CN103333800A - Dynamic-static positive-negative pressure loading experiment system and method for in-vitro cells - Google Patents
Dynamic-static positive-negative pressure loading experiment system and method for in-vitro cells Download PDFInfo
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- CN103333800A CN103333800A CN2013102547494A CN201310254749A CN103333800A CN 103333800 A CN103333800 A CN 103333800A CN 2013102547494 A CN2013102547494 A CN 2013102547494A CN 201310254749 A CN201310254749 A CN 201310254749A CN 103333800 A CN103333800 A CN 103333800A
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Abstract
The invention relates to a dynamic-static positive-negative pressure loading experiment system and method for in-vitro cells. The system comprises a gas-source gas storage bottle, a pressure reduction valve, an electromagnetic gas valve, a filter, a diverter, a high-voltage buffering container, a normal-voltage buffering container, a high-voltage container, a low-voltage container, a vacuum pump, a compression pump, a pressure culture box, a digital display temperature transducer, a digital display pressure meter strap transducer, a data acquisition system, a driving control system, a heater and a water bath tank, wherein the gas-source gas storage bottle is connected with the pressure culture box through the pressure reduction valve, the electromagnetic gas valve, the filter and the pressure container; the pressure culture box is connected with the vacuum pump and the compression pump through the pressure container; and the pressure culture box is also connected with the data acquisition system and the driving control system through a pressure sensor and a temperature sensor. The dynamic and static positive-negative pressure loading experiment system can simulate an in-vivo mechanical environment of multiple cells in vitro, in particular a pressure environment of a temporal jaw joint cavity and supply a research platform to in-vitro research on a biomechanical response of in-vivo cells.
Description
Technical field
The present invention relates to the moving static quadrature reactance of a kind of cell in vitro-negative pressure loading experiment system and method thereof, belong to cell biological mechanics and organizational engineering field.
Background technology
The cell biological mechanics is an interactional science of research cell and mechanical factor, is the important integral part of organizational engineering.Organism is under the effect of various mechanical forces constantly, and also often be under pressure in body tissue and cell, multi-form effect such as shearing force, distraction force, force of compression, constitute a mechanical system under the different mechanical functions, in the multiple activity of cell such as propagation, differentiation, adhesion, migration, apoptosis, play an important role.But it is single effect or joint effect that numerous mechanics factors is difficult to distinguish in vivo.Therefore, the effect in activities such as the growth of cell, differentiation has directive significance to the biomechanics effect of studying single mechanics factor for clear and definite mechanical stimulation.At present, the development of cell in vitro Experiments of Machanics deceleration loading device has become the main method of research cell biological mechanics.Pressure is the common mechanics form of cell in vivo, and is the most common in the joint.Temporomandibular joint is bearing in moving process from malleation and is changing with negative pressure and along with the different mode of motion of lower jaw, and therefore, the pressure environment that can simulate temporomandibular joint is significant with regeneration in the cartilage reconstruction for research condylar cartilage cell.
Development along with tissue engineering technique, stem cell is with its vigorous propagation and differentiation potential, for tissue transplantation with reproduce better method is provided, but its maximum defective is the cartilage cell epimatrix mechanical property deficiency that generates, and stem cell can be divided into the more cartilaginous tissue of horn of plenty of matrix components under suitable pressure effect.Therefore, design the moving static quadrature reactance of a kind of multi-functional cell in vitro-negative pressure loading experiment system and study the especially biomechanical response under the effect of condylar cartilage cyto-mechanics of various kinds of cell for us, further the mechanical signal transmission mechanism of the form of announcement cell cell under different pressure action conditions, propagation, differentiation, apoptosis is set up the in vitro study model to organizational engineering.
