CN102329724A - System and method for testing influence of pressure on cell growth - Google Patents
System and method for testing influence of pressure on cell growth Download PDFInfo
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- CN102329724A CN102329724A CN201110283487A CN201110283487A CN102329724A CN 102329724 A CN102329724 A CN 102329724A CN 201110283487 A CN201110283487 A CN 201110283487A CN 201110283487 A CN201110283487 A CN 201110283487A CN 102329724 A CN102329724 A CN 102329724A
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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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/40—Means for regulation, monitoring, measurement or control, e.g. flow regulation of pressure
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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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/08—Flask, bottle or test tube
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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
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/20—Degassing; Venting; Bubble traps
Abstract
The invention relates to a system and method for testing influence of pressure on cell growth. The system comprises a pressure box, a culture flask filled with cells, a gas steel cylinder filled with gas, a pressure meter, a first scavenging valve, a second scavenging valve, a first gas conveying pipe and a second gas conveying pipe, wherein the pressure box comprises a shell, the culture flask is arranged inside the shell, the shell is provided with a gas inlet, an exhaust hole and a pressure detection hole which enable the interior and exterior of the shell to be communicated, the outlet of the first scavenging valve is connected with the gas inlet, the inlet of the first scavenging valve is connected with the gas steel cylinder through the first gas conveying pipe, the inlet of the second scavenging valve is connected with the exhaust hole, one end of the second gas conveying pipe is connected with the pressure meter, and the other end of the second scavenging valve is communicated with the inside of the shell through the pressure detection hole. According to the invention, the influence of a preset test pressure on the cell growth for a long time can be accurately tested.
Description
Technical field
The present invention relates to the human body cell technical field of measurement and test, particularly relate to a kind of system and method for influence of test pressure cell growth.
Background technology
Human body has a variety of cells, as retinal ganglial cells, tm cells etc. are arranged in the eyes, endotheliocyte etc. is arranged in cardiovascular.The various cells of human body are all lived under certain pressure environment, and the variation of this pressure impacts the growth of pair cell.For example, glaucoma (Glaucoma) is global deputy non-reversibility diseases causing blindness, and data shows according to statistics, and the whole world has 7,000 ten thousand glaucoma patients approximately, 6,700,000 blindings wherein, and China has 7,000,000 glaucoma patients approximately, sickness rate is about 0.52%.These sick characteristics are that intraocular pressure raises, and optic nerve is impaired, also with carrying out property defect of visual field.At present efficacious therapy means are that the method through medicine, laser or operation controls to normal level with intraocular pressure, thereby alleviate or suppress the damage of pressure to optic nerve.Again for example, hypertension can cause damage to cardiovascular endotheliocyte, thereby causes various cardiovascular disordeies, perhaps brings out heart trouble.Also be to utilize medicine to control patient's blood pressure to hypertensive treat-ment at present, make it be no more than the normal range of 120mmHg (mmhg)/80mmHg.
But, no matter be of the infringement of high intraocular pressure at present, or hypertension is to the damage of cardiovascular endotheliocyte to intraocular cell (like retinal ganglial cells, tm cells etc.), its pathomechanism is also not fully aware of.In order to study this pathomechanism, just need corresponding apparatus to come the influence of test pressure cell growth.The system construction drawing of the influence of the test pressure cell growth that Fig. 1 provides for prior art.As shown in Figure 1; This test is carried out in a culturing bottle 101, place cell to be studied 104 in the culturing bottle 101 after, seal the bottleneck 102 of this culturing bottle 101 with certain material (like plug etc.); Make it inside and outside isolated; Only there is syringe 103 so that culturing bottle 101 pressure inside reach predetermined test pressure, to begin test then to the delivered inside air of this culturing bottle 101.When the test duration reaches the scheduled time, open bottleneck 102, cell 104 is taken out to analyze and research from culturing bottle 101 get final product.
