Simulated cell culture device in low-oxygen environment

Abstract

The utility model discloses a hypoxia environment simulation cell culture device which comprises a culture box, a supporting plate is arranged in the center of the interior of the culture box, heating plates are arranged on the left inner wall and the right inner wall of the culture box respectively, a placing ring is arranged in the middle of the upper end of the supporting plate, a culture dish is arranged in the placing ring, and the hypoxia environment simulation cell culture device further comprises a stirring mechanism and a humidifying mechanism. The stirring mechanism comprises a stirrer, a magnetic rotor and a rotating shaft, the rotating shaft is rotatably connected to the middle of the bottom wall of the culture dish, the magnetic rotor is fixedly connected to the upper end of the rotating shaft, and the stirrer is arranged in the culture dish; according to the low-oxygen environment simulation cell culture device, when the low-oxygen environment is simulated to culture cells, non-contact stirring of a culture solution is achieved, it is ensured that a cell suspension or a culture medium is evenly mixed, and the environment requirement for cell culture is better met.

Description

TECHNICAL FIELD

[0001] The utility model relates to low oxygen environment simulation cell culture technical field, concretely is low oxygen environment simulation cell culture device. BACKGROUND

[0002] Low oxygen environment usually refers to oxygen concentration is lower than normal physiological level, such as <5%O2, has important significance in cell biology research, can simulate in-vivo tissue microenvironment such as tumor core, stem cell niche etc., is used for studying cell metabolism, proliferation, apoptosis, angiogenesis, signal path regulation process etc., low oxygen environment simulation is the important tool of cell biology research, needs to select suitable method such as gas method vs. chemical method according to experimental purpose, and strictly controls variable, along with the development of technology, dynamic low oxygen culture system such as periodic oxygen concentration fluctuation simulation in-vivo circulation is becoming new trend, can more accurately simulate physiological or pathological microenvironment, low oxygen environment simulation cell culture device is a kind of equipment for simulating in-vivo low oxygen condition in laboratory, is widely used in tumor research, stem cell culture, ischemic disease model construction etc.

[0003] The existing authorized publication number CN103160434A belongs to cell culture technical field, and specifically discloses a simulation in-vivo environment cell automatic culture device, which comprises a laminar flow sterile sealed chamber, wherein the sealed chamber is provided with an internal environment simulation tissue organ culture device and a target cell induction culture device.The device can simulate the low-oxygen and low-osmotic environment for culturing embryonic stem cells, the internal environment during the development and maturation of reproductive cells, the thymus environment for inducing the development and maturation of T cells, the cerebrospinal fluid environment for inducing the differentiation of neural stem cells, and the internal environment of the tissue organ environment such as liver, pancreas, bone marrow and tumor for inducing the differentiation, development and maturation of corresponding stem cells, so as to induce and culture specific high-quality cells to meet the needs of cell basic research and application, and the simulated in-vivo environment also lays a foundation for studying the growth, differentiation and maturation mechanism of the cells in the body, is beneficial to improving the cell culture quality, significantly increasing the repeatability of cytological research, saving the cell culture cost, accelerating the clinical transformation and application of cell culture technology, and providing a good experimental platform for basic researchers.

[0004] The existing low oxygen environment simulation cell culture device has the following problems: when simulating low oxygen environment to culture cells, the in-vivo physiological environment cannot be better simulated, and the uniform mixing of cell suspension or culture medium cannot be ensured, therefore, the low oxygen environment simulation cell culture device is proposed. UTILITY MODEL CONTENT

[0005] The utility model wants to solve the technical problem of overcoming the defects of the prior art, provide low oxygen environment simulation cell culture device, when cultivating the cell in the simulation low oxygen environment, realize the non contact stirring of culture solution, ensure that cell suspension or culture medium is uniformly mixed, better satisfy the environmental demand of cell culture, can effectively solve the problem in the background art.