At present, the pressure-loaded experimental installation of foreign study cell is that cell pressure-loaded system with Yousefian development is representative.And at home, this area has been developed dissimilar cell pressure-loaded experimental installations, and still, these experimental installations are the limitation of various degrees also.Specifically comprise: (1) though existing experimental installation can load static pressure and dynamic pressure, also not to dynamic negative-pressure and dynamically just-and negative pressure simulates, and is difficult to the satisfied needs that temporomandibular joint is studied.(2) existing experimental installation can't be regulated and control the pressure-loaded environment of cell, is difficult to satisfy to the long pressure-loaded experiment of cell.(3) pattern of pressure of existing experimental installation is more single, and the narrow range of pressure-loaded is difficult to satisfy the research of different pressures environment cell in the body.
Summary of the invention
A kind of cell in vitro that the objective of the invention is to develop moves static quadrature reactance-negative pressure loading experiment system and a kind of method of cell pressure-loaded is provided, be characterized at the external pressure that can load various ways under the culture condition that cell is being specified, comprise (static malleation, static negative pressure, dynamically malleation, dynamic negative-pressure and dynamically just-negative pressure), and can control and regulate size, frequency, action time and the pressure binding mode of on-load pressure.Experimental system can be under the prerequisite that does not influence pressure the gas in the discharge section experimental system and open the tonifying Qi of air inlet magnetic valve by the program of setting regularly, change gas in the whole experimental system according to the time of setting, keep the CO of culture environment
2Dividing potential drop and O
2Dividing potential drop is constant, thereby keeps pH value and the HCO of cell culture fluid
3 -Steady concentration.Under different pressure binding modes, keep-up pressure by two kinds of type of heating and cultivate the constant of the temperature inside the box, by the keep-up pressure three-dimensional environment of the cell cultures in the incubator of these methods.That this pilot system has is simple to operate, the advantage of stable performance, precise control, and mechanical environment that can be in in-vitro simulated various kinds of cell body is set up in vitro study at the platform of somatic biomechanical response.
The used cell pressure-loaded experimental system of the present invention is characterized in can simulating at somatic mechanical environment, comprises static malleation, static negative pressure, dynamic malleation, dynamic negative-pressure and dynamically just-negative pressure.The principle of work of system is: culture dish or the culture plate of culturing cell are put into incubator, open the bomb switch, gas passes through reducing valve from the source of the gas bomb, electromagnetic gas valve and strainer enter the four-way splitter, enter the normal pressure buffer container again, the gas of normal pressure buffer container pumps into high-pressure buffer container and low pressure vessel by compression pump, pass through the five-way splitter by the control of high pressure electromagnetic gas valve and low voltage electromagnetic air valve respectively again and enter into the pressure incubator, gas in the incubator enters the normal pressure buffer container by five-way splitter and normal pressure electromagnetic gas valve again, forms the closed cycle of a gas.Gas in the incubator also can be discharged by the exhaust electromagnetic gas valve, to finish ventilation.At this moment, according to pressure binding mode, pressure size, frequency, the action time set, the control mainboard is by driving control system control compression pump and vacuum pump work, for corresponding static malleation, static negative pressure, dynamic malleation, dynamic negative-pressure and dynamically just-negative pressure are provided in the incubator, pressure and temperature in pressure incubator and each pressure buffer container all can be finished collection by data collecting system by each pressure transmitter and temperature sensor simultaneously, but control mainboard indicating system each several part pressure and temperature parameter, and regulate according to setting program.In the experimentation, can pass through the situation of the transparent toughened glass observation of cell cultivation of pressure incubator hatch door, after the working hour was set in arrival, the control mainboard was according to the parameter end pressure-loaded of setting.Open exhaust solenoid valve, the gas of emptying incubator is opened the incubator hatch door.
