CN210030739U - Cell stretching culture device with adjustable basement membrane rigidity - Google Patents
Cell stretching culture device with adjustable basement membrane rigidity Download PDFInfo
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- CN210030739U CN210030739U CN201920833355.7U CN201920833355U CN210030739U CN 210030739 U CN210030739 U CN 210030739U CN 201920833355 U CN201920833355 U CN 201920833355U CN 210030739 U CN210030739 U CN 210030739U
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Abstract
A cell stretching and culturing device with adjustable basement membrane rigidity mainly solves the problems that a cell culturing device in the prior art is high in use cost and large in operation difficulty. It is including cultivateing the board, cultivateing lid and base, cultivate board detachably and fix in the base top, form an airtight space between cultivateing board and the base, cultivate the lid and place in the cultivation board top, be equipped with the several on the cultivation board and cultivate the hole, cultivate the hole bottom and be equipped with compound elasticity basement membrane, the outside fixedly connected with aspiration pump of base and the pump of admitting air, aspiration pump and the pump of admitting air communicate airtight space respectively, aspiration pump and the pump of admitting air electricity respectively are connected with the controller. Its advantage does: simple structure is small and exquisite, and preparation low cost, and the user can select the culture plate that has different rigidity basilar membranes as required, and culture plate and base be detachable construction, are convenient for wash and cyclic utilization, realize the tensile/compression function to the cell through the cooperation of aspiration pump, air inlet pump and automatically controlled pneumatic valve.
Description
Technical Field
The utility model relates to a cell breeding device, concretely relates to cell stretch breeding device with adjustable basement membrane rigidity.
Background
Cell tension is the force of the interaction of the basement membrane or extracellular matrix with adherent cells. The mechanical interaction between the cell and the extracellular matrix plays an important role in regulating and controlling the proliferation, differentiation, migration, apoptosis and the like of the cell. In the field of cell research, the application method of tension force is mainly as follows: stretching of silica gel film and atomic force microscope technology. Both of these techniques have significant disadvantages: firstly, they are very costly and difficult to handle, and secondly, when they are used to apply force to cells, they cannot be used simultaneously with other stimuli (such as drugs, hypoxic stimuli, etc.), resulting in a significant limitation of their range of use. The method is characterized in that a silica gel film stretching method only can stretch cells without compression or other stress stimulation forms, an atomic force microscope only can perform pressure stimulation on single cells, the change of protein and nucleic acid in the cells after the pressure stimulation needs at least 5^105 cell amount, which is difficult to complete by the atomic force microscope, in addition, the several ways of applying force stimulation are that the cells are cultured in a common cell culture plate, the hardness of the culture plate per se also has mechanical stimulation on the cells, the stress stimulation on the cells by the culture plates with different hardness is different, and the influence of the factors is not considered by the several mechanical stimulation devices. In addition, the human body has many cells subjected to stress stimulation in a tissue site where nutrition is deficient, such as articular cartilage cells, nucleus pulposus cells of intervertebral disc, which are in a state of being anoxic for a long time, and the cells cannot be tested by a common culture plate.
SUMMERY OF THE UTILITY MODEL
In order to overcome the not enough of background art, the utility model provides a cell culture apparatus that leads with adjustable basement membrane rigidity solves the problem that cell culture apparatus use cost among the prior art is high, the operation degree of difficulty is big.
The utility model discloses the technical scheme who adopts: the utility model provides a cell culture apparatus that leads with adjustable basement membrane rigidity, includes culture plate, cultivates lid and base, culture plate detachably fixes in the base top, forms an airtight space between culture plate and the base, cultivate the lid and place in culture plate top, be equipped with the several on the culture plate and cultivate the hole, cultivate the hole bottom and be equipped with compound elasticity basement membrane, the outside fixedly connected with aspiration pump of base and the pump of admitting air, aspiration pump and the pump of admitting air communicate airtight space respectively, aspiration pump and the pump of admitting air are connected with the controller respectively to the electricity.
The composite elastic basement membrane is formed by sequentially compounding a PA gel layer, a PVC elastic membrane and a rubber sealing membrane.
The air suction pump and the air inlet pump are communicated with the closed space through hoses, and the hoses are provided with electric control air valves which are electrically connected with the controller.
The cultivation device further comprises a detachable sealing box, and the cultivation plate, the cultivation cover and the base are placed in the detachable sealing box.
The detachable sealing box comprises a box body and a box cover, wherein the box cover is covered above the box body, a first sealing ring is arranged at the joint of the box cover and the box body, the sealing ring is fixedly connected in the box cover, a U-shaped notch is formed in one side of the box body, and a second sealing ring is arranged in the U-shaped notch.