The cell of testing with this system 104 is active somatic cells, thereby it must carry out the exchange of material with the external world, absorbs the oxygen in the culturing bottle 101 thereby respiration for example will take place, and produces carbonic acid gas and be expelled in the culturing bottle 101.And generally; The volume of culturing bottle 101 very little (about 50ml), after test after a while, bigger variation can take place in the gaseous constituent in this culturing bottle 101; At this moment the atmosphere surrounding in this culturing bottle 101 just wants mimic human body environment far different with it; Thereby the result of this test also just comprised the influence of atmosphere surrounding factor pair cell, can not correctly reflect the influence of pure pressure (like high pressure) pair cell, and is visible; Prior art can only the simple test pressure pair cell short period of time (in several minutes) impact effect, and meaning that should test and little.In addition; Prior art utilizes syringe 103 in culturing bottle 101, to inject air, thereby makes the air pressure in the culturing bottle 101 reach predetermined test pressure, and this technology not only wastes time and energy; And can not accurately judge the pressure condition in the culturing bottle 101, thereby the result of test also is more coarse.
Summary of the invention
Technical problem to be solved by this invention provides a kind of system and method for influence of test pressure cell growth, can accurately test the influence of long-time interior presumptive test pressure cell growth.
The technical scheme that the present invention solves the problems of the technologies described above is following: a kind of system of influence of test pressure cell growth, and this system comprises: culturing bottle, the gas bomb that gas is housed, tensimeter, a scavenging air valve, No. two scavenging air valves, an air shooter and No. two air shooters of pressure tank, the said cell of loading; Wherein, said pressure tank comprises shell;
Said culturing bottle is positioned at said enclosure;
Have on the said shell and make air inlet port, venting hole and the pressure detection hole that is interlinked to the outside in it;
The outlet of a said scavenging air valve connects said air inlet port, and it enters the mouth through a said said gas bomb of gas delivery pipe coupling;
The inlet of said No. two scavenging air valves connects said venting hole;
One end of said No. two air shooters links to each other with said tensimeter, and the other end communicates with said enclosure through said pressure detection hole.
The invention has the beneficial effects as follows: among the present invention; Culturing bottle is positioned at the shell of pressure tank; Gas bomb links to each other with the inlet of the scavenging air valve that air inlet port was connected of shell through an air shooter; Can when a scavenging air valve is in opened condition, gas be provided,, and make the air pressure of enclosure reach predetermined test pressure with the gas of displacement enclosure to enclosure; The gas of enclosure can be discharged through No. two scavenging air valves that venting hole connected, and the air pressure of enclosure (being the pressure that cell bore in the culturing bottle) can accurately be read by tensimeter.Like this; The space of the enclosure among the present invention is more much bigger than the volume of the culturing bottle about 50ml in the prior art; Even cell is through long respiration; The gas composition of enclosure is taken place than about-face, thereby test result of the present invention can accurately reflect the influence of long-time internal pressure factor for the cell growth.The present invention utilizes tensimeter, can accurately obtain the air pressure in the pressure tank shell, coarse with respect to prior art, and the present invention is accurately for the setting of test pressure, thereby test result also is accurately convictive.
On the basis of technique scheme, the present invention can also do following improvement:
Further, the thermostat container that also comprises the steady temperature that inner maintenance is preset;
Said pressure tank is positioned at the inside of said thermostat container.
Further, said pressure tank also comprises internal partition, and its level is positioned at the predetermined height of the top, bottom surface of said shell, and said enclosure is divided into superposed culturing room and the damp camera that is positioned at the bottom;
Have more than one through hole on the said internal partition, said damp camera and said culturing room are communicated;
Said culturing bottle is positioned at said culturing room;
This system also comprises the absorbent cotton that is soaked with sterile distilled water, and it is positioned at said damp camera.
Further, said pressure tank also comprises circular turn switch door, and it is fitted through plastic pad and said case surface.
Further, also comprise the gas concentration unit that links to each other with the outlet of said No. two scavenging air valves.