The purpose of invention is realized by following technical measures:
This device comprises: contain 5%CO
2Source of the gas, pressure incubator, pressure controlling system, temperature adjusting system, data collecting system and the driving control system of mixing air.The source of the gas bomb links to each other with the pressure incubator by first reducing valve, electromagnetic gas valve, strainer, four-way splitter, normal pressure buffer container, high-pressure buffer container, low pressure vessel, vacuum pump, compression pump; The other end of pressure incubator links to each other with band sourdine exhaust electromagnetic gas valve; Band sourdine exhaust electromagnetic gas valve links to each other with driving control system and is presented on the display interface; Pressure incubator one end and digital display manometer band transmitter, the first digital display temperature transmitter, the other end is connected with data collecting system; High-pressure buffer container, low pressure vessel are connected with pressure indicator; Normal pressure buffer container one end is connected with pressure unit, and the other end is connected with data collecting system; High-pressure buffer container one end is connected with the pressure incubator by electromagnetic gas valve, and the other end is connected to driving control system by splitter and compression pump; Low pressure vessel one end is connected with the pressure incubator by electromagnetic gas valve, and the other end is connected to driving control system by splitter and vacuum pump; Normal pressure buffer container, high-pressure buffer container, low pressure vessel are inserted in the water bath; Water bath one end connects well heater, and the other end connects water level switch, the digital display temperature transmitter is connected with data collecting system; Pressure controlling system, temperature adjusting system, data collecting system and driving control system are connected and are presented on the control panel with the control mainboard respectively.
Pressure is cultivated in the casing and is established three layers of culture dish support, support is the hollow out type, can hold experiment orifice plate and culture dish, and make the interior gas of incubator easily with the gaseous interchange in the culture dish, the inlet mouth of pressure incubator is arranged on the side of incubator, and baffle plate is set prevents because erratic flowing appears in gas in air inlet and the incubator that causes of giving vent to anger frequently, thereby avoid the pressure in the incubator inhomogeneous.
The incubator hatch door is designed to 4 screws and fixes, and the hatch door mouth seals with sealing-ring, and there is circular viewing window in hatch door central authorities, are high-strength armoured-glass, can realize the omnidistance transparent observing of cell pressurization.
Cell in vitro of the present invention moves static quadrature reactance-negative pressure loading method and may further comprise the steps:
(1) subject cell is put into the pressure incubator together with culture dish, close the incubator hatch door, the seal-off pressure culture chamber;
(2) open the bomb air valve, gas by reducing valve, strainer, normal pressure buffer container enters into high-pressure buffer container and low pressure vessel by compression pump and vacuum pump respectively, enter into incubator by solenoid control again, pressure in the incubator enters the normal pressure buffer container by magnetic valve, forms the closed cycle of a gas and is full of 95% air and 5%CO
2Mixed gas, water bath and fixedly heating unit make the temperature in the incubator reach 36 ± 2 ℃;
(3) set static pressure pattern, action time, compression pump/vacuum pump pattern or dynamic pressure pattern, action time, compression pump and vacuum pump pattern, frequency, regulate the incubator internal pressure by driving control system driving power system and magnetic valve;
(4) pressure incubator internal pressure is communicated to computer main board by pressure transmitter, by the program that configures regulate and the control incubator in pressure;
(5) for the loading regime that requires static pressure, the control mainboard passes through drive compression pump/vacuum pump, makes the pressure in the incubator reach set(ting)value, regulates the variation of pressure simultaneously according to the program of setting, and controls the constant of incubator internal pressure.
(6) for requiring the dynamic pressure loading regime, the control mainboard is by drive compression pump and vacuum pump, and make compression pump and vacuum pump coordination operation, regulate the variation of pressure tank internal gas pressure, make the incubator internal pressure according to frequency and the size variation set, and by the monitoring of pressure sensing transmitter, reach requirement of experiment.
Source of the gas is 5%CO
2With 95% Air mixing gas, utilize medical air filter cleaning gas, guarantee the gas gnotobasis; The pressure incubator selects for use pollution-free, avirulent medical acid-resistant stainless steel materials processing to form, and pressure incubator inwall designs for circular arc, and cleaning and disinfection is convenient at no dead angle.
