal gas pressure and reach required level by injecting or extract out the gas of certain volume if study the influence of high atmospheric pressure and subatmospheric pair cell.
The invention has the advantages that: (1) does not need to prepare mixed gas in advance, directly feeds different gas in cell culture case, sets up required all gases condition, and saves gas and time, and is easy to operate; (2) can stably control the interior air pressure of cell culture case, both can be used for the cell cultures that air pressure conditions is an around atmospheric pressure, also can be used for studying the influence of high atmospheric pressure and subatmospheric pair cell; (3) this cell culture case cooperates common constant incubator energy replaced C O
2Incubator, but reduction of expenditure.
Description of drawings
Fig. 1 is the structural representation of one embodiment of the invention.
Embodiment
The invention will be further described below in conjunction with embodiment.
With reference to accompanying drawing, present embodiment comprises box body 12, in box body 12 side wall upper part ventpipe 8 is installed, ventpipe 8 connects the airbag 9 that desired gas is housed, box body 12 tops are provided with the lid 5 of band sealing-ring 7, box body 12 outsides have by the manometer 1 that is communicated with in communicating pipe 3 and the box, also are provided with the sealed plastic bag 2 by being communicated with in communicating pipe 4 and the box, and box body 12 lower sidewall are provided with the water pipe 10 that is used for outside box draining or water filling in box.All pipelines are flexible pipe, and the junction should be guaranteed air tight.
The vessel volume that is made of described box body 12 is 3L, and the capacity of sealed plastic bag 2 is 0.5L, and each inner diameter is 3mm.Box body and pipe, airbag and plastics bag, related accessories are all made with non-toxic material.
During use, should carry out according to following operation steps:
1. air pressure conditions is the cell cultures of around atmospheric pressure
(1) plastics bag communicating pipe 4, ventpipe 8 and water pipe 10 folders are closed, manometer communicating pipe 3 is communicated with manometer 1;
(2) open the lid 5, in the box that constitutes by box body 12, inject suitable quantity of water 13, culturing bottle 6 is placed on the carriage 11 that places in the box;
(3) cover lid 5,7 adjustment of the sealing-ring between lid 5 and the box mouth are compressed, guarantee that the box mouth is air tight;
(4) on ventpipe 8, connect the airbag 9 that desired gas is housed, unclamp the clip on the ventpipe 8, box body 12 is communicated with airbag 9;
(5) unclamp clip on the water pipe 10, allow the water in the box flow in the graduated cylinder, the gases in the airbag 9 enter in the box simultaneously; Note observing manometer 1, the box internal gas pressure should keep balancing each other with around atmospheric pressure; When the volume (promptly entering the gas volume box in) that flows out water when reaching desired value, folder closes water pipe 10 immediately, and folder closes ventpipe 8 and takes off dress airbag 9 subsequently, presses following formula (1) calculating charge volume:
V＝aDV
2-bV
1+(aC
3-bC
1)V
w3T
1/273 (1)
In the formula,
V: certain gas feeds required volume in the box;
A: the percentage concentration of this gas (volume fraction) during cultivation;
D: the compensation water vapour pressure causes gas-phase space in the box to increase and keeps the balance each other coefficient of required this gas feeding amount of box internal gas pressure and around atmospheric pressure, D=(P
1-P
W2)/(P
1-P
W3),
Wherein, P
1Box internal gas pressure (=around atmospheric pressure) during for ventilation, P
W2: the saturated vapor pressure under the temperature in box during ventilation, P
W3: the saturated vapor pressure under the temperature in box during cell cultures;
V
2: finish the volume of the interior gas-phase space of box when feeding all gases, V
2=V
0-V
W2-V
c,
Wherein, V
0For cultivating box cubic capacity, V
W2For retaining the volume of water, V in the box
cFor cultivating vessel, culture system and carriage volume sum;
B: the percentage concentration (volume fraction) of this gas in atmosphere;
V
1: atmosphere gas-phase space volume in the box of setting before the ventilation, V
1=V
0-V
W1-V
c,
Wherein, V
W1For presetting the volume of water in the box before the ventilation;
C
1: the solubleness of this gas in water under the interior temperature of box during ventilation;
C
3: the solubleness of this gas in water under the interior temperature of box during cultivation;
V
W3: retain the volume of water during cultivation in the box, V
W3=V
W2
T
1: the absolute temperature (K) during ventilation in the box;
(6) in cell culture case, feed the 2nd kind of gas, then repeating step (5) as need; So analogize, feed more kinds of gases;
(7) after feeding had all gases of concentration requirement, still water fed nitrogen in the box in box by emitting, and made the interior volume of water of retaining of box reduce to set(ting)value V
W3Nitrogen air input V
NPressing following formula (2) calculates:
V
N＝V
0-V
1-V
c-V
w3-V
s (2)
In the formula,
V
s: except that the nitrogen that enters at last, all gases volume sum of other feeding.