In addition, the present invention also provides a kind of method of influence of test pressure cell growth, and this method is based on above-mentioned system; This method comprises:
Step 1: the culturing bottle that will load said cell places the enclosure of said pressure tank;
Step 2: open a said scavenging air valve and said No. two scavenging air valves; Open said gas bomb, make gas wherein arrive said enclosure, the original gas of said enclosure is discharged from said No. two scavenging air valves along a said air shooter;
Step 3: after the original gas of said enclosure all is discharged to said housing exterior, close said No. two scavenging air valves;
Step 4: when said manometric registration reaches predetermined pressure, close a said gas bomb and a said scavenging air valve successively;
Step 5: when the test duration arrives the scheduled time, open said No. two scavenging air valves, the gas of said enclosure is discharged from said No. two scavenging air valves;
Step 6: take out said culturing bottle from said enclosure, the said cell of its loading is analyzed.
Further, after said step 1, before said step 2, also comprise step 1-1: said pressure tank is placed in the thermostat container, and make said thermostat container temperature inside remain preset steady temperature.
Further, the enclosure of said pressure tank is divided into superposed culturing room and is positioned at the damp camera of bottom by internal partition, and has more than one through hole on the said internal partition so that said damp camera and said culturing room communicate; Then
Said step 1 is: the culturing bottle that will load said cell places in the culturing room of enclosure of said pressure tank;
Said step 1 also comprises: the absorbent cotton that will be soaked with sterile distilled water places in the said damp camera, so that the humidity of said enclosure reaches predetermined moisture.
Further, before said step 1, also comprise step 0: the turn switch door of opening said pressure tank;
After said step 1, before said step 2, also comprise step 1-2: close said turn switch door, and its plastic pad and said case surface are fitted.
Further, the gas in the said gas bomb is: nitrogen; Or, carbon monoxide; Or, contain volume ratio and be the air of 5% carbonic acid gas.
Description of drawings
The system construction drawing of the influence of the test pressure cell growth that Fig. 1 provides for prior art;
Fig. 2 is the system construction drawing of the influence of test pressure cell growth provided by the invention;
Fig. 3 is the structure iron of culturing bottle provided by the invention;
Fig. 4 is the schema of method of the influence of test pressure cell growth provided by the invention.
Embodiment
Below in conjunction with accompanying drawing principle of the present invention and characteristic are described, institute gives an actual example and only is used to explain the present invention, is not to be used to limit scope of the present invention.
Fig. 2 is the system construction drawing of the influence of test pressure cell growth provided by the invention; As shown in Figure 2, this system comprises: pressure tank 201, the culturing bottle that loads cell, the gas bomb 202 that gas is housed, tensimeter 203, scavenging air valve 205, No. two scavenging air valves 206, an air shooter 204 and No. two air shooters 207; Pressure tank 201 wherein comprises shell 2011; A scavenging air valve 205 and No. two scavenging air valves 206 can use common scavenging air valve (like the scavenging air valve of natural gas line) to realize; An air shooter 204 and No. two air shooters 207 can be realized with the pipeline that vulcanite is made, thereby guarantee resistance to air loss.
Culturing bottle is the device that carries cell, and it is positioned at shell 2011 inside.Among the present invention; The structure of culturing bottle is as shown in Figure 3, comprises bottle 301 and filter membrane 302, and this bottle 301 is for loading the container that only has an opening of cell 303; Its structure is varied; For example be the cylindrical semienclosed container of open-topped hollow shown in Figure 3, certainly, also can be rectangular parallelepiped semienclosed container, square container or the erose container of open-topped hollow.Cell 303 is packed into behind bottle 301 inside; To bottleneck be sealed with filter membrane 302; Get in the bottle 301 with the prevention bacterium, but keep gas to free in and out bottle 301, thereby the filter membrane among the present invention 302 has micro pores through filter membrane 302; The diameter of its hole is less than the diameter of bacterium, and greater than the diameter of gas of being adorned in the gas bomb 202 (like nitrogen, carbon monoxide, air etc.) and water vapor.In addition, packing into before the cell 303 in bottle 301 inherences, can also sterilize fully.
