CN103361270B - Multi-scenarios method In vitro cell experiment device and method - Google Patents
Multi-scenarios method In vitro cell experiment device and method Download PDFInfo
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- CN103361270B CN103361270B CN201310304190.1A CN201310304190A CN103361270B CN 103361270 B CN103361270 B CN 103361270B CN 201310304190 A CN201310304190 A CN 201310304190A CN 103361270 B CN103361270 B CN 103361270B
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Abstract
The invention discloses a kind of multi-scenarios method In vitro cell experiment device and method, described device comprises blind flange and the multifunctional sample pallet that container, surface that insulating interlayer and magnetic stirring apparatus are housed are provided with multiple perforate; The top and bottom of described container are respectively arranged with filler and dewatering outlet, described dewatering outlet place is provided with ball valve, described magnetic stirring apparatus is in order to balanced bath temperature, the top that described blind flange is connected to container is formed can the structure of opened/closed container, described sample pallet can place electromagnetic field generator with porous cell culture plate and/or single porous cell culture plate, is placed in container when testing; Described method is placed on sample pallet by the electromagnetic field generator with porous cell culture plate, then puts into container, then control atmosphere needed for cell cultures, temperature and humidity condition realizes.Multi-scenarios method In vitro cell experiment apparatus structure of the present invention is simple, with low cost, easy to operate, diverse in function, and can meet and carry out organizing experiment more simultaneously.
Description
Technical field
The present invention relates to a kind of cell culture apparatus and method, especially a kind of multi-scenarios method In vitro cell experiment device and method, belongs to medical instruments field.
Background technology
Natural somatocyte is in complicated body microenvironment, and the effect of the stress-strain of natural body, bio-electric field, biological magnetic field, affects the structure of body tissue growth, the development of morphology and function.From nineteen fifty-seven Fukada and Yasuda, the discovery of the macroforce electrical property of bone and the stress of Bassett discovery are subsequently promoted that the reason of bone remoulding may be the current potential that stress produces in bone, have stimulated cell response, induction of the physiological change of cell.
Biomechanics has in clinical application: the limbs lengthening art of orthopaedics, the pressurization ligamentopexis of fracture, cranium Jaw distraction art and skin soft tissue stretching art etc.The action of a magnetic field aspect, animal experiment is had to find: under the action of a magnetic field, the bone trabecula number of experimental group and thickness all increase, the maximum tensile strength of femur and the maximum compressive strength of centrum all reach normal, and the bone surface such as human spine, femur and inside will produce time dependent induced current.By electro-magnetic biological effect, affect bone cell metabolism, improve bone density, improve bone mass, reach the object of prevention and therapy osteoporosis and fracture.Bioelectrical effect is the moulding of bone and rebuild important role, and union of fracture is the methods for the treatment of that orthopaedics is conventional to adopt electricity irritation to promote.The charge accumulated formed at fracture applying negative voltage is conducive to union of fracture.A large amount of experimentation on animalies also proves, continuous direct current, and pulse direct current, lasting electrostatic field and pulse electromagnetic field all can promote growth and the reparation of bone, and widespread use in clinical treatment.
Utilize the clinical treatment of Biological Strength, electricity, magnetic field effect to be widely used, but its Molecular Biology Mechanism is set forth not yet completely.To the in vitro study of active somatic cell under power, electricity, magnetic field environment to inquire in it the Molecular Biology Mechanism significant.The research of In vitro cell experiment is except building the power of setting, electricity, magnetic field, and its experimental situation must adapt with the cell culture environment of routine, otherwise can not accurate simulation living organism internal milieu, causes its result of study to lack scientific basis.
At present, the field of force is applied to cell, experimental installation that magnetic field and electric field are studied has four-point bending cell augmenter, pore pressurizing cell cultivation instrument, cell in vitro loading device with static magnetic field and cell electrophoresis apparatus etc., however these equipment and instruments exist that application function is single, complex operation, price are high, can not independently carry out cell cultures, be difficult to realize many group experiments carry out simultaneously, be unfavorable for studying biomaterial and cell response this type of need the multiple not convenient factors such as the experimental study of a large amount of sample.
Summary of the invention
The object of the invention is the defect in order to solve above-mentioned prior art, providing that a kind of structure is simple, with low cost, easy to operate, diverse in function, and the multi-scenarios method In vitro cell experiment device simultaneously carrying out many group experiments can be met.
Another object of the present invention is to a kind of multi-scenarios method In vitro cell experiment method that said apparatus is provided.