The culture plate is buckled and connected on the base.
The utility model has the advantages that: simple structure is small and exquisite, and preparation low cost, and the user can select the culture plate that has different rigidity basilar membranes as required, and culture plate and base be detachable construction, are convenient for wash and cyclic utilization, realize the tensile/compression function to the cell through the cooperation of aspiration pump, air inlet pump and automatically controlled pneumatic valve.
Drawings
FIG. 1 is a schematic structural diagram of a cell stretching and cultivating device with adjustable basement membrane rigidity according to an embodiment of the present invention;
fig. 2 is a schematic structural diagram of a cell stretching and culturing device with adjustable basement membrane rigidity according to an embodiment of the invention;
FIG. 3 is a schematic structural diagram of the cell stretching and cultivating device with adjustable basement membrane rigidity according to the embodiment of the present invention after air intake;
fig. 4 is a schematic structural diagram of the cell stretching cultivation device with adjustable basement membrane rigidity after air exhaust;
fig. 5 is a schematic structural diagram of a composite elastic basement membrane of a cell stretching culture device with adjustable basement membrane rigidity according to an embodiment of the invention;
fig. 6 is a schematic structural diagram of a second sealing ring of a cell stretching and culturing device with adjustable basement membrane rigidity according to an embodiment of the present invention;
fig. 7 is a schematic diagram illustrating the operation of a cell stretching and cultivating device with adjustable basement membrane rigidity according to an embodiment of the present invention.
In the figure: 1-culture plate, 2-culture cover, 3-base, 4-culture hole, 5-composite elastic basement membrane, 6-air pump, 7-air inlet pump, 8-controller, 51-protein coating, 52-PA gel layer, 53-PVC elastic membrane, 54-rubber sealing membrane, 81-electric control air valve, 91-box body, 92-box cover, 93-first sealing ring and 94-second sealing ring.
Detailed Description
The embodiments of the present invention will be further explained with reference to the accompanying drawings:
as shown in fig. 1-7, a cell stretching cultivation device with adjustable basement membrane rigidity comprises a culture plate 1, a culture cover 2 and a base 3, wherein the culture plate 1 is detachably fixed above the base 3, a closed space is formed between the culture plate 1 and the base 3, the culture cover 2 is placed above the culture plate 1, a plurality of culture holes are arranged on the culture plate 1, a composite elastic basement membrane 5 is arranged at the bottom of each culture hole, an air suction pump 6 and an air inlet pump 7 are fixedly connected to the outside of the base 3, the air suction pump 6 and the air inlet pump 7 are respectively communicated with the closed space, the air suction pump 6 and the air inlet pump 7 are respectively and electrically connected with a controller 8, a user controls the air inlet pump 7/the air suction pump 6 to work through the controller 8, selects the culture plate 1 with proper rigidity according to cell requirements during use, cleans and sterilizes the culture plate 1, and then fixes the culture plate 1 above the base 3, coating a protein coating 51 on the composite elastic basement membrane 5 in the culture hole, implanting cells into the culture hole, covering the culture cover 2 after the culture is finished, utilizing negative pressure generated by air suction of a closed space and positive pressure generated by air inflation to deform the composite elastic basement membrane 5, and adjusting the pressure of air suction/air exhaust through the controller 8 to change the deformation amount of the basement membrane so as to enable the cells growing adherent to be stimulated by traction/compression traction stress; the controller 8 is in the prior art, and comprises a control chip, a display screen, a control button, a power module and a timing module, wherein the control chip, the display screen, the control button, the power module and the timing module are all connected with a circuit board in the controller 8, the circuit board is provided with a corresponding control circuit, a user inputs data such as the working cycle times, the single air intake/air exhaust duration and the working interval time of the air intake pump 7/air exhaust pump 6 through the control button and displays the data by a display, the control chip controls the air intake pump 7/air exhaust pump 6 to work according to the data input by the control button and the time data fed back by the timing module, and the power module supplies power for electronic elements of the cultivating; the model of the air intake pump 7 and/or the air suction pump 6 is MZP-AM-265, and the air suction pump and the air intake pump can be directly placed on one side of the base, so that the air suction pump and the air intake pump are convenient to disassemble; the number of culture wells on the culture plate is 6.
When the rigidity of the surrounding environment of the cell is different, the response of the cell to the same magnitude of mechanics is different, so that it is necessary to design basement membranes with different rigidities.