Have on the shell 2011 to make air inlet port 20111, venting hole 20112 and the pressure detection hole 20113 that is interlinked to the outside in it, thereby these three holes are the form of through hole.
The outlet of a scavenging air valve 205 connects air inlet port 20111; The inlet of a scavenging air valve 205 connects gas bomb 202 through an air shooter 204; Like this, when a scavenging air valve 205 is in opened condition, can open gas bomb 202; Make its gas inside get into a scavenging air valve 205 along an air shooter 204; And get into shell 2011 inside from the outlet of a scavenging air valve 205, and the entering of these gases has two kinds of effects: the one, and shell 2011 inner original gases (like air and with the used gas of Pretesting) are replaced, thereby guarantee the pure of shell 2011 gas inside in this test; The 2nd, improve the air pressure of shell 2011 inside, make it reach predetermined test pressure.
Shell 2011 inner air pressure are to utilize the tensimeter 203 among Fig. 2 to detect, and as shown in Figure 2, an end of No. two air shooters 207 links to each other with tensimeter 203, and the other end communicates with shell 2011 inside through pressure detection hole 20113.Like this, the present invention utilizes tensimeter 203 can obtain the accurate air pressure of shell 2011 inside, makes test pressure be predetermined force value, guarantees the accuracy and the cogency of test.And in the prior art, because culturing bottle 101 bottle packaging air tightness, the detected result of air pressure is inaccurate.
The inlet of No. two scavenging air valves 206 connects venting hole 20112; Its outlet can communicate with outside air; Certainly; Because the gases in the gas bomb 202 possibly be toxic gases such as carbon monoxide, thereby the outlet of No. two scavenging air valves 206 here can connect gas concentration unit (like balloon, gas recovery bag etc.), thereby guarantees tester's safety.In addition, can also carry out composition analysis, thereby further judge the variation that shell 2011 gas inside environment are taken place, for test accuracy of the present invention provides foundation the gas in the gas concentration unit.
This shows; Among the present invention; Culturing bottle is positioned at the shell of pressure tank, and gas bomb links to each other with the inlet of the scavenging air valve that air inlet port was connected of shell through an air shooter, can when a scavenging air valve is in opened condition, to enclosure gas be provided; Gas with the displacement enclosure; And the air pressure that makes enclosure reaches predetermined test pressure, and the gas of enclosure can be discharged through No. two scavenging air valves that venting hole connected, and the air pressure of enclosure (being the pressure that cell bore in the culturing bottle) can accurately be read by tensimeter.Like this; The space of the enclosure among the present invention is more much bigger than the volume of the culturing bottle about 50ml in the prior art; Even cell is through long respiration; The gas composition of enclosure is taken place than about-face, thereby test result of the present invention can accurately reflect the influence of long-time internal pressure factor for the cell growth.The present invention utilizes tensimeter, can accurately obtain the air pressure in the pressure tank shell, coarse with respect to prior art, and the present invention is accurately for the setting of test pressure, thereby test result also is accurately convictive.
Certainly, above-mentioned gas concentration unit also can be replaced with gas-filtering device, and this gas-filtering device is used for filtering some composition (like toxic gases such as carbon monoxide) from No. two scavenging air valve 206 expellant gas, thereby guarantees tester's safety.
In order to guarantee the resistance to air loss of shell 2011; These three positions, the outside of the outside of the inlet link position of the outside of the outlet link position of an air inlet port 20111 and a scavenging air valve 205, venting hole 20112 and No. two scavenging air valves 206 and pressure detection hole 20113 and No. two air shooter 207 link positions can also be provided with glass cement on shell 2011; To guarantee the airtight good of air inlet port 20111, venting hole 20112,20113 places, pressure detection hole, prevent that outside air from getting into shell 2011 inside.