Object of the present invention can reach by taking following technical scheme:
Multi-scenarios method In vitro cell experiment device, is characterized in that: comprise can constant control water temperature container, to match with container and surface be provided with multiple perforate blind flange, the uniform magnetic stirring apparatus of distribution of water temperature can be realized and the sample pallet of the electromagnetic field generator that can place with porous cell culture plate and/or single porous cell culture plate; The top and bottom of described container are respectively arranged with filler and dewatering outlet, and described dewatering outlet place is provided with ball valve, and the top that described blind flange is connected to container is formed can the structure of opened/closed container, and described magnetic stirring apparatus is arranged on the bottom of container; Described sample pallet is placed in container when testing.
As a kind of preferred version, described container lower end is also provided with heating tube and the first thermometer hole for placing the first thermometer, and described heating tube is connected with outside temperature regulating device respectively with the first thermometer, realizes constant control water temperature.
As a kind of preferred version, the perforate that described blind flange surface is arranged comprise air inlet port, production well, line terminal, for place barometric weather gage hole, for place the second thermometer the second thermometer hole and for placing hygroscopic wet bulb thermometer hole.
As a kind of preferred version, described electromagnetic field generator comprises one group of parallel Helmholtz coil, one group of parallel-plate electrode, dull and stereotyped pallet and power supply, and described parallel-plate electrode is placed on dull and stereotyped pallet, is provided with terminal stud bottom described power supply; When electromagnetic field generator is placed in container, the wiring of described terminal stud is connected with external power source control device by line terminal.
As a kind of preferred version, described sample pallet comprises Multilayer Samples platform and at least three for supporting the thread rack of sample table, described every layer of sample table is the irregular roundness shape cropping part arc length, its surface is provided with at least three bracket holes matched with thread rack, and described thread rack is through being regulated by hex-nut after bracket holes and the height of fixed sample platform.
As a kind of preferred version, the surface of described every layer of sample table is also provided with multiple boring for placing centrifuge tube.
As a kind of preferred version, described container is the cylindrical straight tubular that insulating interlayer is housed, and conveniently sample pallet is put into or proposed, and improve the homogeneity of internal temperature, it adopts stainless material to make, and can stablize pressurize.
As a kind of preferred version, between described blind flange and the top of container, be provided with rubber cradle, to improve the sealing effectiveness of container.
Another object of the present invention can reach by taking following technical scheme:
Multi-scenarios method In vitro cell experiment method, is characterized in that comprising the following steps:
1), after being sterilized by container sterilization, in container, a certain amount of deionized water is added to keep the humidity environment of cell cultures:
2) the porous cell culture plate being placed with cell sample is placed between the parallel-plate electrode of electromagnetic field generator;
3) electromagnetic field generator is placed on sample pallet, then sample pallet is placed in container, the wiring of terminal stud is connected with external power source control device by line terminal, by blind flange unopened container;
4) by air inlet port and band rate of flow meter containing 5%CO
2medical mixed gas gas-holder be connected, then adjust pore, monitored by weather gage, obtain stable air pressure;
5) dewatering outlet is closed by closed ball valve, makes container be in air-tight state, in filler, injects appropriate water, open outside temperature regulating device, utilize heating tube to heat, monitored by the first thermometer, make water temperature constant in preset temp A;
6) after outside temperature regulating device opens for some time, by gas temperature and actual temperature B in the second thermometer measuring vessel, by magnetic stirring apparatus control B=37 DEG C, then in filler, inject the humidity in a small amount of water management container, read registration by wet bulb thermometer;
7) after terminating, first turn off external power source control device, outside temperature regulating device and gas-holder pressurizing valve, then adjust pore container internal gas pressure is reduced, open blind flange and take out sample pallet.
As a kind of preferred version, step 3) described when electromagnetic field generator is placed on sample pallet, the single porous cell culture plate being placed with cell sample is placed on sample pallet simultaneously.
The present invention has following beneficial effect relative to prior art:
1, multi-scenarios method In vitro cell experiment device of the present invention can realize the multi-scenarios method of the field of force, magnetic field, electric field by electromagnetic field generator, also the field of force, magnetic field, being used alone of electric field be can realize, more real biomechanics, bioelectromagnetics conveniently can be studied at the Discussion of Mechanism of molecular biology level.
2, multi-scenarios method In vitro cell experiment device of the present invention can control atmosphere, the temperature and humidity condition needed for cell cultures in container, the magnetic stirring apparatus that bottom is installed, being evenly distributed of water temperature can be realized, ensure that the uniform spatial distribution of internal tank experimental temperature, independently can carry out In vitro cell experiment, without the need to putting into specific cell culture incubator.