[0006] In order to realize the above-mentioned purpose, the utility model provides the following technical scheme: low oxygen environment simulation cell culture device, including incubator, the inside center of incubator is equipped with support board, and the left and right inner walls of incubator are equipped with heating plate respectively, and the upper end middle part of support board is equipped with placing ring, and the inside of placing ring is equipped with petri dish, still including stirring mechanism and humidification mechanism;

[0007] Stirring mechanism: it includes stirrer, magnetic rotor and rotating shaft, the middle part of incubator bottom wall is rotatably connected with rotating shaft, and the upper end of rotating shaft is fixedly connected with magnetic rotor, and the inside of petri dish is equipped with stirrer,

[0008] Humidification mechanism: it includes ultrasonic humidifier, airway, rotating column and blade, and the left and right sides of incubator bottom wall are equipped with ultrasonic humidifier respectively, and the upper end of ultrasonic humidifier is equipped with airway respectively, and the left and right inner walls of airway are rotatably connected with evenly distributed blades through rotating column respectively, when cultivating the cell in the simulation low oxygen environment, realize the non contact stirring of culture solution, ensure that cell suspension or culture medium is uniformly mixed, better satisfy the environmental demand of cell culture.

[0009] Further, the right side of the front end of the incubator is provided with a single-chip microcomputer, the input end of the single-chip microcomputer is electrically connected with an external power supply, the input ends of the heating plate and the ultrasonic humidifier are electrically connected with the output end of the single-chip microcomputer, and each electric appliance is electrically connected.

[0010] Further, the stirring mechanism further includes a worm gear, a worm and a motor one, the middle part of the incubator bottom wall is provided with a protective shell, the outer wall of the rotating shaft is fixedly sleeved with the worm gear, the right side between the front and rear inner walls of the protective shell is rotatably connected with the worm, the worm gear is meshed with the worm, the front end of the incubator is provided with the motor one, the output shaft rear end of the motor one is fixedly connected with the front end of the worm, the input end of the motor one is electrically connected with the output end of the single-chip microcomputer, and the rotation is connected.

[0011] Further, the humidification mechanism further includes a chain, a chain wheel and a motor two, the left side outer wall of the rotating column is fixedly connected with two chain wheels respectively, the longitudinally adjacent two chain wheels located in the same vertical plane are drivingly connected through the chain respectively, the front side of the right end of the airway is provided with the motor two respectively, the output shaft left end of the motor two is fixedly connected with the right end of the rotating column of the foremost end transversely adjacent, the input end of the motor two is electrically connected with the output end of the single-chip microcomputer, and the rotation is connected.

[0012] Further, the left and right inner walls and the rear wall of the incubator are respectively provided with temperature sensors and humidity sensors, and the upper end of the incubator is provided with a pressure sensor, and the temperature sensor, the pressure sensor and the humidity sensor are all bidirectionally electrically connected with the single-chip microcomputer, so as to provide detection in the incubator.

[0013] Further, the upper end of the incubator is provided with an air inlet, and the middle part of the air inlet is respectively provided with an air inlet pipe, and the middle part of the air inlet pipe is respectively connected with a mass flow controller, and the mass flow controller is bidirectionally electrically connected with the single-chip microcomputer, so as to facilitate air intake.

[0014] Further, the middle part of the front end of the incubator is hingedly connected with a cover plate, and the sampling hole in the middle part of the front end of the cover plate is inserted with a rubber sealing plug column, so as to facilitate sampling.

[0015] Compared with the prior art, the low-oxygen environment simulation cell culture device has the following advantages:

[0016] 1. The inside of the incubator is pre-heated and temperature-controlled by the heating plate, the ultrasonic humidifier generates tiny mist particles that float upward through the airway, and the rotation column, chain wheel and chain transmission driven by the motor II drives the blade to rotate, adjusts the mist diffusion speed, realizes uniform humidity distribution, and better simulates the physiological environment in the body

[0017] 2. The rotation shaft and the magnetic rotor are driven to rotate synchronously by the worm and the meshed worm wheel under the drive of the motor I, the rotating magnetic field of the magnetic rotor penetrates the branch plate, forms magnetic coupling with the stirrer in the culture dish, drives the stirrer to rotate, realizes contactless stirring of the culture solution, and ensures uniform mixing of the cell suspension or culture medium BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 It is a structural schematic view of the utility model;

[0019] Figure 2 It is a sectional view structural schematic view of the utility model;

[0020] Figure 3 It is a front side sectional view structural schematic view of the utility model;

[0021] Figure 4 It is an upper side sectional view structural schematic view of the utility model;

[0022] Figure 5 It is an enlarged structural schematic view of the utility model A.