Type of heating is that Water Tank with Temp.-controlled and assisted heating device are united heating, when being set at static pressure because gas do not flow in incubator, therefore control mainboard control fixedly heating unit be the incubator direct heating.When setting pressure is dynamic pressure, because gas flows rapidly between pressure incubator, normal pressure buffer container, high-pressure buffer container and low pressure vessel, have heat and scatter and disappear, the control mainboard can control Water Tank with Temp.-controlled and fixedly heating unit heat jointly, and keep homo(io)thermism in the incubator.
Data collecting system comprises pressure sensing transmitter, temperature sensing transmitter, and is connected with the control mainboard.
The program that the control mainboard writes is the core of regulation and control pressure change, and its function is as follows:
Drive compression pump and vacuum pump produce air pressure, make and produce certain pressure in high-pressure buffer container and the low pressure vessel, high-pressure buffer container, low pressure vessel circulate mutually with incubator, and to producing the dynamic or static pressure of certain frequency, size in the pressure incubator, monitor the running status of compression pump and vacuum pump simultaneously, make the pressure culture systems stable by driving control system is regulated.
The solenoid control function: the switch by control mainboard control magnetic valve is regulated air inlet and exhaust, thus by the control magnetic valve switching time adjustments of gas flow accurately regulate the incubator internal pressure.
Data processing and Presentation Function: according to the frequency of setting, pressure, working hour, moving/the static pressure pattern, control mainboard drive compression pump and vacuum pump work, by pressure buffer container the pressure in the incubator is regulated, produced a certain size static pressure or the dynamic pressure of certain frequency and size.Simultaneously, by data collecting system the temperature in the pressure incubator, pressure are carried out data gathering, and be presented on the display interface, dynamically demonstrate the process of pressure change simultaneously with wave, can monitor in real time the temperature in the incubator, pressure.
The present invention has following advantage:
(1) type of heating of the present invention be Water Tank with Temp.-controlled and fixedly heating unit unite heating, under the static pressure pattern, gas remains static in incubator, fixedly heating unit work and keep the incubator homo(io)thermism at 37 ℃.Under the dynamic pressure pattern, gas flows in normal pressure buffer container, high-pressure buffer appearance, high pressure vessel and low pressure vessel and pressure incubator fast, Water Tank with Temp.-controlled is the pressurized vessel heating that is immersed in the water-bath, guarantee that the gas temperature in the total system keeps constant temperature, and the scattering and disappearing of gas heat in the make-up system pipeline, guarantee the stable of whole cell culture environment constant temperature by this type of heating, guarantee cell bio-activity, reach cell constant temperature culture environment preferably.Simultaneously, under the dynamic pressure pattern, gas through heat production after the Fast Compression, can cause the temperature in the cell culture incubator sharply to change, and gas can keep cultivating the constant of the temperature inside the box after exchanging between pressure incubator and the pressure buffer container in the pressure incubator.
(2) in the present invention, what the pressure-driven system adopted is compression pump and vacuum pump, in a big way malleation and negative pressure can be provided, pressure buffer container and pressure incubator all adopt high voltage bearing stainless steel to make simultaneously, therefore can carry out that pressure range is wide, pressure pattern is many to the cell cyto-mechanics loading of (static negative pressure, static malleation, dynamic negative-pressure, dynamically malleation and dynamically just-negative pressure), different pressure environments is provided, can be applicable to the various kinds of cell mechanics study.
(3) in the present invention, because the pressure incubator is closed environment, can change the proportion of composing of gas under the long pressurized state of cell, the mainboard Controlling System is opened the tonifying Qi of air inlet magnetic valve according to the program of setting in according to the gas in the program timing discharge section experimental system of setting, and change the gas in the whole experimental system and do not influence the stability of system in the interior time of setting, keep the CO of culture environment
2Dividing potential drop and O
2Dividing potential drop is constant, thereby keeps pH value and the HCO of cell culture fluid
3 -Steady concentration guarantees the good living environment of cell.
