CN105062889B - A kind of cell volatile matter collects incubator - Google Patents

A kind of cell volatile matter collects incubator Download PDF

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Publication number
CN105062889B
CN105062889B CN201510459096.2A CN201510459096A CN105062889B CN 105062889 B CN105062889 B CN 105062889B CN 201510459096 A CN201510459096 A CN 201510459096A CN 105062889 B CN105062889 B CN 105062889B
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Prior art keywords
beaker
bend pipe
cell
dish
culture dish
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CN105062889A (en
Inventor
张卓
史策
李丹丹
赵艳秋
魏红
周晋
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Harbin Medical University
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Harbin Medical University
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00Constructional details, e.g. recesses, hinges
    • C12M23/02Form or structure of the vessel
    • C12M23/08Flask, bottle or test tube
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00Constructional details, e.g. recesses, hinges
    • C12M23/02Form or structure of the vessel
    • C12M23/04Flat or tray type, drawers
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00Constructional details, e.g. recesses, hinges
    • C12M23/02Form or structure of the vessel
    • C12M23/10Petri dish
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00Constructional details, e.g. recesses, hinges
    • C12M23/36Means for collection or storage of gas; Gas holders
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M33/00Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
    • C12M33/04Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles

Abstract

The present invention relates to a kind of cell volatile matter for disease biomarker research to collect incubator, and it includes:One culture dish, the inside of the culture dish are loaded with liquid to be detected;The inner of one long bend pipe and a short bend pipe is placed in the culture dish;One beaker back-off is in the liquid of the culture dish, and the inner of the long bend pipe is higher than the liquid surface and positioned at the top of the beaker, and the inner of the short bend pipe is less than the liquid surface and positioned at the bottom of the beaker;One crystallising dish cover is buckled on the beaker, and the outer end of the long bend pipe and short bend pipe stretches out in the outside of the crystallising dish.The incubator is the device of easy, non-seal, it not only ensure that intercellular gas exchanges inside and outside beaker, and cytoactive is influenceed smaller, the cell volatile matter for making to be collected into more has Research Significance, therefore it is more beneficial for probing into the metabolic mechanism of disease, beneficial to the preliminary examination and early diagnosis of the diseases such as tumour.

