Cell is anoxia experimental facility intermittently
Technical field
The present invention relates to a kind of cell culture experiments device, more precisely a kind of cell anoxia experimental facility at intermittence.
Background technology
Sleep apnea syndrome (sleep apnea syndrome, SAS), especially obstructive sleep apnea low syndrome (obstructive sleep apnea-hyponea syndrome, OSAHS), be the chronic sleep-respiratory illness that a kind of sickness rate is higher, have a strong impact on people's physical and mental health.SAS means every night 7h sleep, breathlessness repeatedly show effect more than 30 times or sleep disordered breathing time index (respiratory impair time index, RIT, i.e. average sleep apnea number of times per hour+low ventilation number of times) >=5 times.Cause thus the hypoxemia and the hypercapnia that repeatedly occur, on body, can produce many-sided impact: as drowsiness, dizzy weak, mental act abnormal, play morning such as headache etc.With disease progression, can there is the severe complications such as pulmonary hypertension, pulmonary heart disease, respiratory insufficiency, hypertension, heart disorder.
Although SAS has been carried out to some relevant researchs, its clinical manifestation and substantially clear on the impact of internal organs both at home and abroad.But up to the present,, the pathogenesis of SAS does not still understand, is mainly in default of a kind of animal model comparatively approaching with SAS pathogenic process.So, copying the animal model close with SAS nature pathological and physiological condition, the pathogenesis to research SAS, formulates prevention and treatment measure, all has great importance.Due at present still unclear to this sick cause of disease, pathogenesis, on human body, study certain limitation, so it is particularly important that the research of relevant animal model seems, for this reason, the applicant is by having researched and developed for many years multiple intermittent hypoxia Animal feeding cabin, the name of Bing Yi Wenzhou Medical College has been declared respectively patent of invention: 1. SPF level intermittent hypoxia rat feeding compartment, and the patent No.: 200610052684.5, authorize day: 2008.7.25.; 2. intermittent hypoxia big and small mice feeding cabin of loop track type, the patent No.: 200910099827.1, authorize day: 2011.05.04.; 3. intermittent hypoxia big and small mice feeding cabin of linear track type, the patent No.: 200910099828.6, authorize day: 2010.12.08.; 4. hypoxemia Animal feeding cabin fault emergency protecting equipment, number of patent application: 201210059548.4 applyings date: 2012. 3.8.By the test to the animal model of animal for many years, the pathogenetic research of SAS is also made some progress.Yet because comprise that the mankind's biology is comprised of cell, cell is the fundamental unit of organism structure and function, organism is that the activity by cell reflects its function.Cell is a relatively independent unit, and existing his life works for the common whole life forming of other cells again, and biological generation disease is because its cell malfunction.But to the research of SAS, be to be all based upon on the animal model of intermittent hypoxia conventionally both at home and abroad at present, not yet further carry out the research of intermittent hypoxia cell.Some experimental study is to adopt animal model to do, and some is more reliable with cell experiment, more careful, and also having in addition some cell experiments is supplementing of animal.Due to intermittently anoxic experiment of cell, gas frequently flows and exchange, can take away moisture content and heat in Laboratory Module during gas flow, now keeps the constant very important of the interior humidity of Laboratory Module, temperature, to ensure that cell can normal survival and growth.And existing intermittent hypoxia Animal feeding cabin is owing to not having the environment of constant temperature, constant humidity, existing two gas, three gas cell culture incubators cannot carry out frequent exchange and humidity is controlled, therefore all do not have the research of carrying out intermittent hypoxia cell, be badly in need of intermittently anoxia experimental facility of a kind of cell of research and development.
Summary of the invention
The object of the invention is in order to overcome the deficiencies in the prior art, and a kind of cell anoxia experimental facility at intermittence that can carry out intermittent hypoxia cell research is provided.