(4) in the present invention, control panel is simple to operation, the pressure, temperature, frequency, action time, the pressure of buffer container, variation track and the pressure waveform curve of pressure incubator internal pressure that can show the pressure incubator in real time, and monitor in real time to the parameters to incubator by data collecting system.
(5) in the present invention, the intellectuality of sound attitude pressure-controlling: under static schema, compression pump or vacuum pump work, after pressure incubator inside reached setting pressure, compression pump or vacuum pump quit work.When the incubator air pressure inside changed, after data collecting system detected and collects pressure data, air inlet magnetic valve or exhaust solenoid valve were opened in the order of control mainboard, regulate the incubator internal gas, keep the incubator pressure inside constant.
Description of drawings
Fig. 1 is the moving static quadrature reactance of cell in vitro-negative pressure loading experimental apparatus
Among the figure: 1, the source of the gas bomb, 2, first reducing valve, 3, electromagnetic gas valve, 4, strainer, 5, the four-way splitter, 6, the normal pressure buffer container, 7, vacuum pump, 8, compression pump, 9, safety valve, 10, high pressure vessel, 11, the first pointer tensimeter, 12, second reducing valve, 13, low pressure vessel, 14, the high-pressure buffer container, 15, the second pointer tensimeter, 16, first pressure unit, 17, the 3rd pointer tensimeter, 18, the low voltage electromagnetic air valve, 19, the 4th pointer tensimeter, 20, second pressure unit, 21, the high pressure electromagnetic gas valve, 22, band sourdine exhaust electromagnetic gas valve, 23, the normal pressure electromagnetic gas valve, 24, the five-way splitter, 25, the pressure incubator, 26, the first digital display temperature transmitter, 27, digital display manometer band transmitter, 28, data collecting system, 29, display interface, 30, operating panel, 31, the control mainboard, 32, switch power supply, 33, driving control system, 34, well heater, 35, the second digital display temperature transmitter, 36, water bath, 37, water level switch.
Fig. 2 is pressure incubator synoptic diagram
Among the figure: 25, pressure incubator, 26, the digital display temperature transmitter, 27, digital display manometer band transmitter, 38, the air inlet air outlet, 39, Tissue Culture Dish, 40, the toughened glass viewing window, 41, the culture dish support, 42, the cell cultures box bracket.
Fig. 3 is the moving static quadrature reactance of cell in vitro-negative pressure loading experiment system display interface
Fig. 4 is the programflow chart of the moving static quadrature reactance of cell in vitro-negative pressure loading experiment system
Embodiment
Below in conjunction with accompanying drawing principle of the present invention and feature are described, institute gives an actual example and just is used for explaining and further specifying the present invention, and non-limiting scope of the present invention.Various nonessential improvement and adjustment that the researchist who is skilled in technique in this area makes technical scheme of the present invention all should be included protection scope of the present invention in.
Enter four-way splitter 5 with reference to the gas in the accompanying drawing 1 source of the gas bomb 1 of the present invention by first reducing valve 2, electromagnetic gas valve 3 and strainer 4.Four-way splitter 5 other three ends are connected with normal pressure buffer container 6, vacuum pump 7, compression pump 8 respectively.Compression pump 8 is connected with high pressure vessel 10, safety valve 9, the first pointer tensimeter 11 are connected with high pressure vessel 10, high pressure vessel 10 is connected with high-pressure buffer container 14 by second reducing valve 12, the pressure that the second pointer tensimeter 15 shows in the high-pressure buffer container 14, the gas in the high-pressure buffer container 14 is connected with five-way splitter 24 by high pressure electromagnetic gas valve 21.One end of low pressure vessel 13 is connected with vacuum pump 7, and the other end of low pressure vessel 13 is connected with an end of low voltage electromagnetic air valve 18, the other end of low voltage electromagnetic air valve 18 be connected with five-way splitter 24.First pressure unit 16, the 3rd pointer tensimeter 17 are sampled to gaseous tension in the low pressure vessel 13 respectively and are shown.One end of normal pressure buffer container 6 is connected with four-way splitter 5, and the other end is connected with normal pressure electromagnetic gas valve 23; The 4th pointer tensimeter 19, second pressure unit 20 show gaseous tension in the normal pressure buffer container 6 respectively and sample.Band sourdine exhaust electromagnetic gas valve 22 is connected with five-way splitter 24; Pressure incubator 25 is connected with five-way splitter 24, and the first digital display temperature transmitter 26, digital display manometer band transmitter 27 show gaseous tension and temperature in the normal pressure buffer container 6 respectively and sample.Water in 34 pairs of water baths 36 of well heater heats, and the water temperature in 35 pairs of water baths 36 of the second digital display temperature transmitter is gathered, and the water level in 37 pairs of water baths 36 of water level switch is gathered.Data collecting system 28 is connected with each data sampling sensor of system, and is connected with control mainboard 31, and switch power supply 32, driving control system 33, display interface 29 and operating panel 30 are connected with control mainboard 31 respectively.