In order to improve the degree of closeness of test and human body environment, this test can also be carried out under temperature constant state, thereby this system can also comprise thermostat container; The inside of this thermostat container can remain preset steady temperature; Like this, just can pressure tank be arranged at the inside of thermostat container, regulate the thermostat container temperature inside; Make its temperature that remains position in the cell 303 place human bodies, thereby improve the accuracy of this test.
Realize putting into and taking out of culturing bottle; An opening just need be set on the shell 2011 of pressure tank 201, a chamber door is set on this opening, as shown in Figure 2; Pressure tank 201 comprises that also circular turn switch door 2012 is as chamber door; This turn switch door 2012 is fitted through the surface of plastic pad and shell 2011, thereby guarantees the resistance to air loss of this opening part, prevents that extraneous air from passing through opening entering shell 2011 inside that this turn switch door 2012 is sealed.Turn switch door 2012 is that the mode of the switch that is provided with on it through turn realizes the compressing of plastic pad, thereby the resistance to air loss of this chamber door is very good.
As shown in Figure 2; Pressure tank 201 also comprises internal partition 2013 except comprising shell 2011, turn switch door 2012, its level is positioned at the predetermined height (being certain position between the upper and lower surfaces of shell 2011) of the bottom surface top of shell 2011; Shell 2011 inside are divided into superposed culturing room and the damp camera that is positioned at the bottom; Shell 2011 shown in Figure 2 is a square, and internal partition 2013 wherein then is the length of side and shell 2011 interior edge appearance square together, and its edge fits tightly on the inner surface of shell 2011; Have the through hole 20131 of (Fig. 2 shows has 3) more than on this internal partition 2013, damp camera and culturing room are communicated.
Can culturing bottle be placed on the upper surface of internal partition 2013, be about to it and place in the culturing room, and keep the unobstructed of through hole 20131, the absorbent cotton that is soaked with sterile distilled water is set in addition, be placed on the bottom surface in the shell 2011, be about to it and place in the damp camera.Like this, guarantee shell 2011 humidity inside, make its humidity that reaches cell 303 place position of human body, thereby improve this accuracy of test with regard to the sterile distilled water that absorbent cotton capable of using soaked.Here, the big I of humidity is regulated the detection of humidity wet bulb thermometer capable of using through the mode that changes absorbent cotton quantity in the shell 2011.
Certainly, culturing bottle is being placed shell 2011 inside (in the culturing room) before, also can carry out strict sterilization shell 2011 inside.
The internal partition 2013 here might not be illustrated in figure 2 as one, can also be for more than two, and each internal partition is horizontally placed on the different heights place of shell 2011 inside; Thereby shell 2011 inside are divided into a plurality of independent subspaces; Be equipped with more than one through hole on each internal partition, communicate with other subspaces keeping, (as the subspace below being positioned at, one of them (or a plurality of) subspace; Or the subspace in the middle part of being positioned at; Or superposed subspace) as damp camera, be used to be provided with the absorbent cotton that is soaked with sterile distilled water, other subspace is as culturing room; Be used to be provided with culturing bottle, thereby can carry out the test of various kinds of cell under same test pressure, same pressure.
Based on above-mentioned system, the invention allows for a kind of method of influence of test pressure cell growth, Fig. 4 is the schema of this method.As shown in Figure 4, this method comprises:
Step 401: the culturing bottle that will load cell places the enclosure of pressure tank.
Here, culturing bottle is the carrying and the culture apparatus of cell, and its structure can be as shown in Figure 3.
The enclosure of pressure tank is the place of testing, thereby before test, need culturing bottle be placed enclosure, to begin test.Certainly, before this step, can carry out disinfection to enclosure, be gnotobasis to guarantee enclosure.
