CN209797993U - cell in vitro fluid shear force stimulation culture device - Google Patents
cell in vitro fluid shear force stimulation culture device Download PDFInfo
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- CN209797993U CN209797993U CN201920186844.8U CN201920186844U CN209797993U CN 209797993 U CN209797993 U CN 209797993U CN 201920186844 U CN201920186844 U CN 201920186844U CN 209797993 U CN209797993 U CN 209797993U
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Abstract
In order to realize the circulation culture of cells in a culture solution, realize the simulation of a mode closer to a human body and simultaneously avoid the damage of the culture to the cells, in particular to the damage of the cells in the culture solution, the in vitro fluid shear force stimulation culture device for the cells is realized, the output of the stable gas quantity of a pneumatic pump is used for supplying gas to two culture solution pipes, the circulation of the culture solution is formed by the arrangement of a one-way valve, the switching of the gas supply is realized, and the circulation device which is efficient, has various fluid circulation modes and has no damage to the cells is realized.
Description
Technical Field
The utility model belongs to human cell in vitro culture field discloses a culture solution circulation's the amazing culture apparatus of cell external fluid shear force, provides one kind and has realized the amazing culture apparatus of cell external fluid shear force under clean, not damaged having utilized the atmospheric pressure loading to realize the culture solution circulation.
background
In order to perform related research, various blood flow simulation circulation devices have been put to practical use; meanwhile, in order to simulate various blood flow environments in a human body, the switching method of various flow modes is further applied; however, in practice it has been found that if the cells are placed in suspension in the culture broth, cell death results from the use of an inappropriate pump and from the use of a squeeze against the flow tube. The blood circulation in the human body depends on the pumping of the heart and the function of the heart valve, thereby realizing the circulation of the blood in the human body.
In cell cycle experiments, it is also critical that the pump itself be clean.
SUMMERY OF THE UTILITY MODEL
in order to better realize cell culture under the environment of simulated blood flow in vitro, a cell in vitro fluid shear force stimulation culture device and a method thereof are provided; the device realizes the mode of simulating the blood circulation in vivo as much as possible in vitro, avoids the extrusion to a culture solution circulation pipeline, utilizes the pressure of blood flow to carry out the blood circulation, and simultaneously utilizes the one-way valve to simulate the effect of a heart valve; thereby realizing the fluid shear force stimulation culture of the cells.
The utility model provides a cell in vitro fluid shear force stimulation culture device, which comprises an air supply device, a culture solution flowing device, a parallel flat plate flowing culture cavity and a computer control system; the air supply device comprises a pneumatic pump, a drying bottle, an electric control valve, a filter and a connecting pipeline; the air supply device realizes 2 air supply ports, the 2 air supply ports supply air alternately, and meanwhile, the air supply ports which do not supply air are connected with the atmosphere during alternation; the culture solution flowing device consists of 4 one-way valves, 2 culture solution pipes, a plurality of pipelines and a culture solution pipe fixing support.
Further, the pneumatic pump in the gas supply device is a pump capable of conveying a stable gas amount, and meanwhile, a pump body or a pipeline of a gas flowing part is easy to detach and carry out sterilization treatment, for example: a peristaltic pump, the pump being capable of achieving a flow rate of 0-1000 ml/min; the drying bottle is filled with a drying agent which has no dust and particles and is non-toxic and biologically accessible, such as alumina, or after moisture absorption; the electric control valve realizes switching of gas output by the pneumatic pump to be transmitted to different passages A or passages B in a mode of extruding the channel hose, and can realize connection of one port with the atmosphere when pressure gas is transmitted outwards to the other port.
Further, the culture solution flowing device is composed of the 4 one-way valves, the 2 culture solution pipes and the pipelines, and can realize unidirectional flowing, unidirectional intermittent flowing and reciprocating flowing of the culture solution fluid by matching with the air supply mode of the air supply device; when the air feeder carries out the air feed to a culture solution pipe, the air feed mouth and the atmosphere of another culture solution pipe are connected, set for the time that the air feed mouth switches, air feed mouth switching time be less than the culture solution in the culture solution pipe the time that the culture solution is whole to flow out can.
Furthermore, the culture solution flows in a single direction, and the liquid outlets of the 2 culture solution tubes are respectively connected with a one-way valve in a forward direction and then are connected with the liquid inlet ports of the parallel flat plate flow cavities together; and 2, after the culture solution return ports of the culture solution pipes are respectively connected with a one-way valve in a reverse connection mode, the culture solution return ports are connected with the liquid outlet ports of the parallel flat plate flowing cavities together, the action of compressed gas at the gas supply port on the culture solution in the culture solution pipes is switched, the unidirectional flowing of the culture solution in the parallel flat plate flowing cavities is realized, and unidirectional fluid shear force stimulation is applied to cells cultured in the parallel flat plate flowing cavities.