3, multi-scenarios method In vitro cell experiment device of the present invention is when testing, the wiring of electromagnetic field generator can be connected with external power source control device by line terminal, according to actual needs, utilize external power source control device, the cell culture environment under stationary magnetic field, pulsed magnetic field, steady electric field, constant pressure can be realized, but also be applicable to other and need temperature control to exert oneself research of other experiment such as chemosynthesis of electromagnetic field environment.
4, the sample pallet of multi-scenarios method In vitro cell experiment device use of the present invention, the height of sample table can be regulated, thus obtain suitable space, the cell sample that conveniently can carry out many group different experimental conditions is tested simultaneously, the demand of lot of experiments can be met, and sample table adopts the irregular roundness shape design cropping part arc length, the circuit discharge of convenient electromagnetic generating unit, also can make the range of temperature of thermometer energy measuring vessel different heights, make environmental Kuznets Curves more science.
5, the structure of container of multi-scenarios method In vitro cell experiment device use of the present invention is simple, with low cost, be made up of stainless steel, have the advantages that height is withstand voltage, height is airtight, easily disinfect, and adopt Modular assembled to form, facilitate cleaning and sterilizing and change bad, old component.
Accompanying drawing explanation
Fig. 1 is the container front side structural representation of multi-scenarios method In vitro cell experiment device of the present invention.
Fig. 2 is the container plan structure schematic diagram of multi-scenarios method In vitro cell experiment device of the present invention.
Fig. 3 is the sample pallet Facad structure schematic diagram of multi-scenarios method In vitro cell experiment device of the present invention.
Fig. 4 is sample table surface tissue schematic diagram in the sample pallet of multi-scenarios method In vitro cell experiment device of the present invention.
Fig. 5 is the electromagnetic field generator structural representation with multi-scenarios method In vitro cell experiment matched with devices of the present invention.
Wherein, 1-container, 2-blind flange, 3-magnetic stirring apparatus, 4-sample pallet, 5-filler, 6-dewatering outlet, 7-heating tube, 8-first thermometer hole, 9-ball valve, 10-air inlet port, 11-production well, 12-line terminal, 13-weather gage hole, 14-second thermometer hole, 15-wet bulb thermometer hole, 16-rubber cradle, 17-sample table, 18-thread rack, 19-bracket holes, 20-holes, 21-hex-nut, 22-Helmholtz coil, 23-parallel-plate electrode, the dull and stereotyped pallet of 24-, 25-power supply, 26-terminal stud, 27-porous cell culture plate.
Embodiment
Embodiment 1:
As shown in Figure 1-Figure 3, the multi-scenarios method In vitro cell experiment device of the present embodiment, the blind flange 2, magnetic stirring apparatus 3 and the sample pallet 4 that comprise container 1, match with container 1.
As depicted in figs. 1 and 2, described container 1 is for being equipped with the cylindrical straight tubular of insulating interlayer, it adopts stainless material to make, pressurize can be stablized, the upper end of described container 1 is provided with filler 5, lower end is provided with dewatering outlet 6, heating tube 7 and the first thermometer hole 8, the larger water of specific heat capacity is injected as thermal insulation medium by filler 5, described dewatering outlet 6 place is provided with ball valve 9, first thermometer hole 8 is placed with the first thermometer, and described heating tube 7 is connected with outside temperature regulating device respectively with the first thermometer; The top that described blind flange 2 is connected to container 1 is formed can the structure of opened/closed container 1, its surface is provided with air inlet port 10, production well 11, line terminal 12, thermometer hole 14, weather gage hole 13, second and wet bulb thermometer hole 15, described weather gage hole 13 is placed with weather gage, described second thermometer hole 14 is placed with the second thermometer, described wet bulb thermometer hole 15 is placed with wet bulb thermometer, rubber cradle 16 is provided with, to improve the sealing effectiveness of container 1 between blind flange 2 and container 1 top; Described magnetic stirring apparatus 3 is arranged on the bottom of container 1, and the water of container 1 interlayer can be made to produce eddy current, even to ensure the distribution of water temperature in container.
As shown in Figure 3 and Figure 4, described sample pallet 4 comprises four layers of sample table 17 and three thread racks 18, described every layer of sample table 17 is for cropping the irregular roundness shape of part arc length, its surface is provided with three bracket holes 19 and 16 borings 20, described bracket holes 19 matches with thread rack 18, described boring 20 can place the 15ml centrifuge tube corresponding with its diameter, and described thread rack 18 adopts stainless material to make; By thread rack 18 through bracket holes 19, regulated and fix the height of every layer of sample table 17 by hex-nut 21, described every layer of sample table 17 can place electromagnetic field generator with porous cell culture plate and/or single porous cell culture plate.
