CN102127506B - High-flux culture device for cell traction stimulation - Google Patents
High-flux culture device for cell traction stimulation Download PDFInfo
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- CN102127506B CN102127506B CN 201010583804 CN201010583804A CN102127506B CN 102127506 B CN102127506 B CN 102127506B CN 201010583804 CN201010583804 CN 201010583804 CN 201010583804 A CN201010583804 A CN 201010583804A CN 102127506 B CN102127506 B CN 102127506B
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Abstract
The invention discloses a high-flux culture device for cell traction stimulation, comprising a first organic glass plate and a second organic glass plate. Through holes are arranged on the first organic glass plate and the second organic glass plate, a pellosil is arranged between the first organic glass plate and the second organic glass plate, a third organic glass plate is arranged at the lower part of the second organic glass plate, a first organic glass frame is arranged between the second organic glass plate and the third organic glass plate, an organic glass hollow cavity is formed by the second organic glass plate, the third organic glass plate and the first organic glass frame; both sides of the organic glass hollow cavity are respectively connected with pressure control units through channels; and fine holes are arranged at both sides of the first organic glass frame to communicate the interior of the organic glass hollow cavity and the channels. The device can finely adjust the traction amount of a film, realize periodic traction and realize microscopic observation in a traction process after the device is inverted and a transparent tray is arranged.
Description
Technical field
The present invention relates to biomedical engineering field, particularly relate to a kind of high-flux culture device for the cell traction stimulation.
Background technology
Modern biological study shows, the outside physiological environment can cellular function exert an influence, and wherein mechanics factor plays vital effect to growth, differentiation, development and the immune response of cell.Therefore study cell and be subjected to how to change mechanical stimulation into intracellular electrochemical signals behind the tractive, especially Skeletal Muscle Cell and myocardial cell are received publicity by the reaction behind the tractive increasingly.
Cell is carried out the principle of tractive, generally be to use the bio-compatibility such as silica gel good, less, the resilient material that is easy to deformation, thereon culturing cell of self Young's modulus simultaneously, after cell is close to growth, silica gel material is stretched, thereby drive cell traction.At present existing various structures is used for the research that cell traction stimulates, and can be summarized as two class schemes: a class is to utilize the nested structures such as stainless steel coil manual operations to realize that the pellosil Uniform Tension comes the tractive cell; Another kind ofly then mainly rely on the cell culture apparatus that computer control machinery device makes silica gel material and do whole tractive.Yet this two classes scheme cuts both ways: first kind scheme needs manual regulation cell traction degree, can accomplish meticulous adjusting amount of tension, but is only applicable to static tractive experiment, and is difficult to property performance period tractive.The Equations of The Second Kind scheme has commercially produced product, can property performance period tractive, and it is high to repeat precision and the consistence of tractive, but its cost is extremely expensive, be limited by again structure self, can not under the object lens inverted microscope that cell cultures is commonly used, observe and carry out high throughput analysis.
Summary of the invention
The technical problem that (one) will solve
The first technical problem that the present invention will solve is how to realize periodicity tractive and the high throughput analysis of cell under the prerequisite of saving cost;
The second technical problem that the present invention will solve is how the periodicity traction process of cell to be carried out microscopic examination easily.
(2) technical scheme
For solving the problems of the technologies described above, a kind of high-flux culture device for the cell traction stimulation is provided, comprise: the first poly (methyl methacrylate) plate and vertical pile are in the second poly (methyl methacrylate) plate of described the first poly (methyl methacrylate) plate bottom, has the corresponding through hole in position on described the first poly (methyl methacrylate) plate and described the second poly (methyl methacrylate) plate, between the upper surface of the lower surface of described the first poly (methyl methacrylate) plate and described the second poly (methyl methacrylate) plate, be provided with pellosil, the bottom of described the second poly (methyl methacrylate) plate is provided with the 3rd poly (methyl methacrylate) plate, be provided with the first synthetic glass frame between described the second poly (methyl methacrylate) plate and described the 3rd poly (methyl methacrylate) plate, described the second poly (methyl methacrylate) plate, described the 3rd poly (methyl methacrylate) plate and described the first synthetic glass frame form the synthetic glass cavity, the both sides of described synthetic glass cavity are connected with pressure control unit by the passage that plexi-glass tubular forms respectively, and the both sides of the first synthetic glass frame are provided with pore and are communicated with described synthetic glass cavity inside and passage.