Among the present invention, the shell of pressure tank can not be a complete closed, and opening must be set on shell, and in this open outer side one chamber door is set, thereby makes things convenient for putting into and taking out of culturing bottle.The structure of this chamber door can be circular turn switch door, and it is fitted through plastic pad and case surface, to guarantee resistance to air loss.Thereby before this step, also comprise step 400: the turn switch door of opening pressure case, could carry out like this that this step is described to place enclosure with culturing bottle.After this step finishes, before step 402, also can comprise step 401-2: close the turn switch door, and its plastic pad and case surface are fitted, thereby guarantee that air outside can not get into enclosure through the opening that this turn switch door is sealed.
Step 402: open a scavenging air valve and No. two scavenging air valves; Open gas bomb, make gas wherein arrive enclosure, the original gas of enclosure is discharged from No. two scavenging air valves along an air shooter.
Before testing, need the original gas of enclosure (like air, the used gas of test that carries out before) be discharged, thereby this step to be opened scavenging air valve No. two.And after successively opening scavenging air valve and gas bomb, the gas in the gas bomb just can be automatically along scavenging air valve of an air shooter entering, and exports the entering enclosure from it, and original gas is discharged from No. two scavenging air valves in the shell.
Certainly, the discharge of gas needs certain hour, is above-mentioned 13824cm at the volume of this enclosure
3Situation under, the execution time of this step can be more than 1 minute, to guarantee the purity of enclosure gas.
Step 403: after the original gas of enclosure all is discharged to housing exterior, close scavenging air valve No. two.
In this step, the standard that the original gas of judgement enclosure all is discharged to housing exterior can be the execution time of step 402, certainly; Also can utilize special gas detector to detect; For example, gas detector is placed the exit of No. two scavenging air valves 206 (outlet is communicated with extraneous air) of Fig. 2, detect the composition of institute's expellant gas in the shell 2011; If the composition of this gas reaches preassigned; Explain that promptly the original gas of enclosure all is discharged to housing exterior, so just can carry out this step, close scavenging air valve No. two.
Step 404: when manometric registration reaches predetermined pressure, close gas bomb and a scavenging air valve successively.
The implementing precondition of this step is that scavenging air valve and the gas bomb that step 402 is opened still is in opened condition; Thereby along with the input gradually of gas in the gas bomb, the air pressure in the shell is also raising gradually, when manometric registration (being the air pressure of enclosure) reaches predetermined pressure (promptly predetermined test pressure); Should stop gas from the input of gas bomb to enclosure; At this moment, the closing sequence of a gas bomb and a scavenging air valve is to close gas bomb earlier, closes scavenging air valve again No. one; In order to avoid close the leakage that No. a scavenging air valve causes gas earlier, perhaps gas pours in the leakage that causes a scavenging air valve.
Step 405: when the test duration arrives the scheduled time, open No. two scavenging air valve, the gas of enclosure is discharged from No. two scavenging air valves.
The test duration here is scheduled to, because the shell volume among the present invention will be far longer than the volume of the culturing bottle that prior art provides, thereby should the test duration changeable.
Test duration arrives the end that the scheduled time means test; Should open No. two scavenging air valve this moment once more, so that the gas of enclosure is discharged, if the gas of enclosure is nontoxic; Then can directly be discharged in the atmosphere; If the gas of enclosure is toxic gases such as carbon monoxide, then can it be entered special gas-filtering device or gas concentration unit, in order to avoid contaminate environment.
Step 406: take out culturing bottle from enclosure, the cell of its loading is analyzed.
After carrying out this step, can analyze and research to the cell in the process culturing bottle of pressure test, thereby confirm the influence of predetermined test pressure cell growth.
In order to make probe temperature remain the human body temperature at cell place; This test macro can also be provided with thermostat container; Thereby after above-mentioned step 401, before step 402, can also comprise step 401-1: pressure tank is placed in the thermostat container; And making the thermostat container temperature inside remain preset steady temperature, this preset steady temperature is the temperature (can be about 37 ℃ like somatocyte) of the position of human body at the cell place in the culturing bottle.