Pressure is cultivated in the casing 25 and is established three layers of culture dish support 41, support is the hollow out type, can hold Tissue Culture Dish 39, and make gas reach setting air pressure with the gaseous interchange in the culture dish easily, the inlet mouth 38 of pressure incubator is arranged on the side of incubator simultaneously, and baffle plate is set prevents from because of air inlet frequently and give vent to anger causing the gas in the incubator erratic flowing to occur, thereby avoid the pressure in the incubator inhomogeneous.
The incubator hatch door is designed to 4 screws and fixes, and the hatch door mouth seals with sealing-ring, and there is circular tempering glass window 40 in hatch door central authorities, can realize the omnidistance transparent observing of cell pressurization.
Cell in vitro of the present invention moves static quadrature reactance-negative pressure loading method and may further comprise the steps:
(1) opens source of the gas bomb air valve, source of the gas gas-holder and pressure are cultivated in the casing be full of 95% air and 5%CO
2Mixed gas;
(2) culture dish is put into the pressure incubator, closed and airtight described pressure incubator;
(3) set pressure pattern, pressure size, frequency and the action time that needs, the control mainboard is controlled magnetic valve by driving circuit, regulates and controls described pressure and cultivate loading pressure;
(4) described pressure is cultivated loading pressure and is conveyed to the control mainboard by pressure transmitter, regulates in the described pressure incubator variation of gas pressure and reaches setting pressure by program;
(5) described pressure is cultivated the temperature inside the box and is sent to the control mainboard by temperature sensor, regulates the variation of variation of temperature and holding temperature by program;
(6) for the loading regime of static pressure, by setting experiment parameter at control panel, comprise the pressure size, just/negative pressure pattern, action time, compression pump/vacuum pump pattern, described control mainboard is by driving control system drive compression pump/vacuum pump, regulate the pressure in the described pressure incubator, digital display temperature transmitter and digital display pressure unit be monitor temperature and pressure respectively, and being sent to the control mainboard by data collecting system, the control mainboard is regulated by setup parameter and is guaranteed that the pressure and temperature in the described pressure incubator is constant;
(7) for the loading regime of dynamic pressure, by setting experiment parameter at described control panel, comprise the pressure size, dynamic malleation/dynamic negative-pressure/dynamically just-the negative pressure pattern, frequency, action time, compression pump+vacuum pump pattern, the control mainboard is by driving control system drive compression pump and vacuum pump, regulate the variation of the pressure in the described pressure incubator, digital display temperature transmitter and digital display pressure unit be monitoring pressure and temperature parameter respectively, and be sent to the control mainboard by data collecting system, adjust the pressure and temperature in the described pressure incubator, and the dynamic pressure that reaches in the described pressure incubator changes.