Preferably, also comprise transparent pallet, and described high-flux culture device is set to be inverted in formation inversion type high-flux culture device on the transparent pallet.
Preferably, described transparent pallet comprises the 4th poly (methyl methacrylate) plate and is pasted on the second synthetic glass frame on described the 4th poly (methyl methacrylate) plate.
Preferably, the cross-sectional dimension of described the first poly (methyl methacrylate) plate, the second poly (methyl methacrylate) plate and described the 3rd poly (methyl methacrylate) plate is 127mm * 85mm, the thickness of described the first poly (methyl methacrylate) plate is 6mm, the thickness of described the second poly (methyl methacrylate) plate, described the 3rd poly (methyl methacrylate) plate and described the first synthetic glass frame is 2mm, and the sidewall width of described the first synthetic glass frame is 2.5mm.
Preferably, described through hole is circular hole, and radius is 6mm, is 5 * 4 arranged.
Preferably, the thickness of described pellosil is 0.254mm, and its cross-sectional dimension is greater than the cross-sectional dimension of described the first poly (methyl methacrylate) plate.
Preferably, the diameter of described pore is 1mm, and the external diameter of described passage is 6mm, and internal diameter is 4mm.
Preferably, described pressure control unit comprises: signal generator, in the two paths of signals that sends with described signal generator, the first via connects the first electromagnetic relay, the first liquid storage tank and compressed gas cylinder successively, and the second the tunnel connects the second electromagnetic relay and the second liquid storage tank successively by phase inverter.
Preferably, between described the first electromagnetic relay and described the first liquid storage tank, be provided with the first valve, between described the second electromagnetic relay and the second liquid storage tank, be provided with the second valve.
Preferably, described the first valve is connected with the passage of synthetic glass cavity one side, described the second valve is connected with the passage of synthetic glass cavity opposite side, connecting path at described the first valve and described passage is provided with the first flow meter, is provided with the second under meter at the connecting path of described the second valve and described the second liquid storage tank.
(3) beneficial effect
High-flux culture device of the present invention uses the double-deck synthetic glass AND DEWATERING FOR ORIFICE STRUCTURE that is embedded with pellosil, the upper aperture plate array culturing cell of pellosil, and the lower cavity transmission pressure drives.Apply simultaneously periodic pressure by using single pressure source that the top all cells is cultivated the unit, thereby cell traction is realized unified control.This device also can be realized being inverted contact tractive pattern by being equipped with transparent pallet, makes things convenient for device to carry out microscopic examination in the cell traction process.Compare with the easy device of nested steel ring etc., not only can meticulous adjusting film stretching amount, can also property performance period tractive, and remarkable increase does not occur in cost.Compare with computer control machinery device, can realize high throughput analysis, after will installing inversion and being equipped with transparent pallet, can also realize the microscopic examination in the traction process.This device is made relatively simple, and complete function has important practical value and industrialization prospect simultaneously.
Description of drawings
Fig. 1 (a) is the structural representation according to the high-flux culture device that is used for the cell traction stimulation of one embodiment of the invention;
Fig. 1 (b) is the structural representation according to the high-flux culture device that is used for the cell traction stimulation of another embodiment of the present invention;
Fig. 2 (a)-Fig. 2 (c) is respectively the structural representation according to the first (the second) poly (methyl methacrylate) plate of the high-flux culture device that is used for the cell traction stimulation of the embodiment of the invention, synthetic glass frame, the 3rd poly (methyl methacrylate) plate;
Fig. 3 is the structural representation according to the pressure control unit of the high-flux culture device that is used for the cell traction stimulation of another embodiment of the present invention;
Fig. 4 (a) and Fig. 4 (b) are the application schematic diagram according to the high-flux culture device that is used for the cell traction stimulation of another embodiment of the present invention.