Preferably, the composite elastic basement membrane 5 is formed by sequentially compounding a PA gel layer 52, a PVC elastic membrane 53 and a rubber sealing membrane 54; composite elastic base film 5 the procedure for preparation of PA gel solution was prepared by varying the concentration of acrylamide (3% -5%) and bisacrylamide (0.01% -0.15%) in deionized water, initiating polymerization with 0.05% ammonium persulfate and 0.1% N, N' -Tetramethylethylenediamine (TEMED), pipetting 15 μ l of the polymerized solution onto 0.5% 3-aminopropyltrimethoxysilane and 1% glutaraldehyde treated glass coverslips to form PA gel layer 52; photoactivation of the PA gel layer 52 by sulfo-SANPAH under UV light (10 min), followed by incubation of the PA gel layer 52 in 200 μ g/mL laminin solution at 37 ℃ for 4 h, followed by rinsing in Phosphate Buffered Saline (PBS) and UV sterilization for 30 min; the Young modulus (100 Pa, 500Pa, 1KPa and 5 KPa) of PA gel is quantified by AFM, the concentration ratio is improved, so that PA gel layers 52 with different rigidities are formed, then the PA gel layers 52, the PVC elastic membrane 53 and the rubber sealing membrane 54 are sequentially adhered by glue, the adhered composite elastic basement membrane 5 is adhered to the bottom of the culture plate 1 by glue, culture plates 1 with different rigidities are formed, and a user can select the culture plate 1 with the proper basement membrane rigidity to culture cells according to needs.
Preferably, the air pump 6 and the air intake pump 7 are communicated with the closed space through a hose, an electric control air valve 81 is arranged on the hose, the electric control air valve 81 is electrically connected with the controller 8, the electric control air valve 81 is normally closed, when the closed space is subjected to air intake/air exhaust, the controller 8 controls the electric control air valve 81 to be powered on to enable the closed space to be communicated with the air intake pump 7/the air pump 6, and after the air intake/air exhaust is finished, the controller 8 controls the electric control air valve 81 to be powered off to enable the closed space to be isolated from the air intake pump 7/the air pump 6, so that the air leakage of the closed space is avoided.
Preferably, the culture device further comprises a detachable sealing box, the culture plate 1, the culture cover 2 and the base 3 are placed in the detachable sealing box, and because a gap is formed between the culture cover 2 and the culture plate 1 and air is circulated, bacteria in the culture hole can grow normally, when certain cells, such as cartilage cells in joints, nucleus pulposus cells of intervertebral discs and the like, need to be cultured, the cells are firstly implanted into the culture hole, then the culture plate 1 and the base 3 are installed, the culture cover 2 is covered, then the whole culture plate is placed in the detachable sealing box, and the cells are placed for several hours to slowly consume oxygen in the detachable sealing box, so that an oxygen-poor stimulation function is realized; the detachable sealing box, the culture plate and the culture cover are all made of transparent materials such as PVC, PMMA and the like.
Preferably, the detachable sealing box comprises a box body 91 and a box cover 92, the box cover 92 covers the box body 91, a first sealing ring 93 is arranged at the joint of the box cover 92 and the box body 91, the sealing ring is fixedly connected in the box cover 92, a U-shaped notch is formed in one side of the box body 91, a second sealing ring 94 is arranged in the U-shaped notch, the U-shaped notch is used for placing a lead of the controller 8, the air intake pump 7, the air suction pump 6 and the electric control air valve 81, the controller 8 can be placed outside the sealing box, the controller 8 can be operated under the oxygen-deficient environment without influencing cells, and the second sealing ring 94 is used for sealing the U-shaped notch, so that the influence of the lead of the controller 8 on the sealing performance of the detachable sealing box is reduced.
Preferably, the culture plate 1 is connected to the base 3 in a snap-fit manner, so that the culture plate 1 can be conveniently detached by the snap-fit connection manner, the situation that the composite elastic base film 5 is damaged due to friction during assembly can be avoided, and the service life of the composite elastic base film 5 is prolonged.
When the culture plate is used, firstly, the culture plate 1 required by an experiment is selected for cleaning and disinfection, then the culture plate 1 is buckled on the base 3, the composite elastic basement membrane 5 in the culture hole is coated with the protein coating 51, cells are implanted in the culture hole, the culture cover 2 is covered after the completion, then the whole culture plate is placed in the detachable sealing box, the controller 8 is placed outside the detachable sealing box, the lead of the controller 8 penetrates through the second sealing ring 94 to be electrically connected with the air inlet pump 7 and the electric control air valve 81 of the air suction pump 6, then the single working time, the interval time, the duration time and the cycle time of the air inlet pump 7 and the air suction pump 6 are input from the control button in the controller 8, the specific control process is as follows, the control chip controls the air inlet pump 7 to intake N seconds and controls the electric control air valve 81 on the air inlet hose to be electrified, and after the air inlet is finished, the control chip simultaneously closes the air inlet pump 7 and the, and stopping N minutes, then controlling the air suction pump 6 to suck air for N seconds by the control chip, controlling the electric control air valve 81 on the air suction hose to be electrified, closing the air suction pump 6 and the electric control air valve 81 on the air suction hose at the same time by the control chip after the air suction is finished, and stopping N minutes, so as to circulate until the circulation frequency reaches the preset frequency.