For achieving the above object, the invention discloses intermittently anoxia experimental facility of a kind of cell, comprise cell culture incubator and Tissue Culture Dish, it is characterized in that: described experimental installation is also provided with humidity conditioner, temperature-control device, oxygen source, carbon dioxide air source, nitrogen source of the gas and principal controller, described oxygen source, carbon dioxide air source, nitrogen source of the gas is connected with cell culture incubator through corresponding gas pipe line respectively, in described gas pipe line, be equipped with by-pass valve control, humidity conditioner described in described main controller controls, temperature-control device makes in cell culture incubator in constant temperature, humidity constant state, and by-pass valve control makes oxygen content periodical change between two states of height in cell culture incubator.
This device is owing to having temperature and humidity control device, make in the situation that gas frequently flows and exchanges, ensured the temperature, the humidity environment that in casing, maintain applicable Growth of Cells, and between two states of height, change formation cycle anaerobic environment by oxygen content in by-pass valve control periodic adjustment cell culture incubator described in main controller controls, it is constant that by-pass valve control keeps gas concentration lwevel simultaneously, create the intermittently environment of anoxic, carry out the research of intermittent hypoxia cell.
As the further setting of the present invention, this cell intermittently anoxia experimental facility also has a set of cellular control unit experimental installation, comprise control group culture dish and for measuring the dissolved oxygen meter of the dissolved oxygen concentration of contrast culture ware substratum, described dissolved oxygen meter detects according to the dissolved oxygen in culture dish, described dissolved oxygen meter output is connected with principal controller, the oxygen concentration of main controller controls control group remains on 21%, and gas concentration lwevel remains on 5%.
By contrast culture ware and dissolved oxygen meter are set, at any time by detecting the dissolved oxygen in contrast culture ware substratum, and detected result is delivered in principal controller and contrasted with set(ting)value, as depart from predetermined set value, control corresponding by-pass valve control and stablize the oxygen level of air in casing, measure and can not cause interference to the Growth of Cells in Tissue Culture Dish simultaneously.
As the further setting of the present invention, carbon dioxide content constant is 5% in described main controller controls casing, oxygen content 30% and lower than 1% between change.Keep carbon dioxide content constant, regulate separately oxygen to reach and regulate oxygen content in cell culture incubator, this regulative mode is fairly simple effectively; Oxygen content 30% and lower than 1% between, this is faster for what dissolved oxygen concentration in cell culture fluid was changed, and the state of anoxic at intermittence occurs in human body close to cell more.
As the further setting of the present invention, described humidity conditioner comprises the water storage tray that is arranged on described cabinets cavity bottom, described water storage tray below is provided with well heater, between described water storage tray top and Tissue Culture Dish, be provided with dividing plate by described water storage tray and Tissue Culture Dish place spatial separation, described dividing plate is provided with transom, the position of corresponding transom is slidably provided with for regulating the flashboard of the ventilative area of transom, and being provided with flashboard driving mechanism, described principal controller drives flashboard to slide by controlling flashboard driving mechanism.That the humidity conditioner of said structure has is simple in structure, it is convenient to control, effective advantage.
Below in conjunction with the drawings and specific embodiments, the present invention is further illustrated.
Accompanying drawing explanation
Accompanying drawing 1 is specific embodiment of the invention surface structure block diagram;
Accompanying drawing 2 is accompanying drawing 1 A-A sectional view;
Accompanying drawing 3 is specific embodiment of the invention STRUCTURE DECOMPOSITION schematic diagram;
Accompanying drawing 4 is another STRUCTURE DECOMPOSITION schematic diagram of the specific embodiment of the invention.