Further, the unidirectional intermittent flow of the culture solution fluid is realized by controlling the start and stop of the pump, and all connection modes of the culture solution flowing device and the unidirectional flow of the culture solution fluid are the same.
Furthermore, the culture fluid flows in the reciprocating direction, the liquid outlets of 2 culture fluid tubes are respectively connected with two one-way valves in series, the 2 nd one-way valve is connected with the backflow port of the culture fluid tube, the one-way valves are connected in the forward direction, and one culture fluid tube is connected with the liquid inlet port of the parallel flat plate flow cavity after the connecting pipeline between the 2 matched one-way valves is disconnected; the other culture liquid tube is connected with the liquid outlet port of the parallel flat plate flow cavity after the opening of the connecting tube in the middle of the 2 one-way valves; the reciprocating motion of culture solution in the parallel flat plate flowing cavity is realized by matching the action of switching gas supply port compressed gas on the culture solution in the culture solution pipe, and the stimulation of constant reciprocating fluid shear force is applied to cells cultured in the parallel flat plate flowing cavity.
Further, a culture solution pipe fixing bracket in the culture solution flowing device is used for fixing the culture solution pipes to be placed at the same height; the liquid level height of the culture solution in the two culture solution tubes is not required to be the same under the static state, and when the culture solution in one culture solution tube completely flows to the pipeline, the other culture solution tube cannot be fully filled.
Further, the parallel flat plate flow chamber is horizontally arranged; the in vitro cell fluid shear force stimulation culture device is fixed relative to the culture liquid pipe during operation.
furthermore, in order to ensure the gas environment and the temperature environment of the culture, the culture solution flowing device and the parallel flat plate flowing culture cavity can be placed in a constant temperature device; to ensure 5% CO2, the inlet of the pneumatic pump was connected to a gas atmosphere of 5% CO 2.
Furthermore, the utility model discloses still provide the application method of cell in vitro fluid shear force stimulation culture apparatus, including the following step:
1. Sterilizing a gas pipeline, a culture solution flowing device and a parallel flat plate flowing culture cavity in the in vitro cell fluid shear force stimulation culture device;
2. According to the experimental purpose, the culture solution flowing device is connected according to the modes of unidirectional flowing, unidirectional intermittent flowing and reciprocating flowing of culture solution fluid, the pump is used for driving gas to convey the culture solution pipe to push the culture solution, the circulating motion of the culture solution is realized according to different arrangements of the one-way valve, and meanwhile, the effect on cells in the parallel flat plate flowing experimental cavity is realized;
3. Adding the culture solution into a culture solution pipe, opening the electric control valve in the pipe for putting the culture solution, and enabling the air supply device to supply air to enable the culture solution to enter the culture solution flowing device and the parallel flat plate flowing culture cavity;
4. Setting the required fluid shear force in a computer control system according to the fluid shear force required to be loaded;
5. The air inlet of the air supply device is connected, and the air inlet is communicated with carbon dioxide gas with the concentration of 5 percent, and the culture solution flowing device and the parallel flat plate flowing culture cavity are placed in a constant temperature environment;
6. And starting the in vitro fluid shear force stimulation culture device for the cells to carry out experiments.
The beneficial effects of the utility model are as follows
The utility model simplifies the air supply device, and can easily realize high-temperature high-pressure sterilization and disinfection in each link from the air inlet of the pump except using disposable consumables, thereby ensuring that the possibility of cell culture pollution is greatly reduced, and reducing the cleaning cost of the pump;
the utility model realizes a cell culture mode which does not cause damage to cells due to excessive extrusion;
The utility model realizes the function of multiple fluid modes by using the same air pressure supply mode through the simple arrangement mode of changing pipelines and one-way valves;
The utility model realizes that very little culture solution circularly runs to meet the requirement of cell culture, and can realize the collection of cell secretion;
the utility model realizes that the culture solution can circulate indiscriminately, namely the circulation of the circulating solution realizes the full and uniform mixing of the culture in the operation process, and the culture solution of the same part is not only used for stimulating cells during the supply;
the utility model discloses a culture solution that fluidic unidirectional flow, unidirectional intermittent type formula flow, the method that reciprocating direction flows made can be indiscriminately circulate and stimulate and cultivate the cell of parallel dull and stereotyped flow intracavity simultaneously.