[0023] In the figure: 1 incubator, 2 support plate, 3 culture dish, 4 placing ring, 5 stirring mechanism, 51 stirring rod, 52 magnetic rotor, 53 rotating shaft, 54 worm gear, 55 worm, 56 motor one, 6 protective shell, 7 humidifying mechanism, 71 ultrasonic humidifier, 72 airway, 73 rotating column, 74 blade, 75 chain, 76 sprocket, 77 motor two, 8 heating plate, 9 temperature sensor, 10 air inlet pipe, 11 mass flow controller, 12 pressure sensor, 13 single-chip microcomputer, 14 rubber sealing plug column, 15 cover plate, 16 humidity sensor. DETAILED DESCRIPTION

[0024] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0025] Please refer to Figures 1-5The embodiment provides a technical scheme: a low-oxygen environment simulation cell culture device, which comprises a culture box 1, a support plate 2 arranged in the center of the inside of the culture box 1, heating plates 8 arranged on the left and right inner walls of the culture box 1, a placing ring 4 arranged at the upper end of the support plate 2, a culture dish 3 arranged in the inside of the placing ring 4, a stirring mechanism 5 and a humidifying mechanism 7, a single-chip microcomputer 13 arranged at the right side of the front end of the culture box 1, an input end of the single-chip microcomputer 13 electrically connected with an external power supply, output ends of the single-chip microcomputer 13 electrically connected with input ends of the heating plates 8 and an ultrasonic humidifier 71, temperature sensors 9 and humidity sensors 16 arranged on the left and right inner walls and the back wall of the culture box 1, a pressure sensor 12 arranged at the upper end of the culture box 1, the temperature sensors 9, the pressure sensor 12 and the humidity sensors 16 bidirectionally electrically connected with the single-chip microcomputer 13, air inlet pipes 10 arranged at the air inlets of the upper end of the culture box 1, mass flow controllers 11 arranged in series at the middle portions of the air inlet pipes 10, the mass flow controllers 11 bidirectionally electrically connected with the single-chip microcomputer 13, a cover plate 15 hingedly connected with the middle portion of the front end of the culture box 1, sealing rubber strips arranged at the inner walls of the taking and placing holes of the culture box 1 and the rear side edge of the cover plate 15, respectively, for sealing the leakage of the placed gas, L-shaped clamping strips arranged at the left and right sides of the front end of the culture box 1, clamping strips symmetrically arranged on the lower side of the front end of the cover plate 15 and rotatably connected with the rotating shafts, the clamping strips being clamped with the transversely adjacent L-shaped clamping strips, respectively, a rubber sealing plug column 14 inserted into the sampling hole of the middle portion of the front end of the cover plate 15, when simulating a low-oxygen environment for cell culture, the cover plate 15 is opened, the culture dish 3 containing cells is placed into the inside of the placing ring 4, the stirring rod 51 is placed into the inside of the culture dish 3, then the cover plate 15 is closed, then different gases are injected into the inside of the culture box 1 through the air inlet pipes 10, the different gases pass through the mass flow controllers 11, the single-chip microcomputer 13 preprogrammedly controls the mass flow controllers 11 to accurately inject the corresponding gas, the pressure sensor 12 monitors the air pressure in the culture box 1 in real time, when the air pressure changes due to the injection of the gas, the single-chip microcomputer 13 adjusts the air inlet amount or triggers the exhaust valve to maintain constant pressure, the gas is discharged from the exhaust port of the back wall of the culture box 1, the temperature sensor 9 feeds back the temperature data in the culture box 1 to the single-chip microcomputer 13 in real time, when it is necessary to sample the cells in the culture dish 3, the rubber sealing plug column 14 is taken down, the hand is put into the sampling hole to sample the cells, and the rubber sealing plug column 14 is quickly inserted into the inside of the sampling hole after sampling to reduce the gas leakage.