Claims (5)
1. the moving static quadrature reactance of cell in vitro-negative pressure loading experiment system is characterized in that this system comprises source of the gas bomb (1), first reducing valve (2), electromagnetic gas valve (3), strainer (4), four-way splitter (5), normal pressure buffer container (6), vacuum pump (7), compression pump (8), safety valve (9), high pressure vessel (10), the first pointer tensimeter (11), second reducing valve (12), low pressure vessel (13), high-pressure buffer container (14), the second pointer tensimeter (15), first pressure unit (16), the 3rd pointer tensimeter (17), low voltage electromagnetic air valve (18), the 4th pointer tensimeter (19), second pressure unit (20), high pressure electromagnetic gas valve (21), band sourdine exhaust electromagnetic gas valve (22), normal pressure electromagnetic gas valve (23), five-way splitter (24), pressure incubator (25), the first digital display temperature transmitter (26), digital display manometer band transmitter (27), data collecting system (28), display interface (29), operating panel (30), control mainboard (31), switch power supply (32), driving control system (33), well heater (34), the second digital display temperature transmitter (35), water bath (36), water level switch (37).
Wherein, gas in the described source of the gas bomb (1) is by described first reducing valve (2), electromagnetic gas valve (3) and strainer (4) enter an end (5) of described four-way splitter, other three ends of described four-way splitter (5) respectively with described normal pressure buffer container (6), vacuum pump (7), compression pump (8) is connected, described compression pump (8), safety valve (9), the first pointer tensimeter (11) is connected with described high pressure vessel (10) respectively, described high pressure vessel (10) is connected with described high-pressure buffer container (14) by described second reducing valve (12), the described second pointer tensimeter (15) shows the pressure in the described high-pressure buffer container (14), gas in the described high-pressure buffer container (14) is connected with described five-way splitter (24) by described high pressure electromagnetic gas valve (21), one end of described low pressure vessel (13) is connected with described vacuum pump (7), the other end of described low pressure vessel (13) is connected with an end of described low voltage electromagnetic air valve (18), the other end of described low voltage electromagnetic air valve (18) be connected with described five-way splitter (24), described first pressure unit (16), the 3rd pointer tensimeter (17) is sampled to gaseous tension in the described low pressure vessel (13) respectively and is shown, one end of described normal pressure buffer container (6) is connected with described four-way splitter (5), and the other end of described normal pressure buffer container (6) is connected with described normal pressure electromagnetic gas valve (23); Described the 4th pointer tensimeter (19), second pressure unit (20) show to gaseous tension in the described normal pressure buffer container (6) respectively and sample that described band sourdine exhaust electromagnetic gas valve (22) is connected with described five-way splitter (24); Described pressure incubator (25) is connected with described five-way splitter (24), the described first digital display temperature transmitter (26), digital display manometer band transmitter (27) shows gaseous tension in the described normal pressure buffer container (6) and temperature respectively and samples, described well heater (34) heats the water in the described water bath (36), the described second digital display temperature transmitter (35) is gathered the water temperature in the described water bath (36), described water level switch (37) is gathered the water level in the described water bath (36), described data collecting system (28) and the first digital display temperature transmitter (26), digital display manometer band transmitter (27), the second digital display temperature transmitter (35) is connected, and described data collecting system (28) is connected with described control mainboard (31), described switch power supply (32), driving control system (33), display interface (29) is connected with described control mainboard (31) respectively with operating panel (30).
2. cell in vitro moves static quadrature reactance-negative pressure loading experiment system according to claim 1, it is characterized in that, establish three layers of culture dish support in the described pressure incubator (25), support is hollow out shape, can hold experiment with orifice plate and culture dish, make gas easily with the gaseous interchange in the culture dish and reach setting air pressure.The hatch door central authorities of described pressure incubator (25) arrange window, are transparent toughened glass, can carry out visible observation to the cell whole process of pressurizeing.
3. the moving static quadrature reactance of cell in vitro-negative pressure loading experiment system according to claim 1 is characterized in that described high-pressure buffer container (14), high pressure vessel (10), low pressure vessel (13), normal pressure buffer container (6) and vacuum pump (7), compression pump (8) formed the pressure-driven system.