Therefore, the utility model provides a cell in vitro fluid shear force stimulation culture device.
Drawings
FIG. 1 is a schematic diagram of a cell in vitro fluid shear force stimulation culture device according to the present invention.
FIG. 2 is a schematic view of a state 1 in which a culture solution flows in one direction and a culture solution flows intermittently in one direction.
FIG. 3 shows the schematic of state 2 of unidirectional flow and unidirectional intermittent flow of culture medium.
FIG. 4 is a schematic view of a state 1 in which a culture solution flows in a reciprocating direction.
FIG. 5 is a schematic view showing the state 2 of the culture solution in the reciprocating direction.
Fig. 6 is a schematic diagram of the state that the electronic control valve squeezes the passage of the hose.
fig. 7 is a schematic diagram of the closed state of the squeezing hose channel of the electric control valve.
fig. 8 is a schematic diagram of two states of the electric control valve.
The reference numbers:
the device comprises a gas supply device (1), a culture solution flowing device (2), a parallel flat plate flowing culture cavity (3) and a computer control system (4);
The drying device comprises a pneumatic pump (1-1), a drying bottle (1-2), an electric control valve (1-3), a filter (1-4) and a connecting pipeline (1-5); an air supply port (A) and an air supply port (B); the device comprises a culture liquid pipe A (2-1), a culture liquid pipe B (2-2), a one-way valve (2-3), a culture liquid pipe fixing support (2-3), an air supply device output air pressure (p 1), an external atmospheric pressure (p0), a closing state (C) of an electric control valve to a hose, a closing state (O) of the electric control valve to the hose, a clamp (1-3-1) and a hose (1-9).
Detailed Description
the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments, and are not intended to limit the scope of the patent application of the present invention; based on the utility model provides an embodiment, all the utility model discloses the equal change or the modification change of accomplishing under method and the technical scheme that indicate all should belong to the utility model discloses the patent range that covers.
It is noted that when an element is referred to as being "secured to" another element, it can be directly on the other element or intervening elements may also be present; when a component is referred to as being "connected" to another component, it can be directly connected to the other component or intervening components may also be present; the terms "upper", "lower", "left", "right" and the like are used herein for illustrative purposes only.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs; the terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention; as used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
A cell in vitro fluid shear force stimulation culture device comprises an air supply device (1), a culture solution flowing device (2), a parallel flat plate flowing culture cavity (3) and a computer control system (4); the air supply device (1) comprises a pneumatic pump (1-1), a drying bottle (1-2), an electric control valve (1-3), a filter (1-4) and a connecting pipeline (1-5); the air supply device (1) realizes 2 air supply ports, wherein the 2 air supply ports comprise an air supply port (A) and an air supply port (B) for alternately supplying air, and meanwhile, the air supply port which does not supply air is connected with the atmosphere during alternation; the culture solution flowing device (2) consists of 4 one-way valves (2-2), 2 culture solution pipes (2-1), a plurality of pipelines and culture solution pipe fixing supports (2-3).
Further, the pneumatic pump is a pump capable of delivering a stable amount of gas, and a pump body or a pipeline of a gas flowing portion is easily detached and sterilized, for example: a peristaltic pump; the pump can supply air with the flow rate of 0-3000ml/min, and the air supply of the pump is adjusted according to the requirement of culture solution circulation; the drying bottle (1-2) is filled with a drying agent which has no dust and particles and is non-toxic and biologically accessible, such as alumina, or after moisture absorption; the electric control valve realizes switching of gas output by the pneumatic pump to be transmitted to different passages A or passages B in a mode of extruding the channel hose, and can realize connection of one port with the atmosphere when pressure gas is transmitted outwards to the other port.
Furthermore, the culture solution flowing device (2) is composed of the 4 one-way valves (2-3), the 2 culture solution pipes (the culture solution pipe A (2-1) and the culture solution pipe B (2-2)) and a plurality of pipelines, and can realize unidirectional flowing, unidirectional intermittent flowing and reciprocating flowing of the culture solution fluid by matching with the gas supply mode of the gas supply device (1); when the air feeder supplies air to the culture solution tube A, the air supply port of the culture solution tube B is connected to the atmosphere, and the time for switching the air supply port is set, wherein the time for switching the air supply port is shorter than the time for the whole content in the culture solution tube to flow out.