As shown in Figure 5, described electromagnetic field generator comprises one group of parallel Helmholtz coil 22, one group of parallel-plate electrode 23, dull and stereotyped pallet 24 and power supply 25, described parallel-plate electrode 23 is placed on dull and stereotyped pallet 24, is provided with terminal stud 26 bottom described power supply 25; When electromagnetic field generator is placed in container 1, the wiring of described terminal stud 26 is connected with external power source control device by line terminal 12.
As Figure 1-Figure 5, the multi-scenarios method In vitro cell experiment process of the present embodiment, comprises the following steps:
1) by after container 1 sterilization and disinfection, in container 1, a certain amount of deionized water is added to keep the humidity environment of cell cultures;
2), between the parallel-plate electrode 23 porous cell culture plate 27 being placed with cell sample being placed on electromagnetic field generator, porous cell culture plate 27 is made to load electromagnetic field;
3) electromagnetic field generator is placed in the first layer (from top to bottom) sample table 17 of sample pallet 4, then sample pallet 4 is placed in container 1, the wiring of terminal stud 26 is connected with external power source control device, by blind flange 2 unopened container 1 by line terminal 12;
4) by air inlet port 10 and band rate of flow meter containing 5%CO
2medical mixed gas gas-holder be connected, then adjust pore 11, monitored by weather gage, obtain stable air pressure;
5) dewatering outlet 6 is closed by closed ball valve 9, makes container 1 be in air-tight state, injects appropriate water, open outside temperature regulating device in filler 5, utilize heating tube 7 to heat, monitored by the first thermometer, make water temperature constant in preset temp A;
6) after outside temperature regulating device opens for some time, by gas temperature and actual temperature B in the second thermometer measuring vessel 1, by magnetic stirring apparatus 3 control B=37 DEG C;
7) after terminating, first turn off external power source control device, outside temperature regulating device and gas-holder pressurizing valve, then adjust pore 11 container 1 internal gas pressure is reduced, open blind flange 2 and take out sample pallet 4.
Embodiment 2:
The multi-scenarios method In vitro cell experiment process principal feature of the present embodiment is: step 3) in, regulate the hex-nut 21 of sample pallet 4, obtain suitable space, electromagnetic field generator is placed in the first layer (from top to bottom) sample table 17 of sample pallet 4, the single porous cell culture plate being placed with cell sample is placed in the second layer sample table 17 of sample pallet 4 simultaneously, namely achieve the cell sample experiment making electricity consumption, magnetic, power multi-scenarios method, also achieve the cell sample experiment being used alone the field of force.All the other are with embodiment 1.
Embodiment 3:
The multi-scenarios method In vitro cell experiment process principal feature of the present embodiment is: step 3) in, regulate the hex-nut 21 of sample pallet 4, obtain suitable space, electromagnetic field generator is placed in the first layer (from top to bottom) sample table 17 of sample pallet 4, the single porous cell culture plate being placed with cell sample is placed in the second layer sample table 17 of sample pallet 4 simultaneously, and the 15ml centrifuge tube being placed with cell sample is placed in the boring 20 of the third layer sample table 17 of sample pallet 4, test while realizing various kinds of cell sample.All the other are with embodiment 1.
The above; be only patent preferred embodiment of the present invention; but the protection domain of patent of the present invention is not limited thereto; anyly be familiar with those skilled in the art in the scope disclosed in patent of the present invention; be equal to according to the technical scheme of patent of the present invention and patent of invention design thereof and replaced or change, all belonged to the protection domain of patent of the present invention.