Wherein, the 1, first poly (methyl methacrylate) plate; 2, the second poly (methyl methacrylate) plate; 3, pellosil; 4, the 3rd poly (methyl methacrylate) plate; 5, passage; 6, the first synthetic glass frame; 7, pore; 8, high-flux culture device; 9, transparent pallet; 10, be inverted high-flux culture device; 11, signal generator; 12, the first electromagnetic relay, 13, phase inverter; 14, the second electromagnetic relay; 15, the signalling system of hydrostatic control; 16, compressed gas cylinder; 17, pressurized gas direction; 18, the first liquid storage tank; 19, the first valve; 20, first flow meter; 21, the direction of liquid access to plant; 22, the direction of device expel liquid; 23, the second valve; 24, the second under meter; 25, the second liquid storage tank; 26, gas is discharged direction.
Embodiment
Below in conjunction with drawings and Examples, the specific embodiment of the present invention is described in further detail.Following examples are used for explanation the present invention, but are not used for limiting the scope of the invention.
Shown in Fig. 1 (a), one embodiment of the invention provides a kind of high-flux culture device for the cell traction stimulation, comprise: the first poly (methyl methacrylate) plate 1, the second poly (methyl methacrylate) plate 2 and pellosil 3, the first poly (methyl methacrylate) plate 1 and the second poly (methyl methacrylate) plate 2 vertical pile, has the corresponding through hole in position on the first poly (methyl methacrylate) plate 1 and the second poly (methyl methacrylate) plate 2, pellosil 3 is arranged between the upper surface of the lower surface of the first poly (methyl methacrylate) plate 1 and the second poly (methyl methacrylate) plate 2, the bottom of the second poly (methyl methacrylate) plate 2 is provided with the 3rd poly (methyl methacrylate) plate 4, be provided with the first synthetic glass frame 6 between the second poly (methyl methacrylate) plate 2 and described the 3rd poly (methyl methacrylate) plate 4, described the second poly (methyl methacrylate) plate 2, described the 3rd poly (methyl methacrylate) plate 4 and described the first synthetic glass frame 6 form the synthetic glass cavity, the both sides of described synthetic glass cavity are connected with pressure control unit by the passage 5 that plexi-glass tubular forms respectively, and the both sides of the first synthetic glass frame 6 are provided with pore 7 and are communicated with described synthetic glass cavity inside and passage 5.
High-flux culture device among this embodiment can realize as follows, at first, and sandwich structure of the second poly (methyl methacrylate) plate 2 bonding formation of the first poly (methyl methacrylate) plate 1, one deck pellosil 3 and the layer that one deck is thicker; Then, get out pore 7 on the wall with the first synthetic glass frame 6 of a rectangle and form the sidewall of synthetic glass cavitys, and paste on the 3rd poly (methyl methacrylate) plate 4 as this synthetic glass cavity lower wall; Afterwards the bottom of the second poly (methyl methacrylate) plate 2 and sidewall that is the first synthetic glass frame 6 of cavity are bonded together, at last passage 5 is bonded on the sidewall and pore 7 UNICOMs, form the passage that the synthetic glass cavity is connected to pressure control unit, thereby form a kind of high-flux culture device 8, cell will be incubated at pellosil 3 towards the one side of the first poly (methyl methacrylate) plate 1.
Shown in Fig. 1 (b), another embodiment of the present invention provides a kind of high-flux culture device for the cell traction stimulation, comprise: the first poly (methyl methacrylate) plate 1, the second poly (methyl methacrylate) plate 2 and pellosil 3, the first poly (methyl methacrylate) plate 1 and the second poly (methyl methacrylate) plate 2 vertical pile, has the corresponding through hole in position on the first poly (methyl methacrylate) plate 1 and the second poly (methyl methacrylate) plate 2, pellosil 3 is arranged between the upper surface of the lower surface of the first poly (methyl methacrylate) plate 1 and the second poly (methyl methacrylate) plate 2, the bottom of the second poly (methyl methacrylate) plate 2 is provided with the 3rd poly (methyl methacrylate) plate 4, be provided with the first synthetic glass frame 6 between the second poly (methyl methacrylate) plate 2 and described the 3rd poly (methyl methacrylate) plate 4, described the second poly (methyl methacrylate) plate 2, described the 3rd poly (methyl methacrylate) plate 4 and described the first synthetic glass frame 6 form the synthetic glass cavity, the both sides of described synthetic glass cavity are connected with pressure control unit by the passage 5 that plexi-glass tubular forms respectively, and the both sides of the first synthetic glass frame 6 are provided with pore 7 and are communicated with described synthetic glass cavity inside and passage 5.In addition, this high-flux culture device also comprises the transparent pallet 9 that places under the first poly (methyl methacrylate) plate 1.Transparent pallet 9 comprises the 4th poly (methyl methacrylate) plate and is pasted on the second synthetic glass frame on the 4th poly (methyl methacrylate) plate.