[0026] The stirring mechanism 5 comprises a stirring rod 51, a magnetic rotor 52 and a rotating shaft 53, the middle part of the bottom wall of the incubator 1 is rotationally connected with the rotating shaft 53, the upper end of the rotating shaft 53 is fixedly connected with the magnetic rotor 52, the inside of the culture dish 3 is provided with the stirring rod 51, the stirring mechanism 5 further comprises a worm gear 54, a worm 55 and a motor one 56, the middle part of the bottom wall of the incubator 1 is provided with the protective shell 6, the outer wall of the rotating shaft 53 is fixedly sleeved with the worm gear 54, the right side between the front and rear inner walls of the protective shell 6 is rotationally connected with the worm 55, the worm gear 54 is meshingly connected with the worm 55, the front end of the incubator 1 is provided with the motor one 56, the rear end of the output shaft of the motor one 56 is fixedly connected with the front end of the worm 55, the input end of the motor one 56 is electrically connected with the output end of the single-chip microcomputer 13, then the motor one 56 operates, the output shaft of the motor one 56 drives the worm 55 to rotate, the worm 55 drives the rotating shaft 53 and the magnetic rotor 52 to synchronously rotate through the meshing worm gear 54, the rotating magnetic field of the magnetic rotor 52 penetrates the branch plate 2 and forms magnetic coupling with the stirring rod 51 in the culture dish 3, drives the stirring rod 51 to rotate, realizes non-contact stirring of the culture solution and ensures that the cell suspension or the culture medium is uniformly mixed.

[0027] The humidifying mechanism 7 comprises ultrasonic humidifiers 71, air passages 72, rotating columns 73 and blades 74, and the left and right sides of the bottom wall of the incubator 1 are respectively provided with ultrasonic humidifiers 71, the upper ends of the ultrasonic humidifiers 71 are respectively provided with air passages 72, the left and right inner walls of the air passages 72 are respectively connected through the rotating columns 73, and the blades 74 are uniformly distributed, the humidifying mechanism 7 further comprises chains 75, chain wheels 76 and motors two 77, the left outer wall of the rotating column 73 is respectively fixedly connected with two chain wheels 76, the longitudinally adjacent and located in the same vertical plane two chain wheels 76 are respectively connected through the chain 75, the front side of the right end of the air passage 72 is respectively provided with the motor two 77, the left end of the output shaft of the motor two 77 is respectively fixedly connected with the right end of the foremost rotating column 73 transversely adjacent, the input end of the motor two 77 is electrically connected with the output end of the single-chip microcomputer 13, when the temperature is lower than the set value, the single-chip microcomputer 13 controls the operation of the heating plate 8, the heating plate 8 preheats the inside of the incubator 1, and the heating plate 8 is automatically turned off when the temperature reaches the set value, when it is needed to humidify the inside of the incubator 1, first, the water pipe passing through the outer wall of the incubator 1 injects pure water into the inside of the ultrasonic humidifier 71, then the ultrasonic humidifier 71 operates by adjusting the single-chip microcomputer 13, the ultrasonic humidifier 71 converts electrical energy into mechanical energy by using the transducer at the bottom of the water tank, emits 1.7MHz ultrasonic waves into the water through electronic super frequency oscillation, the high-frequency ultrasonic waves form strong micro-vibration on the liquid surface, and water molecules are broken into fine mist particles with a diameter of only 1-5 microns, these tiny mist particles can be suspended in the air for a long time, the tiny mist particles float upward through the air passage 72, the motor two 77 operates by adjusting the single-chip microcomputer 13, the output shaft of the motor two 77 drives the foremost rotating column 73 to rotate, the foremost rotating column 73 drives the blades 74 to rotate through the transmission of the chain wheel 76 and the chain 75, the mist diffusion speed is adjusted, the humidity is uniformly distributed, and the physiological environment in vivo is better simulated.