Furthermore, the culture fluid flows in a single direction, the liquid outlet of each of 2 culture fluid tubes (culture fluid tube A (2-1) and culture fluid tube B (2-2)) is connected with a one-way valve (2-3) and then is connected with the liquid inlet of the parallel flat plate flow chamber (3) in a forward and backward direction, the liquid outlet of each of 2 culture fluid tubes (culture fluid tube A (2-1) and culture fluid tube B (2-2)) is connected with the liquid return of the one-way valve (2-3) in a reverse direction and then is connected with the liquid outlet of the parallel flat plate flow chamber (3) in a reverse direction, the effect of compressed gas at the gas supply port on the culture fluid in the culture fluid tubes is switched, the culture fluid flows in the parallel flat plate flow chamber (3) in a single direction, and the cells cultured in the parallel flat plate flow chamber are stimulated by the fluid shear force in the single direction.
Further, the unidirectional intermittent flow of the culture solution fluid is realized by controlling the start and stop of the pump (1-1), and all connection modes of the culture solution flowing device (2) and the unidirectional flow of the culture solution fluid are the same.
Furthermore, the culture fluid flows in the reciprocating direction, the liquid outlets of 2 culture fluid tubes are respectively connected with two one-way valves in series, the 2 nd one-way valve is connected with the backflow port of the culture fluid tube, the one-way valves are connected in the forward direction, and one culture fluid tube is connected with the liquid inlet port of the parallel flat plate flow cavity after the connecting pipeline between the 2 matched one-way valves is disconnected; the other culture liquid tube is connected with the liquid outlet port of the parallel flat plate flow cavity after the opening of the connecting tube in the middle of the 2 one-way valves; the reciprocating motion of culture solution in the parallel flat plate flowing cavity is realized by matching the action of switching gas supply port compressed gas on the culture solution in the culture solution pipe, and the stimulation of constant reciprocating fluid shear force is applied to cells cultured in the parallel flat plate flowing cavity.
Further, a culture solution pipe fixing bracket (2-4) in the culture solution flowing device (2) is used for fixing the culture solution pipe to be placed at the same height; the liquid level height of the culture solution in the two culture solution tubes is not required to be the same under the static state, and when the culture solution in one culture solution tube completely flows to the pipeline, the other culture solution tube cannot be fully filled.
further, the parallel flat plate flow cavity (3) is horizontally arranged; the in vitro cell fluid shear force stimulation culture device is fixed relative to the culture liquid pipe during operation.
Further, in order to ensure the gas environment and the temperature environment of the culture, the culture solution flowing device and the parallel flat plate flowing culture cavity are placed in a constant temperature device; to ensure 5% CO2, the inlet of the pneumatic pump was connected to a gas atmosphere of 5% CO 2.
Further, according to the utility model discloses still provide the application method of cell in vitro fluid shear force stimulation culture apparatus, including the following step:
1. Sterilizing a gas pipeline, a culture solution flowing device and a parallel flat plate flowing culture cavity in the in vitro cell fluid shear force stimulation culture device;
2. According to the experimental purpose, the culture solution flowing device is connected according to the modes of unidirectional flowing, unidirectional intermittent flowing and reciprocating flowing of culture solution fluid, the pump is used for driving gas to convey the culture solution pipe to push the culture solution, the circulating motion of the culture solution is realized according to different arrangements of the one-way valve, and meanwhile, the effect on cells in the parallel flat plate flowing experimental cavity is realized;
3. adding the culture solution into a culture solution pipe, opening the electric control valve in the pipe for putting the culture solution, and enabling the air supply device to supply air to enable the culture solution to enter the culture solution flowing device and the parallel flat plate flowing culture cavity;
4. Setting the required fluid shear force in a computer control system according to the fluid shear force required to be loaded;
5. The air inlet of the air supply device is connected, and the air inlet is communicated with carbon dioxide gas with the concentration of 5 percent, and the culture solution flowing device and the parallel flat plate flowing culture cavity are placed in a constant temperature environment;
6. and starting the in vitro fluid shear force stimulation culture device for the cells to carry out experiments.
The above description is for the detailed description of the preferred possible embodiments of the present invention, but the embodiments are not intended to limit the scope of the present invention, and all equivalent changes or modifications accomplished by the methods suggested by the present invention should fall within the scope of the present invention.
Claims (6)
1. a cell in vitro fluid shear force stimulation culture device is characterized by comprising an air supply device, a culture solution flowing device, a parallel flat plate flowing culture cavity and a computer control system; the air supply device comprises a pneumatic pump, a drying bottle, an electric control valve, a filter and a connecting pipeline; the air supply device realizes 2 air supply ports, the 2 air supply ports supply air alternately, and meanwhile, the air supply ports which do not supply air are connected with the atmosphere during alternation; the culture solution flowing device consists of 4 one-way valves, 2 culture solution pipes, a plurality of pipelines and a culture solution pipe fixing support.