The nitrogen speed that feeds should be slower, when nitrogen feeding amount near V
NThe time (retain volume of water in the box near V
W3The time), folder closes ventpipe 8 and water service pipe 10, stops to ventilate several minutes, so that the water vapor pressure that reaches capacity in the box; Observe the variation of box internal gas pressure by manometer 1,, unclamp the clip on the water pipe 10 earlier, slowly emit less water, when in box, pressing, unclamp the clip on the ventpipe 8, make nitrogen feeding amount reach V with the around atmospheric pressure balance when pressing in the box when stopping to rise
N, folder closes ventpipe 8 and water pipe 10 immediately;
(8) folder stopped pipe 3 takes off manometer 1;
(9) unclamp clip on the pipe 4, make that a gasless sealed plastic bag 2 communicates in box 12 and the bag;
(10) culturing bottle 6 is put into common constant incubator, carry out cell culture experiments; Temperature raises, and gas expansion in the box, portion gas just can enter gasless sealed plastic bag 2 originally, makes that normal atmosphere remains consistent in box internal gas pressure and the culture environment.
2. air pressure conditions is higher or lower than the cell cultures of around atmospheric pressure
Step (1)～(6) are identical with " 1. air pressure conditions is the cell cultures of around atmospheric pressure ".But press the air input that following formula (3) calculates the projecting normal atmosphere cell cultures of air pressure conditions, calculate the air input that air pressure conditions is lower than the around atmospheric pressure cell cultures by following formula (4):
V＝aV
2-bV
1+(aC
3-bC
1)V
w3T
1/273 (3)
V＝aQV
2-bV
1+(aC
3-bC
1)V
w3T
1/273 (4)
In the formula (4),
Q: compensation causes the coefficient that desired gas reduces in the box, Q=(P because of bleeding
2-P
W2)/(P
i-P
W2),
Wherein, P
2The pressure of gas-phase space in the box when feeding all gases that concentration requirement is arranged for finishing, P
jThe level [pressing following formula (5) calculates] that the reduction of box internal gas pressure will be reached;
(7) with syringe by ventpipe 8 nitrogen injection or withdrawing gas in the box in box, the box internal gas pressure raise or reduce, the level that this moment, the box internal gas pressure will reach (by manometer 1 monitoring) press following formula (5) calculating:
P
i＝(P
3-P
w3)T
2/T
3+P
w2 (5)
In the formula,
P
i: nitrogen injection or withdrawing gas raise the box internal gas pressure or reduce the level that will reach.
P
3: the air pressure level that requires in the box during cell cultures.
T
2: cultivate the kelvin rating (=T in the box during ventilation
1).
T
3: the kelvin rating during cultivation in the box.
(8) folder closes ventpipe 8, takes off syringe; Folder stopped pipe 3 takes off manometer 1;
(9) culturing bottle 6 is put into common constant incubator, carry out cell culture experiments.
Claims (2)
1. gentle pressure adjustable cell culture case of gas concentration, comprise box body, on box side wall top ventpipe is installed, ventpipe connects the airbag that desired gas is housed, the box body top is provided with the lid of band sealing-ring, the box body outside have with box in the manometer that is communicated with, it is characterized in that, also be provided with the sealed plastic bag by being communicated with in communicating pipe and the box, the box side wall bottom is provided with the water pipe that is used for outside box draining or water filling in box.
2. the gentle pressure adjustable cell culture case of gas concentration according to claim 1 is characterized in that, the vessel volume that is made of described box body is 3～5L, and the capacity of sealed plastic bag is 0.4～0.6L, and each ips is 3～5mm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008100310708A CN101260366B (en) | 2008-04-15 | 2008-04-15 | Gas concentration and gas pressure adjustable cell culture case |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008100310708A CN101260366B (en) | 2008-04-15 | 2008-04-15 | Gas concentration and gas pressure adjustable cell culture case |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101260366A true CN101260366A (en) | 2008-09-10 |
| CN101260366B CN101260366B (en) | 2011-04-06 |
Family
ID=39961066
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2008100310708A Expired - Fee Related CN101260366B (en) | 2008-04-15 | 2008-04-15 | Gas concentration and gas pressure adjustable cell culture case |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101260366B (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102660459A (en) * | 2012-05-14 | 2012-09-12 | 韩宁 | Anaerobic culture box and using method thereof |
| CN104162372A (en) * | 2013-08-21 | 2014-11-26 | 湖南师范大学 | Gas quantitative blending and speed-controlled transmission method and device based on water-gas exchange technology |
| CN104565808A (en) * | 2014-12-26 | 2015-04-29 | 湖南师范大学 | Gas replacement method and device used for rigid container and based on quantitative gas extracting-inflating technology |