High-flux culture device among this embodiment can be realized as follows: make high-flux culture device 8 according to the implementation method among the first embodiment first, other gets two the second synthetic glass frames of size, nested bonding to forms transparent pallet 9 on the 4th poly (methyl methacrylate) plate, and upside down high-flux culture device 8 thereon, then finish a kind of making of the inversion high-flux culture device 10 that stimulates for cell traction.Wherein, two the second synthetic glass frames are used for preventing that nutrient solution from overflowing rear pickup outside atmosphere from internal layer synthetic glass frame.It is in order to make pellosil heave rear bottom near transparent pallet 9, to be positioned at the following microscope of bottom of device to the micro-Real Time Observation of cell traction process thereby be beneficial to that high-flux culture device 8 is inverted.
Shown in Fig. 2 (a) and Fig. 2 (c), the cross-sectional dimension of the first poly (methyl methacrylate) plate 1, the second poly (methyl methacrylate) plate 2 and the 3rd poly (methyl methacrylate) plate 4 all is preferably 127mm * 85mm, the thickness of the first poly (methyl methacrylate) plate 1 is preferably 6mm, and the thickness of the second poly (methyl methacrylate) plate 2 and the 3rd poly (methyl methacrylate) plate 4 all is preferably 2mm.Through hole on the first poly (methyl methacrylate) plate 1 and the second poly (methyl methacrylate) plate 2 is circular hole, and radius is preferably 6mm, is 5 * 4 arranged.The thickness of pellosil 3 is 0.254mm, and its cross-sectional dimension is greater than the cross-sectional dimension of the first poly (methyl methacrylate) plate 1.The first synthetic glass frame 6 of synthetic glass cavity externally measured is of a size of long 127mm, wide 85mm, highly be 2mm, sidewall width is 2.5mm, and the diameter of the pore 7 that gets out on the first synthetic glass frame 6 is 1mm, be connected to the external diameter 6mm of the passage 5 of external pressure control unit on the sidewall, internal diameter 4mm is shown in Fig. 2 (b).
Be illustrated in figure 3 as the structural representation of the hydraulic control unit of the embodiment of the invention, the signal that signal generator 11 is sent is divided into two-way, one road signal is directly controlled the first electromagnetic relay 12, another road is then through control the second electromagnetic relay 14 behind the phase inverter 13, keep the first electromagnetic relay 12 and the second electromagnetic relay 14, the first valve 19 and the second valve 23 to be in all the time on off state reverse, signal generator 11, the first electromagnetic relay 12, phase inverter 13 and the second electromagnetic relay 14 form the signalling system 15 of hydrostatic control thus.The mode of operation of hydraulic control system is: signalling system 15 is sent signal, and the second valve 23 is closed so that the first valve 19 is opened, gas enters the first liquid storage tank 18 by compression gas direction 17 from compressed gas cylinder 16, thereby its internal pressure is increased, then liquid extrudes from the first liquid storage tank 18, through the first valve 19 and first flow meter 20, the cavity that enters high-flux culture device 8 or be inverted high-flux culture device 10 along the direction 21 of liquid access to plant applies pressure to it; After the signal upset of signalling system 15, the first valve 19 is closed and the second valve 23 is opened, high-flux culture device 8 or be inverted pressure in high-flux culture device 10 cavitys greater than direction 22 after everywhere, therefore liquid enters the second liquid storage tank 25 along the direction 22 of device expel liquid through the second valve 23 and the second under meter 24, and discharge direction 26 to extraneous Exhaust Gas by gas, thereby discharge pressure.In this way repeatedly, hydraulic control system can accomplish to apply periodically back and forth pressure to being inverted high-flux culture device 10.The signal frequency of conditioning signal producer 11 can be controlled the cycle that applies reciprocal pressure, regulates the gas flow rate of compressed gas cylinder 16 releases and can control the peak value size that applies reciprocal pressure.