2. The culture apparatus for stimulating the shear force in vitro cells according to claim 1, wherein the pneumatic pump of the gas supply apparatus is a pump capable of delivering a stable amount of gas, and the pump or the pipeline of the gas flowing portion is easily disassembled and sterilized, for example: a peristaltic pump, the pump being capable of achieving a flow rate of 0-1000 ml/min; the drying bottle is filled with a drying agent which has no dust and particles and is non-toxic and biologically accessible, such as alumina, or after moisture absorption; the electric control valve realizes switching of gas output by the pneumatic pump to be transmitted to different passages A or passages B in a mode of extruding the channel hose, and can realize connection of one port with the atmosphere when pressure gas is transmitted outwards to the other port.
3. the culture apparatus for stimulating cell in vitro by fluid shear force according to claim 1, wherein the culture fluid flowing apparatus comprises the 4 one-way valves, 2 culture fluid pipes and a plurality of pipelines, and can realize unidirectional flow, unidirectional intermittent flow and reciprocating flow of the culture fluid by matching with the air supply manner of the air supply apparatus; when the air supply device supplies air to one culture solution pipe, the air supply port of the other culture solution pipe is connected with the atmosphere, and the switching time of the air supply port is set, wherein the switching time of the air supply port is shorter than the time of the culture solution in the culture solution pipe flowing out completely; the culture solution flows in a single direction, and the liquid outlets of the 2 culture solution pipes are respectively connected with a one-way valve in a forward direction and then are connected with the liquid inlet ports of the parallel flat plate flow cavities together; the culture solution return ports of the 2 culture solution pipes are respectively connected with a one-way valve for reverse connection, and then are connected with the liquid outlet ports of the parallel flat plate flowing cavities together, the action of compressed gas at the gas supply port on the culture solution in the culture solution pipes is switched, the unidirectional flowing of the culture solution in the parallel flat plate flowing cavities is realized, and unidirectional fluid shear force stimulation is applied to cells cultured in the parallel flat plate flowing cavities; the culture solution fluid intermittently flows in a single direction, the intermittent flow is realized by controlling the starting and stopping of the pump, and all connection modes of the culture solution flowing device and the culture solution fluid flowing in the single direction are the same; the culture solution flows in the reciprocating direction, the liquid outlets of 2 culture solution pipes are respectively connected with two one-way valves in series, the 2 nd one-way valve is connected with the backflow port of the culture solution pipe, the one-way valves are connected in the forward direction, and one culture solution pipe is connected with the liquid inlet port of the parallel flat plate flow cavity together after the connecting pipeline between the 2 matched one-way valves is disconnected; the other culture liquid tube is connected with the liquid outlet port of the parallel flat plate flow cavity after the opening of the connecting tube in the middle of the 2 one-way valves; the reciprocating motion of culture solution in the parallel flat plate flowing cavity is realized by matching the action of switching gas supply port compressed gas on the culture solution in the culture solution pipe, and the stimulation of constant reciprocating fluid shear force is applied to cells cultured in the parallel flat plate flowing cavity.
4. The device for culturing cells in vitro under the stimulation of fluid shear force according to claim 1, wherein the culture solution tube fixing bracket of the culture solution flowing device is used for fixing the culture solution tubes to be placed at the same height; the liquid level height of the culture solution in the two culture solution tubes is not required to be the same under the static state, and when the culture solution in one culture solution tube completely flows to the pipeline, the other culture solution tube cannot be fully filled.
5. The device according to claim 1, wherein the parallel plate flow chamber is horizontally disposed and fixed relative to the culture solution tube during operation.
6. The in vitro fluid shear force stimulation culture device for the cells according to claim 1, wherein the culture gas environment and the temperature environment are cultured, and the culture solution flowing device and the parallel plate flowing culture chamber are arranged in a constant temperature device; the inlet of the pneumatic pump was connected to a gas atmosphere of 5% CO 2.
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| CN201920186844.8U CN209797993U (en) | 2019-02-02 | 2019-02-02 | cell in vitro fluid shear force stimulation culture device |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109735448A (en) * | 2019-02-02 | 2019-05-10 | 上海天引生物科技有限公司 | A kind of cells in vitro hydrodynamic shear stimulation culture apparatus and its method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109735448A (en) * | 2019-02-02 | 2019-05-10 | 上海天引生物科技有限公司 | A kind of cells in vitro hydrodynamic shear stimulation culture apparatus and its method |
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