| CN104673628A (en) * | 2015-02-16 | 2015-06-03 | 陈子江 | Heat-preservation culturing device with gas control function |
| CN105866275A (en) * | 2016-04-05 | 2016-08-17 | 济宁神农画圆生态***模式推广有限公司 | Method and device for quantitative analysis on nitrogen-oxygen absorption capacity of nitrogen-fixing bacteria in sealed container |
| CN107012088A (en) * | 2017-05-12 | 2017-08-04 | 中国人民解放军总医院 | A kind of adjustable bacteriological incubator of air pressure |
| CN108467834A (en) * | 2018-02-11 | 2018-08-31 | 温州医科大学 | A kind of small-sized cell culture unit of pressure controllable |
| CN109929759A (en) * | 2019-03-22 | 2019-06-25 | 山东农业大学 | A kind of acquisition of egg mother cell and early stage molding culture apparatus |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2151154Y (en) * | 1992-08-13 | 1993-12-29 | 王守亮 | Cycling cell cultivator |
| CN2608509Y (en) * | 2003-03-13 | 2004-03-31 | 李伟 | Microbiological reactor |
| CN1858203A (en) * | 2006-04-05 | 2006-11-08 | 中国人民解放军第三军医大学第一附属医院 | Pneumatically controlled pressurizing cell cultivation instrument |
-
2008
- 2008-04-15 CN CN2008100310708A patent/CN101260366B/en not_active Expired - Fee Related
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102660459A (en) * | 2012-05-14 | 2012-09-12 | 韩宁 | Anaerobic culture box and using method thereof |
| CN104162372A (en) * | 2013-08-21 | 2014-11-26 | 湖南师范大学 | Gas quantitative blending and speed-controlled transmission method and device based on water-gas exchange technology |
| CN104162372B (en) * | 2013-08-21 | 2016-05-18 | 湖南师范大学 | Gasometry mixture based on water-gas replacement technique and the fast carrying method of control and device |
| CN104565808A (en) * | 2014-12-26 | 2015-04-29 | 湖南师范大学 | Gas replacement method and device used for rigid container and based on quantitative gas extracting-inflating technology |
| CN104673628A (en) * | 2015-02-16 | 2015-06-03 | 陈子江 | Heat-preservation culturing device with gas control function |
| CN105866275A (en) * | 2016-04-05 | 2016-08-17 | 济宁神农画圆生态***模式推广有限公司 | Method and device for quantitative analysis on nitrogen-oxygen absorption capacity of nitrogen-fixing bacteria in sealed container |
| CN107012088A (en) * | 2017-05-12 | 2017-08-04 | 中国人民解放军总医院 | A kind of adjustable bacteriological incubator of air pressure |
| CN107012088B (en) * | 2017-05-12 | 2023-11-24 | 中国人民解放军总医院 | Air pressure adjustable bacterial incubator |
| CN108467834A (en) * | 2018-02-11 | 2018-08-31 | 温州医科大学 | A kind of small-sized cell culture unit of pressure controllable |
| CN108467834B (en) * | 2018-02-11 | 2023-07-25 | 温州医科大学 | Pressure-controllable small cell culture device |
| CN109929759A (en) * | 2019-03-22 | 2019-06-25 | 山东农业大学 | A kind of acquisition of egg mother cell and early stage molding culture apparatus |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101260366B (en) | 2011-04-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101260366B (en) | Gas concentration and gas pressure adjustable cell culture case | |
| CN201737949U (en) | Cell culturing device capable of periodically converting positive pressure and negative pressure | |
| CN205099688U (en) | Hypoxemia environment carbon dioxide cell culture case | |
| CN203530286U (en) | White spirit solid state fermentation device for laboratory | |
| CN104450519B (en) | The cell culture system of a kind of precise hard_drawn tuhes nutrient solution dissolved oxygen concentration | |
| CN204474690U (en) | The cell culture system of precise hard_drawn tuhes nutrient solution dissolved oxygen concentration | |
| CN101480135B (en) | Device for cultivating liquid culture of edible fungus | |
| CN103173347B (en) | Multifunctional continuous fermentation and extraction integrated apparatus | |
| CN204198730U (en) | Anerobe, microaerobe and need carbonic acid gas bacterium culture bag | |
| CN207376060U (en) | A kind of anaerobe domesticating device | |
| CN203653553U (en) | Real-time sampling in-vitro culture fermentation bottle | |
| CN206706123U (en) | A kind of cell culture bags | |
| CN202925194U (en) | Semi-automatic all-in-one machine for down feeding and down filling | |
| CN211311449U (en) | Silage maize fermenting installation | |
| CN201176433Y (en) | Apparatus preparing anaerobic culture fluid | |
| CN204918603U (en) | Cell co -culture system under pressure environment | |
| CN209537509U (en) | A kind of high leakproofness 500mL plastic culture base square vase | |
| CN203960185U (en) | A kind of anaerobic fermentation system | |
| CN208733088U (en) | Sample rate separating liquid save set | |
| GEP20196992B (en) | Improved fermenter | |
| CN206851649U (en) | A kind of accurate quantitative filling device of hand | |
| CN206599566U (en) | Use for laboratory is dry, the integrated apparatus of wet-fermentation methane phase | |
| CN206970621U (en) | A kind of inner membrance air storing type integrated methane fermentation device | |
| CN204550627U (en) | A kind of air ventilation device for cell cultures | |
| CN104031833A (en) | Clutch gravity type fermentation and extraction integrated tank and extraction method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20110406 Termination date: 20120415 |