Embodiment
As Fig. 1, shown in 2, cell intermittently anoxia experimental facility comprises a cell culture incubator consisting of airtight casing 1, in cell culture incubator 1, be provided with Tissue Culture Dish 8 and temperature and humidity control device, in cell culture incubator 1 outside, be provided with oxygen source, carbon dioxide air source, nitrogen source of the gas and principal controller 2 thereof, oxygen source, carbon dioxide air source and nitrogen source of the gas can adopt medical high-pure gas steel cylinder, described oxygen source, carbon dioxide air source, nitrogen source of the gas is connected with cell culture incubator 1 through corresponding gas pipe line 3 respectively, gas pipe line 3 can be selected food grade high-pressure air pipe, in described every gas pipe line 3, be equipped with by-pass valve control (not shown), by-pass valve control is to select magnetic valve to be advisable, convenient control.Described principal controller 2 be take the automatic control system that micro-chip is that core and peripheral components form and is formed, described principal controller 2 is by controlling the opening and closing of described by-pass valve control, reach the flow of controlling various gases, and then reach the object that in periodic adjustment cell culture incubator 1, oxygen content changes between two states of height.As shown in Figure 3,4, humidity conditioner comprises the water storage tray 4 that is arranged on described cell culture incubator 1 inner chamber bottom, and described water storage tray 4 belows are provided with well heater 5, and well heater 5 can be selected ptc heater part.Between described water storage tray 4 tops and Tissue Culture Dish 8, be provided with dividing plate 6 by described water storage tray 4 and Tissue Culture Dish 8 place spatial separation, described dividing plate 6 is provided with transom 601, the position of corresponding transom 601 is slidably provided with for regulating the flashboard 7 of transom 601 ventilative areas, and is provided with flashboard 7 driving mechanisms.Wherein, dividing plate 6 can be bilevel sandwich structure, and flashboard 7 is slidably disposed in interlayer, and transom 601 can arrange a plurality of as required, and transom 601 shapes are not limit, but are advisable with regular shape, convenient processing.It in this specific embodiment, is rectangle, left and right respectively arranges one, being conducive to interior humidity Quick uniform regulates, two transoms 601 are equipped with a flashboard 7, the side that described two flashboards 7 are relative is respectively extended one section of tooth bar 701 in opposite directions, tooth bar 701 directions parallel with slip direction, between described two tooth bars 701, be provided with driving gear 9 and tooth bar 701 engagements, driving gear 9 is fixed in the rotating shaft of micromachine 10 and forms flashboard 7 driving mechanisms, described principal controller 2 is connected with micromachine 10 to be controlled the rotating speed of micromachine 10 and turns to, the direction that described principal controller 2 drives flashboard 7 to slide and stop and sliding by controlling flashboard 7 driving mechanisms, regulate the flashboard 7 of transom 601 ventilative areas, the inlet of the water vapour that control is evaporated from water storage tray 4, reach the object of the humidity in adjusting and stable culturing room.Described principal controller 2 controls that the interior carbon dioxide content of cell culture incubator 1 is constant is being advisable with 5%, oxygen content 30% and lower than 1% between change, the duration in each cycle was advisable with 10 ~ 20 minutes.In order to stablize two oxygen level under state, in this specific embodiment, in cell culture incubator 1 except being provided with Tissue Culture Dish 8, also be provided with other a set of cellular control unit experimental installation, the difference of cellular control unit experimental installation and experimental group cell experiment device is only that the oxygen concentration of control group is to remain on 21% always.Control group experimental installation is provided with contrast culture ware 11 and for measuring the dissolved oxygen meter of the dissolved oxygen concentration of contrast culture ware 11 substratum.Tissue Culture Dish 8 and contrast culture ware 11 be 96 orifice plates preferably, the oxygen sensor 12 of described dissolved oxygen meter inspection contacts with substratum in contrast culture ware 11, to detect the dissolved oxygen concentration in contrast culture ware 11, described dissolved oxygen meter output is connected with principal controller 2, by the dissolved oxygen concentration detecting in contrast culture ware 11 substratum, grasp the dissolved oxygen concentration in Tissue Culture Dish 8 at any time, and detected result is delivered in principal controller 2 with the set(ting)value of oxygen level under two states in principal controller 2 and contrasted, as depart from predetermined set value, control corresponding by-pass valve control and stablize the oxygen level of air in casing, the set(ting)value of oxygen level can be stored in the internal memory of SCM system, the comparison of numerical value and control process and mode can realize by the control algolithm program of micro-chip inside, repeat no more herein.The present invention grasps the dissolved oxygen concentration in Tissue Culture Dish 8 by the dissolved oxygen concentration detecting in control group and experimental group Tissue Culture Dish 8 substratum, can reduce the interference while measuring, the Growth of Cells in Tissue Culture Dish 8 being caused, and improves the accuracy of experiment.The more important thing is the humidity in Laboratory Module while controlling gas flow, to ensure the environment of cell normal growth.