The function that is used for the cell traction stimulation of the embodiment of the invention is used as shown in Figure 4, to be inverted high-flux culture device 10 as example, when liquid enters the cavity of being inverted high-flux culture device 10 and applies when strong along the direction 21 of liquid access to plant, the Young's modulus of pellosil 3 is less, obvious expansion deformation occurs, thereby make the cell of cultivating on the first poly (methyl methacrylate) plate 1 pellosil 3 surfaces simultaneously be subject to tractive, shown in Fig. 4 (a); When liquid is discharged from be inverted high-flux culture device 10 along the direction 22 of expel liquid, be inverted the cavity pressure of high-flux culture device 10 and reply normal pressure, then the deformation of pellosil 3 is eliminated, thereby remove the cell traction of cultivating on pellosil 3 surfaces of the first poly (methyl methacrylate) plate 1 one side, shown in Fig. 4 (b).
As can be seen from the above embodiments; the high-flux culture device that is used for the cell traction stimulation of the embodiment of the invention uses the double-deck synthetic glass orifice plate that embeds pellosil; pellosil top orifice plate culturing cell; and the below orifice plate plays support and provide protection to pellosil; the pellosil gravitate little when vertical provide protection particularly evident, guarantee to be incubated at the interference that can not influence each other of cell between each vestibule.The orifice plate array of this cell traction high-flux culture device and pellosil are common to use a cavity, so uses single pressure source just can carry out tractive to all cell unifications of cultivating the unit, thereby realizes high throughput analysis.By being equipped with extra pressure control unit, pressure can be passed in real time the top and cultivate the unit, and easilier come mechanically the control pressure exporting change, thus property performance period tractive.Append a transparent pallet or culture dish and just can realize being inverted contact tractive pattern, satisfy the focusing range of Bioexperiment object lens inverted microscope commonly used, thus can be to realize the micro-Real Time Observation in the cell traction process.Should be used for the preparation structure of the high-flux culture device that cell traction stimulates and simple for process, and can greatly reduce difficulty of processing and the cost of device, be applied to biomedical conveniently, and can finally satisfy the industrialization demand.
The above only is preferred implementation of the present invention; should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the technology of the present invention principle; can also make some improvement and modification, these improve and modification also should be considered as protection scope of the present invention.
Claims (9)
1. one kind is used for the high-flux culture device that cell traction stimulates, it is characterized in that, comprise: the first poly (methyl methacrylate) plate (1) and vertical pile are in second poly (methyl methacrylate) plate (2) of described the first poly (methyl methacrylate) plate (1) bottom, has the corresponding through hole in position on described the first poly (methyl methacrylate) plate (1) and described the second poly (methyl methacrylate) plate (2), between described the first poly (methyl methacrylate) plate (1) and described the second poly (methyl methacrylate) plate (2), be provided with pellosil (3), the bottom of described the second poly (methyl methacrylate) plate (2) is provided with the 3rd poly (methyl methacrylate) plate (4), be provided with the first synthetic glass frame (6) between described the second poly (methyl methacrylate) plate (2) and described the 3rd poly (methyl methacrylate) plate (4), described the second poly (methyl methacrylate) plate (2), described the 3rd poly (methyl methacrylate) plate (4) and described the first synthetic glass frame (6) form the synthetic glass cavity, the both sides of described synthetic glass cavity are connected with pressure control unit by the passage (5) that plexi-glass tubular forms respectively, the both sides of described the first synthetic glass frame (6) are provided with pore (7) and are communicated with described synthetic glass cavity inside and passage (5), described pressure control unit comprises signal generator (11), in the two paths of signals that described signal generator (11) sends, first via signal connects the first electromagnetic relay (12) successively, the first liquid storage tank (18) and compressed gas cylinder (16), the second road signal connects the second electromagnetic relay (14) and the second liquid storage tank (25) successively by phase inverter (13).