Claims (7)
1. multi-scenarios method In vitro cell experiment device, is characterized in that: comprise can constant control water temperature container, to match with container and surface be provided with multiple perforate blind flange, the uniform magnetic stirring apparatus of distribution of water temperature can be realized and the sample pallet of the electromagnetic field generator that can place with porous cell culture plate and/or single porous cell culture plate; The top and bottom of described container are respectively arranged with filler and dewatering outlet, and described dewatering outlet place is provided with ball valve, and the top that described blind flange is connected to container is formed can the structure of opened/closed container, and described magnetic stirring apparatus is arranged on the bottom of container; Described sample pallet is placed in container when testing, comprise Multilayer Samples platform and at least three for supporting the thread rack of sample table, described every layer of sample table is the irregular roundness shape cropping part arc length, its surface is provided with at least three bracket holes matched with thread rack, described thread rack is through regulating by hex-nut after bracket holes and the height of fixed sample platform, and the surface of described every layer of sample table is also provided with multiple boring for placing centrifuge tube; One of them perforate on described blind flange is line terminal, described electromagnetic field generator comprises one group of parallel Helmholtz coil, one group of parallel-plate electrode, dull and stereotyped pallet and power supply, described parallel-plate electrode is placed on dull and stereotyped pallet, is provided with terminal stud bottom described power supply; When electromagnetic field generator is placed in container, the wiring of described terminal stud is connected with external power source control device by line terminal.
2. multi-scenarios method In vitro cell experiment device according to claim 1, it is characterized in that: described container lower end is also provided with heating tube and the first thermometer hole for placing the first thermometer, and described heating tube is connected with outside temperature regulating device respectively with the first thermometer.
3. multi-scenarios method In vitro cell experiment device according to claim 2, is characterized in that: the perforate that described blind flange surface is arranged also comprise air inlet port, production well, for place barometric weather gage hole, for place the second thermometer the second thermometer hole and for placing hygroscopic wet bulb thermometer hole.
4. the multi-scenarios method In vitro cell experiment device according to any one of claim 1-3, is characterized in that: described container is the cylindrical straight tubular that insulating interlayer is housed, and it adopts stainless material to make.
5. the multi-scenarios method In vitro cell experiment device according to any one of claim 1-3, is characterized in that: be provided with rubber cradle between described blind flange and the top of container.
6., based on the multi-scenarios method In vitro cell experiment method of device described in claim 3, it is characterized in that comprising the following steps:
1), after being sterilized by container sterilization, in container, a certain amount of deionized water is added to keep the humidity environment of cell cultures;
2) the porous cell culture plate being placed with cell sample is placed between the parallel-plate electrode of electromagnetic field generator;
3) electromagnetic field generator is placed on sample pallet, then sample pallet is placed in container, the wiring of terminal stud is connected with external power source control device by line terminal, by blind flange unopened container;
4) by air inlet port and band rate of flow meter containing 5%CO
2medical mixed gas gas-holder be connected, then adjust pore, monitored by weather gage, obtain stable air pressure;
5) dewatering outlet is closed by closed ball valve, makes container be in air-tight state, in filler, injects appropriate water, open outside temperature regulating device, utilize heating tube to heat, monitored by the first thermometer, make water temperature constant in preset temp A;
6) after outside temperature regulating device opens for some time, by gas temperature and actual temperature B in the second thermometer measuring vessel, by magnetic stirring apparatus control B=37 DEG C;
7) after terminating, first turn off external power source control device, outside temperature regulating device and gas-holder pressurizing valve, then adjust pore container internal gas pressure is reduced, open blind flange and take out sample pallet.
7. multi-scenarios method In vitro cell experiment method according to claim 6, it is characterized in that: step 3) described when electromagnetic field generator is placed on sample pallet, the single porous cell culture plate being placed with cell sample is placed on sample pallet simultaneously.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| FI127032B (en) | 2014-03-21 | 2017-10-13 | Magnasense Tech Oy | Measuring arrangement, apparatus for a measuring arrangement and method for measuring a sample |
| CN105445192A (en) * | 2015-12-14 | 2016-03-30 | 天津港东科技发展股份有限公司 | Embedded magnetic stirring constant-temperature sample rack |
| CN107102217A (en) * | 2017-04-25 | 2017-08-29 | 广东电网有限责任公司电力科学研究院 | A kind of power frequency electromagnetic field animal experiment system |
| CN109596670B (en) * | 2018-12-08 | 2023-07-25 | 北京理工大学 | Comprehensive generation system for electric/magnetic field environment |
| CN111996120A (en) * | 2020-09-25 | 2020-11-27 | 南通纺织丝绸产业技术研究院 | Cell culture device with micro-current stimulation function |
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| CN102653719A (en) * | 2012-04-28 | 2012-09-05 | 吉林大学 | Cell culture device capable of generating multiple magnetic fields and culture method |
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| CN101274101A (en) * | 2008-05-21 | 2008-10-01 | 中国农业大学 | Novel high-density carbon dioxide sterilization reactor |
| CN201271585Y (en) * | 2008-09-26 | 2009-07-15 | 杭州博日科技有限公司 | Constant temperature trough with magnetic stirring |
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