Among the present invention; As shown in Figure 2; Shell 2011 inside of pressure tank 201 are divided into superposed culturing room and are positioned at the damp camera of bottom by horizontally disposed internal partition 2013, and have more than one through hole 20131 on the internal partition 2013 so that damp camera and culturing room communicate; Then
Step 401 is: the culturing bottle that will load cell places in the inner culturing room of the shell 2011 of pressure tank 201 (on the upper surface of internal partition 20131, and maintenance through hole 20131 is unimpeded);
In addition; Step 401 can also comprise: the absorbent cotton that will be soaked with sterile distilled water places (top of the lower surface in the shell 2011) in the damp camera; So that shell 2011 humidity inside reach predetermined moisture (identical with the humidity of the position of human body at cell in culturing bottle place, or predetermined testing humidity).
Among the present invention, the gas in the gas bomb can have multiple choices, for example; When the influencing of test low-oxygen environment overdraft cell growth; This gas can be air that contains minor amounts of oxygen (volume ratio is lower than 21%) or the nitrogen that is mingled with minor amounts of oxygen, and when the influencing of test oxygen-free environment overdraft cell growth, this gas can be nitrogen; When the influencing of test CO gas cell growth; Gas in the gas bomb can be carbon monoxide, and under the situation that does not have the gas composition restriction, gas wherein can be and contains the air that volume ratio is 5% carbonic acid gas.
In addition, the present invention also can be used for testing drug pair cell compressive resistance (high pressure) Study on Mechanism, therefore; The medicine of predetermined concentration can also be set in the culturing bottle; Make cell in culturing bottle, carry out contact (as contacting as media through cell culture fluid etc.) of predetermined way, behind EOT, just can analyze and research by pair cell with medicine; Cell situation when medicine not being set is compared, thereby judges the mechanism of action and the usefulness of medicine.
This shows that the present invention has the following advantages:
(1) among the present invention; Culturing bottle is positioned at the shell of pressure tank; Gas bomb links to each other with the inlet of the scavenging air valve that air inlet port was connected of shell through an air shooter; Can when a scavenging air valve is in opened condition, gas be provided,, and make the air pressure of enclosure reach predetermined test pressure with the gas of displacement enclosure to enclosure; The gas of enclosure can be discharged through No. two scavenging air valves that venting hole connected, and the air pressure of enclosure (being the pressure that cell bore in the culturing bottle) can accurately be read by tensimeter.Like this; The space of the enclosure among the present invention is more much bigger than the volume of the culturing bottle about 50ml in the prior art; Even cell is through long respiration; The gas composition of enclosure is taken place than about-face, thereby test result of the present invention can accurately reflect the influence of long-time internal pressure factor for the cell growth.The present invention utilizes tensimeter, can accurately obtain the air pressure in the pressure tank shell, coarse with respect to prior art, and the present invention is accurately for the setting of test pressure, thereby test result also is accurately convictive.
(2) the present invention is arranged at the inside of thermostat container with pressure tank, through regulating the thermostat container temperature inside, makes its temperature that remains position in the human body of cell place, thereby has improved accuracy of the present invention.
(3) the present invention can provide various experimental situation (comprising pressure environment and atmosphere surrounding) through the mode that changes the gaseous constituent in the gas bomb; Cell category to be tested is various; For example can be retinal ganglial cells, tm cells, cardiovascular endotheliocyte etc.; Thereby can carry out the test under the kinds of experiments environment to various kinds of cell; Produce good medical research effect and social benefit, significant to the research and the prevention of blindness of glaucoma, hypertension, heart trouble, cardiovascular disorder.