2. be used for as claimed in claim 1 the high-flux culture device that cell traction stimulates, it is characterized in that, also comprise transparent pallet (9), and described high-flux culture device is set to be inverted in the upper inversion type high-flux culture device that forms of transparent pallet (9).
3. be used for as claimed in claim 2 the high-flux culture device that cell traction stimulates, it is characterized in that, described transparent pallet (9) comprises the 4th poly (methyl methacrylate) plate and is pasted on the second synthetic glass frame on described the 4th poly (methyl methacrylate) plate.
4. be used for as claimed in claim 3 the high-flux culture device that cell traction stimulates, it is characterized in that, the cross-sectional dimension of described the first poly (methyl methacrylate) plate (1), the second poly (methyl methacrylate) plate (2) and described the 3rd poly (methyl methacrylate) plate (4) is 127mm * 85mm, the thickness of described the first poly (methyl methacrylate) plate (1) is 6mm, and the thickness of described the second poly (methyl methacrylate) plate (2), described the 3rd poly (methyl methacrylate) plate (4) and described the first synthetic glass frame (6) is 2mm, and the sidewall width of described the first synthetic glass frame (6) is 2.5mm.
5. be used for as claimed in claim 1 the high-flux culture device that cell traction stimulates, it is characterized in that, described through hole is circular hole, and radius is 6mm, is 5 * 4 arranged.
6. be used for as claimed in claim 1 the high-flux culture device that cell traction stimulates, it is characterized in that, the thickness of described pellosil (3) is 0.254mm, and its cross-sectional dimension is greater than the cross-sectional dimension of described the first poly (methyl methacrylate) plate (1).
7. be used for as claimed in claim 1 the high-flux culture device that cell traction stimulates, it is characterized in that, the diameter of described pore (7) is 1mm, and the external diameter of described passage (5) is 6mm, and internal diameter is 4mm.
8. the high-flux culture device that stimulates for cell traction as described in each such as claim 1-7, it is characterized in that, between described the first electromagnetic relay (12) and described the first liquid storage tank (18), be provided with the first valve (19), between described the second electromagnetic relay (14) and the second liquid storage tank (25), be provided with the second valve (23).
9. be used for as claimed in claim 8 the high-flux culture device that cell traction stimulates, it is characterized in that, described the first valve (19) is connected with the passage of synthetic glass cavity one side, described the second valve (23) is connected with the passage of synthetic glass cavity opposite side, connecting path at the described passage (5) of described the first valve (19) and synthetic glass cavity one side is provided with first flow meter (21), is provided with the second under meter (24) at the connecting path of described the second valve (23) and described the second liquid storage tank (25).
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| CN 201010583804 CN102127506B (en) | 2010-12-07 | 2010-12-07 | High-flux culture device for cell traction stimulation |
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| CN 201010583804 CN102127506B (en) | 2010-12-07 | 2010-12-07 | High-flux culture device for cell traction stimulation |
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| CN102127506A CN102127506A (en) | 2011-07-20 |
| CN102127506B true CN102127506B (en) | 2013-04-17 |
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| JP2001524830A (en) * | 1997-05-10 | 2001-12-04 | バデル，アウグスティヌス | Apparatus for culturing or treating cells |
| CN2818484Y (en) * | 2005-07-15 | 2006-09-20 | 中国人民解放军第四军医大学 | Cell tractive tension controller |
| CN101842474A (en) * | 2007-08-30 | 2010-09-22 | 哈佛学院董事会 | Compliant surface multi-well culture plate |
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2010
- 2010-12-07 CN CN 201010583804 patent/CN102127506B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001524830A (en) * | 1997-05-10 | 2001-12-04 | バデル，アウグスティヌス | Apparatus for culturing or treating cells |
| CN2818484Y (en) * | 2005-07-15 | 2006-09-20 | 中国人民解放军第四军医大学 | Cell tractive tension controller |
| CN101842474A (en) * | 2007-08-30 | 2010-09-22 | 哈佛学院董事会 | Compliant surface multi-well culture plate |
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| JP特表2001524830A 2001.12.04 |
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| 席雨涛 等.培养心肌细胞牵张刺激装置的建立及应用.《第四军医大学学报》.2003,第24卷(第3期),285-286. |
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