The beneficial effects of this embodiment: simple structure is small and exquisite, and preparation low cost, and the user can select the culture plate that has different rigidity basilar membranes as required, and culture plate and base be detachable construction, are convenient for wash and cyclic utilization, realize the tensile/compression function to the cell through the cooperation of aspiration pump, air inlet pump and automatically controlled pneumatic valve.
The examples should not be construed as limiting the invention, but any modifications made based on the spirit of the invention are intended to be within the scope of the invention.
Claims (6)
1. A basement membrane rigidity adjustable cell stretch breeding device which characterized in that: including cultivateing board (1), cultivateing lid (2) and base (3), cultivate board (1) detachably and fix in base (3) top, form an airtight space between cultivateing board (1) and base (3), cultivate lid (2) and place in cultivateing board (1) top, be equipped with the several on cultivateing board (1) and cultivate hole (4), cultivate hole (4) bottom and be equipped with compound elasticity basement membrane (5), base (3) outside fixedly connected with aspiration pump (6) and air intake pump (7), aspiration pump (6) and air intake pump (7) communicate airtight space respectively, aspiration pump (6) and air intake pump (7) are connected with controller (8) respectively.
2. The basement membrane stiffness-adjustable cell stretching culture device of claim 1, wherein: the composite elastic basement membrane (5) is formed by sequentially compounding a PA gel layer (52), a PVC elastic membrane (53) and a rubber sealing membrane (54).
3. The basement membrane stiffness-adjustable cell stretching culture device of claim 1, wherein: the air suction pump (6) and the air inlet pump (7) are communicated with the closed space through hoses, an electric control air valve (81) is arranged on each hose, and the electric control air valve (81) is electrically connected with the controller (8).
4. The basement membrane stiffness-adjustable cell stretching culture device of claim 1, wherein: the cultivation device further comprises a detachable sealing box, and the cultivation plate (1), the cultivation cover (2) and the base (3) are placed in the detachable sealing box.
5. The basement membrane stiffness-adjustable cell stretching culture device of claim 4, wherein: can dismantle seal box includes box body (91) and lid (92), lid (92) lid closes in box body (91) top, and lid (92) are equipped with first sealing washer (93) with the junction of box body (91), sealing washer fixed connection is in lid (92), and box body (91) one side is equipped with the U-shaped breach, is equipped with second sealing washer (94) in the U-shaped breach.
6. The basement membrane stiffness-adjustable cell stretching culture device of claim 1, wherein: the culture plate (1) is connected to the base (3) in a buckling mode.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201920833355.7U CN210030739U (en) | 2019-06-04 | 2019-06-04 | Cell stretching culture device with adjustable basement membrane rigidity |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201920833355.7U CN210030739U (en) | 2019-06-04 | 2019-06-04 | Cell stretching culture device with adjustable basement membrane rigidity |
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| Publication Number | Publication Date |
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| CN210030739U true CN210030739U (en) | 2020-02-07 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201920833355.7U Expired - Fee Related CN210030739U (en) | 2019-06-04 | 2019-06-04 | Cell stretching culture device with adjustable basement membrane rigidity |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111849771A (en) * | 2020-07-31 | 2020-10-30 | 中国人民解放军空军军医大学 | Culture device for carrying out negative pressure and mechanical stimulation on cell-containing culture |
| WO2023042670A1 (en) * | 2021-09-16 | 2023-03-23 | 国立大学法人北海道大学 | Cell culture container, cell culture kit, and cell culture method |
-
2019
- 2019-06-04 CN CN201920833355.7U patent/CN210030739U/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111849771A (en) * | 2020-07-31 | 2020-10-30 | 中国人民解放军空军军医大学 | Culture device for carrying out negative pressure and mechanical stimulation on cell-containing culture |
| CN111849771B (en) * | 2020-07-31 | 2023-03-28 | 中国人民解放军空军军医大学 | Culture device for carrying out negative pressure and mechanical stimulation on cell-containing culture |
| WO2023042670A1 (en) * | 2021-09-16 | 2023-03-23 | 国立大学法人北海道大学 | Cell culture container, cell culture kit, and cell culture method |
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| Date | Code | Title | Description |
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| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20200207 |