The above is merely preferred embodiment of the present invention, and is in order to restriction the present invention, not all within spirit of the present invention and principle, any modification of being done, is equal to replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (10)
1. the system of the influence of a test pressure cell growth; It is characterized in that this system comprises: culturing bottle, the gas bomb that gas is housed, tensimeter, a scavenging air valve, No. two scavenging air valves, an air shooter and No. two air shooters of pressure tank, the said cell of loading; Wherein, said pressure tank comprises shell;
Said culturing bottle is positioned at said enclosure;
Have on the said shell and make air inlet port, venting hole and the pressure detection hole that is interlinked to the outside in it;
The outlet of a said scavenging air valve connects said air inlet port, and it enters the mouth through a said said gas bomb of gas delivery pipe coupling;
The inlet of said No. two scavenging air valves connects said venting hole;
One end of said No. two air shooters links to each other with said tensimeter, and the other end communicates with said enclosure through said pressure detection hole.
2. system according to claim 1 is characterized in that, also comprises the thermostat container of the steady temperature that inner maintenance is preset;
Said pressure tank is positioned at the inside of said thermostat container.
3. system according to claim 1 is characterized in that said pressure tank also comprises internal partition, and its level is positioned at the predetermined height of the top, bottom surface of said shell, and said enclosure is divided into superposed culturing room and the damp camera that is positioned at the bottom;
Have more than one through hole on the said internal partition, said damp camera and said culturing room are communicated;
Said culturing bottle is positioned at said culturing room;
This system also comprises the absorbent cotton that is soaked with sterile distilled water, and it is positioned at said damp camera.
4. system according to claim 1 is characterized in that, said pressure tank also comprises circular turn switch door, and it is fitted through plastic pad and said case surface.
5. system according to claim 1 is characterized in that, also comprises the gas concentration unit that links to each other with the outlet of said No. two scavenging air valves.
6. the method for the influence of a test pressure cell growth, this method is based on the described system of claim 1; It is characterized in that this method comprises:
Step 1: the culturing bottle that will load said cell places the enclosure of said pressure tank;
Step 2: open a said scavenging air valve and said No. two scavenging air valves; Open said gas bomb, make gas wherein arrive said enclosure, the original gas of said enclosure is discharged from said No. two scavenging air valves along a said air shooter;
Step 3: after the original gas of said enclosure all is discharged to said housing exterior, close said No. two scavenging air valves;
Step 4: when said manometric registration reaches predetermined pressure, close a said gas bomb and a said scavenging air valve successively;
Step 5: when the test duration arrives the scheduled time, open said No. two scavenging air valves, the gas of said enclosure is discharged from said No. two scavenging air valves;
Step 6: take out said culturing bottle from said enclosure, the said cell of its loading is analyzed.
7. method according to claim 6 is characterized in that, after said step 1, before said step 2, also comprises step 1-1: said pressure tank is placed in the thermostat container, and make said thermostat container temperature inside remain preset steady temperature.
8. method according to claim 6; It is characterized in that; The enclosure of said pressure tank is divided into superposed culturing room and is positioned at the damp camera of bottom by internal partition, and has more than one through hole on the said internal partition so that said damp camera and said culturing room communicate; Then
Said step 1 is: the culturing bottle that will load said cell places in the culturing room of enclosure of said pressure tank;
Said step 1 also comprises: the absorbent cotton that will be soaked with sterile distilled water places in the said damp camera, so that the humidity of said enclosure reaches predetermined moisture.
9. method according to claim 6 is characterized in that,
Before said step 1, also comprise step 0: the turn switch door of opening said pressure tank;
After said step 1, before said step 2, also comprise step 1-2: close said turn switch door, and its plastic pad and said case surface are fitted.
10. method according to claim 6 is characterized in that, the gas in the said gas bomb is: nitrogen; Or, carbon monoxide; Or, contain volume ratio and be the air of 5% carbonic acid gas.
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| CN201110283487A CN102329724A (en) | 2011-09-22 | 2011-09-22 | System and method for testing influence of pressure on cell growth |
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| CN201110283487A CN102329724A (en) | 2011-09-22 | 2011-09-22 | System and method for testing influence of pressure on cell growth |
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Application publication date: 20120125 |