[0028] The working principle of the low-oxygen environment simulation cell culture device is as follows: when the cell culture in the simulated low-oxygen environment, first, the cover plate 15 is opened, the culture dish 3 containing cells is placed in the inside of the placing ring 4, the stirrer 51 is placed in the inside of the culture dish 3, then the cover plate 15 is closed, then different gases are injected into the inside of the incubator 1 through the gas inlet pipe 10, the different gases will pass through each mass flow controller 11, the mass flow controller 11 is accurately injected with the corresponding gas through the single-chip microcomputer 13 preset program control, the pressure sensor 12 monitors the gas pressure in the incubator 1 in real time, when the gas injection causes the pressure change, the single-chip microcomputer 13 adjusts the gas inlet amount or triggers the exhaust valve to maintain constant pressure, the gas is discharged from the exhaust port of the rear wall of the incubator 1, the temperature sensor 9 feeds back the temperature data in the incubator 1 to the single-chip microcomputer 13 in real time, when the temperature is lower than the set value, the single-chip microcomputer 13 controls the operation of the heating plate 8, the heating plate 8 preheats the inside of the incubator 1, and the heating plate 8 is automatically closed after the temperature reaches the set value, when it is needed to humidify the inside of the incubator 1, first, the water pipe passing through the outer wall of the incubator 1 injects pure water into the inside of the ultrasonic humidifier 71, the humidity sensor 16 detects the humidity in the incubator 1, the incubator 1 transmits the detection data to the single-chip microcomputer 13, then the ultrasonic humidifier 71 operates through the regulation and control of the single-chip microcomputer 13, the ultrasonic humidifier 71 converts electric energy into mechanical energy by using the transducer at the bottom of the water tank, emits 1.7MHz ultrasonic waves into water through electronic super frequency oscillation, the high-frequency ultrasonic waves form strong micro-vibration on the liquid surface, so that the water molecules are broken into fine mist particles with a diameter of only 1-5 microns, these tiny mist particles can be suspended in the air for a long time, the tiny mist particles float upward through the airway 72, the motor two 77 operates through the regulation and control of the single-chip microcomputer 13, the output shaft of the motor two 77 drives the rotation of the frontmost rotating column 73, the frontmost rotating column 73 drives the rotation of the blade 74 through the transmission of the chain wheel 76 and the chain 75, the rotation of the blade 74 adjusts the mist diffusion speed, realizes the uniform distribution of humidity, better simulates the physiological environment in the body, then the motor one 56 operates, the output shaft of the motor one 56 drives the rotation of the worm 55, the worm 55 drives the synchronous rotation of the rotating shaft 53 and the magnetic rotor 52 through the meshing of the worm wheel 54, the magnetic rotor 52 generates a rotating magnetic field, the rotating magnetic field of the magnetic rotor 52 penetrates the branch plate 2 and forms magnetic coupling with the stirrer 51 in the culture dish 3, drives the circular rotation of the stirrer 51, so that the cell suspension or culture medium in the culture dish 3 forms a vortex, realizes the non-contact stirring of the culture medium, and ensures the uniform mixing of the cell suspension or culture medium, when it is needed to sample the cells in the culture dish 3, the rubber sealing plug column 14 is removed, the hand is inserted into the sampling hole to sample the cells, after sampling, the rubber sealing plug column 14 is quickly inserted into the inside of the sampling hole to reduce gas leakage.

[0029] It is worth noting that the motor 56, ultrasonic humidifier 71, motor 77, heating plate 8, temperature sensor 9, mass flow controller 11, pressure sensor 12 and humidity sensor 16 disclosed in the above embodiments, the motor 56 and the motor 77 can be selected KS-370 motor, the ultrasonic humidifier 71 can be selected HUMI300 ultrasonic humidifier, the temperature sensor 9 can be selected OHR-WS50 temperature sensor, the mass flow controller 11 can be selected GMFM-CXB-4-R4-D-A1-F1 gas flow controller, the pressure sensor 12 can be selected MPX5700AP pressure sensor, the humidity sensor 16 can be selected SHT31-DIS humidity sensor, the single-chip microcomputer control motor 56, ultrasonic humidifier 71, motor 77, heating plate 8, temperature sensor 9, mass flow controller 11, pressure sensor 12 and humidity sensor 16 work all adopt the method commonly used in the prior art.