4. cell in vitro moves static quadrature reactance-negative pressure loading experiment system according to claim 1, it is characterized in that, by place the high-pressure buffer container (14) of water bath (36) with well heater (34) heating, low pressure vessel (13), normal pressure buffer container (6), gas in the heating system, and by being placed on the described pressure incubator of fixedly heating unit direct heating (25) of pressure incubator (25) below, thereby control and regulate temperature in the pressure incubator by control mainboard (31), the described first digital display temperature transmitter (26) is sampled to the temperature in the incubator and is shown.
5. as the pressure-loaded method of the moving static quadrature reactance of cell in vitro as described in the claim 1-4-negative pressure loading experiment system, it is characterized in that said method comprising the steps of:
(1) opens source of the gas bomb air valve, source of the gas gas-holder and pressure are cultivated in the casing be full of 95% air and 5%CO
2Mixed gas;
(2) culture dish is put into the pressure incubator, closed and airtight described pressure incubator;
(3) set pressure pattern, pressure size, frequency and the action time that needs, the control mainboard is controlled magnetic valve by driving circuit, regulates and controls described pressure and cultivate loading pressure;
(4) described pressure is cultivated loading pressure and is conveyed to the control mainboard by pressure transmitter, regulates in the described pressure incubator variation of gas pressure and reaches setting pressure by program;
(5) described pressure is cultivated the temperature inside the box and is sent to the control mainboard by temperature sensor, regulates the variation of variation of temperature and holding temperature by program;
(6) for the loading regime of static pressure, by setting experiment parameter at control panel, comprise the pressure size, just/negative pressure pattern, action time, compression pump/vacuum pump pattern, described control mainboard is by driving control system drive compression pump/vacuum pump, regulate the pressure in the described pressure incubator, digital display temperature transmitter and digital display pressure unit be monitor temperature and pressure respectively, and being sent to the control mainboard by data collecting system, the control mainboard is regulated by setup parameter and is guaranteed that the pressure and temperature in the described pressure incubator is constant;
(7) for the loading regime of dynamic pressure, by setting experiment parameter at described control panel, comprise the pressure size, dynamic malleation/dynamic negative-pressure/dynamically just-the negative pressure pattern, frequency, action time, compression pump+vacuum pump pattern, the control mainboard is by driving control system drive compression pump and vacuum pump, regulate the variation of the pressure in the described pressure incubator, digital display temperature transmitter and digital display pressure unit be monitoring pressure and temperature parameter respectively, and be sent to the control mainboard by data collecting system, adjust the pressure and temperature in the described pressure incubator, and the dynamic pressure that reaches in the described pressure incubator changes.
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| CN113801791A (en) * | 2021-03-26 | 2021-12-17 | 中国海洋大学 | Multichannel uniform tensile stress in-vitro cell culture device and working method |
| CN115369037A (en) * | 2022-10-24 | 2022-11-22 | 中国中医科学院望京医院(中国中医科学院骨伤科研究所) | Full circulation device, method and loading system for spinal culture solution |
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| CN109581230A (en) * | 2018-11-27 | 2019-04-05 | 上海理工大学 | Soft-package battery outer surface constant pressure that compressed gas is adjustable pre-tightens test device and method |
| CN113801791A (en) * | 2021-03-26 | 2021-12-17 | 中国海洋大学 | Multichannel uniform tensile stress in-vitro cell culture device and working method |
| CN113801791B (en) * | 2021-03-26 | 2022-08-02 | 中国海洋大学 | Multichannel uniform tensile stress in-vitro cell culture device and working method |
| WO2023232099A1 (en) * | 2022-06-02 | 2023-12-07 | 北京中赢谷投资管理有限公司 | Method for obtaining vesicles |
| CN115369037A (en) * | 2022-10-24 | 2022-11-22 | 中国中医科学院望京医院(中国中医科学院骨伤科研究所) | Full circulation device, method and loading system for spinal culture solution |
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