[0030] The above only describes the embodiments of the present application, and does not limit the patent range of the present application, and any equivalent structure or equivalent process transformation, or direct or indirect application in other related technical fields by using the content of the present application specification and drawings, are also included in the patent protection range of the present application.

Claims

1. A hypoxic environment simulated cell culture device, comprising an incubator (1), wherein a support plate (2) is provided at the center of the incubator (1), heating plates (8) are respectively provided on the left and right inner walls of the incubator (1), a placement ring (4) is provided at the upper middle part of the support plate (2), and a culture dish (3) is provided inside the placement ring (4), characterized in that: It also includes stirring mechanism (5) and humidification mechanism (7); Stirring mechanism (5): it includes stirring sub (51), magnetic rotor (52) and rotating shaft (53), the middle part of the bottom wall of the incubator (1) is rotatably connected with rotating shaft (53), the upper end of rotating shaft (53) is fixedly connected with magnetic rotor (52), the inside of culture dish (3) is provided with stirring sub (51), Humidification mechanism (7): it includes ultrasonic humidifier (71), airway (72), rotating column (73) and blade (74), the left and right sides of the bottom wall of the incubator (1) are respectively provided with ultrasonic humidifier (71), the upper end of ultrasonic humidifier (71) is respectively provided with airway (72), the left and right inner walls of airway (72) are rotatably connected with evenly distributed blades (74) through rotating column (73) respectively.

2. The hypoxic environment mimicking cell culture device according to claim 1, wherein: The right side of the front end of the incubator (1) is provided with single-chip microcomputer (13), the input end of single-chip microcomputer (13) is electrically connected with external power supply, the input end of heating plate (8) and ultrasonic humidifier (71) is electrically connected with the output end of single-chip microcomputer (13).

3. The hypoxic environment mimicking cell culture device according to claim 2, wherein: The stirring mechanism (5) further comprises a worm gear (54), a worm (55) and a motor (56), the bottom wall of the incubator (1) is provided with a protective shell (6), the outer wall of the rotating shaft (53) is fixedly sleeved with the worm gear (54), the right side between the front and rear inner walls of the protective shell (6) is rotatably connected with the worm (55), the worm gear (54) is meshed with the worm (55), the front end of the incubator (1) is provided with the motor (56), the output shaft rear end of the motor (56) is fixedly connected with the front end of the worm (55), the input end of the motor (56) is electrically connected with the output end of the single-chip microcomputer (13).

4. The hypoxic environment mimicking cell culture device according to claim 2, wherein: The humidification mechanism (7) further comprises a chain (75), a sprocket (76) and a motor (77), the left side outer wall of the rotating column (73) is fixedly connected with two sprockets (76) respectively, the longitudinally adjacent and located in the same vertical plane two sprockets (76) are drivingly connected through the chain (75) between them respectively, the front side of the right end of the airway (72) is respectively provided with the motor (77), the output shaft left end of the motor (77) is respectively fixedly connected with the right end of the rotating column (73) of the frontmost end transversely adjacent, the input end of the motor (77) is electrically connected with the output end of the single-chip microcomputer (13).

5. The hypoxic environment mimicking cell culture device of claim 2, wherein: The left and right inner walls and the back wall of the incubator (1) are respectively provided with temperature sensor (9) and humidity sensor (16), the upper end of the incubator (1) is provided with pressure sensor (12), the temperature sensor (9), the pressure sensor (12) and the humidity sensor (16) are bidirectionally electrically connected with the single-chip microcomputer (13).

6. The hypoxic environment simulation cell culture device according to claim 2, wherein: The air inlet of the upper end of the incubator (1) is respectively provided with air inlet pipe (10), the middle part of the air inlet pipe (10) is respectively connected with mass flow controller (11), the mass flow controller (11) is bidirectionally electrically connected with the single-chip microcomputer (13).

7. The hypoxic environment simulation cell culture device of claim 1, wherein: The middle part of the front end of the incubator (1) is hingedly connected with cover plate (15), the inside of the sampling hole of the front end middle part of the cover plate (15) is inserted with rubber sealing plug column (14).

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