Description

A kind of cell volatile matter collects incubator
Technical field
The present invention relates to a kind of cell culture apparatus, more particularly to one kind is used to collect cell volatility under non-closed environment The cell culture apparatus of biomarker.
Background technology
At present, the diagnosis of tumour needs the invasive inspection such as histopathology, aspiration biopsy more, but these check that time-histories is longer, especially It is it is traumatic operate made troubles to patient and pain.Therefore, urgently exploitation it is a kind of simple, effectively, it is economical, conveniently, it is noninvasive Diagnostic techniques be used for disease early screening and clinical diagnosis.In recent years, people at a variety of aspects to being waved caused by solid tumor Hair property organic compound is studied, including the blood of tumor patient, urine, expiratory air, tumor tissues and tumour cell, Attempt to be used as disease early screening and the new method diagnosed by finding volatility label.Human body is one comprising a variety of dirty The complexity individual of device, there is the metabolism that a variety of enzymes and metabolic system participate in body, and the pathophysiological process of disease;And because Influenceed by factors such as diet, the volatility label for studying human body fluid and expiratory air is relatively complicated.Therefore, in cellular level The volatility label of upper analysis disease will be helpful to further probe into may metabolic mechanism caused by disease.
At present, existing document discloses a kind of cell culture for being used to detect cell VOC and detection dress Put, cultivated using glass blake bottle, but the device is seal, and such a closed environment is unfavorable for cell and carries out gas friendship Change, it is impossible to suitable culture environment is provided for cell, easily causes the anoxia state of cell, causes Apoptosis influenceing cell just Often it is metabolized, the organic compound that cell volatilizes under such a environment is most likely not to be metabolized output under cell normal condition to be detected Material, be unfavorable for tumor marker really cutting analyse, and its not under such a culture environment cell activity and state enter Row analysis, therefore the real reliability of experimental result needs further to be investigated.
Separately there is document to also disclose that a kind of cell culture apparatus, for detecting the VOC of cell, but should Device construction is complicated, and it need to be joined directly together with the gas tank containing 95%CO2 and 5%O2, and need catalyst, hydrocarbon to catch Storage, heat distribution formula mass flow controller, sterile filters, heating water bath circuit, magnet, electronic control valve, dilution pipeline etc. A variety of accessories, therefore operation is relatively cumbersome, expense is also of a relatively high.
The content of the invention
In order to solve the above technical problems, it is an object of the invention to:A kind of cell volatile matter is provided and collects incubator, the training The device that device is easy, non-seal is supported, not only ensure that the gas exchanges of cell inside and outside beaker, and to cytoactive shadow Sound is smaller, and the cell volatile matter for making to be collected into more has Research Significance, therefore is more beneficial for probing into the metabolic mechanism of disease, beneficial to swollen The preliminary examination and early diagnosis of the diseases such as knurl.
To achieve the above object, the technical solution adopted in the present invention is:A kind of cell volatile matter collects incubator, and it is wrapped Include:
One culture dish, liquid to be detected is loaded with inside the culture dish;
The inner of one long bend pipe and a short bend pipe, the long bend pipe and short bend pipe is placed in the culture dish;
One beaker, the beaker back-off is in the liquid of the culture dish and surrounds the inner of the long bend pipe and short bend pipe, wherein, The inner of the long bend pipe is located at the top of the beaker and is higher than the liquid surface, and the inner of the short bend pipe is located at the bottom of the beaker And it is less than the liquid surface;
One crystallising dish, the crystallising dish cover are buckled on the beaker, and the outer end of the long bend pipe and short bend pipe, which is bent, stretches out in this The outside of crystallising dish.
Described cell volatile matter collects incubator, opening the opening less than the culture dish top of the crystallising dish container bottom Mouthful.
Described cell volatile matter collects incubator, the liquid level in beaker after the beaker back-off and the liquid in the culture dish Face keeps concordant.
Described cell volatile matter collects incubator, and the liquid to be detected is cell to be detected and culture medium.
Described cell volatile matter collects incubator, and the outer end of the long bend pipe connects a collector by a silicone tube, should Collector is provided with a knob with the silicone tube junction, for gas collection and maintains the pressure balance inside and outside beaker.
Described cell volatile matter collects incubator, silica gel pipe range 3cm, and the collector is 0.5L Storage Time in Gas Collecting Bag.
Described cell volatile matter collects incubator, and the specification of the culture dish is 90mm, and the specification of the beaker is 250mL, The specification of the crystallising dish is 125mm.
Described cell volatile matter collects incubator, and the diameter of the beaker is less than the diameter of culture dish.
Described cell volatile matter collects incubator, and the height of the beaker is higher than the height of the culture dish.
Described cell volatile matter collects incubator, and the diameter of the crystallising dish is more than the diameter of the culture dish.
Compared with prior art, using above-mentioned technical proposal the advantage of the invention is that:Instant invention overcomes existing skill The closed environment of cell culture and the shortcomings of complex operation, accessory redundancy in art, there is provided a kind of easy, non-seal, Low-cost device, the gas exchanges of cell inside and outside beaker are ensure that, relatively normal culture environment is provided for cell, led to Overflow-type cell instrument detects the apoptosis of cell, as a result shows that cell is active up to 92.18% after being cultivated 24 hours in this device ± 1.26%.
Brief description of the drawings
Fig. 1 is schematic structural view of the invention.
Description of reference numerals:1- culture dishes;2- testing liquids;The long bend pipes of 3-;The short bend pipes of 4-;5- beakers;6- crystallising dish;7- Silicone tube;8- collectors;81- knobs.
Embodiment
The present invention is further described with reference to specific embodiments and the drawings, advantages of the present invention and feature will be with Description and it is apparent.
As shown in figure 1, collecting the three-dimensional assembling schematic diagram of incubator for the cell volatile matter of the present invention, it includes:
One culture dish 1, liquid to be detected 2 is loaded with inside the culture dish 1;Wherein, in the case study on implementation of the present invention, treat Detection liquid 2 includes cell to be detected and culture medium, can also need to add corresponding medicine because of experimental design, the culture dish 1 Specification is 90mm, and a long 3 and one short bend pipe 4 of bend pipe is put into the culture dish 1, and the inner of the long bend pipe 3 and short bend pipe 4 is put In the culture dish 1.
Taking the back-off of beaker 5 that a specification is 250ml again, the liquid level in the beaker 5 is with being somebody's turn to do in the liquid 2 of the culture dish 1 Liquid level in culture dish 1 will ensure concordantly.Wherein, the diameter of the beaker 5 is less than the diameter of the culture dish 1, but the burning simultaneously In the inner of the long bend pipe 3 and short bend pipe 4 is enclosed in by cup 5, the inner bottom part ring in the beaker 5 of the long bend pipe 3 around Bending, and make its bottom upon bending higher than the surface of liquid 2 and extend always to the top of the beaker 5, long bend pipe 3 is used to receive Collect gas caused by beaker inner cell, the inner of the short bend pipe 4 is located at the bottom of the beaker 5 and is less than the surface of liquid 2, short curved Pipe 4 is used to draw the experiment that testing liquid carries out next step at the end of testing.
The crystallising dish 6 that a specification is 125mm is buckled on the beaker 5 again, because the height of beaker 5 is higher than the culture dish 1 height, and the diameter of crystallising dish 6 is more than the diameter of culture dish 1, therefore when the cover of crystallising dish 6 is buckled on beaker 5, container knot The opening of the brilliant bottom of ware 6 has a certain distance with the bottom of culture dish 1, therefore, between culture dish 1 and crystallising dish 6 forms a circle The space of ring-type, but the horizontal plane where the opening of the container bottom of crystallising dish 6 will be less than where the opening on the top of culture dish 1 Horizontal plane, ensure that other materials are not polluted the testing liquid 2 in the culture dish 1 in by outside air, meanwhile, the long bend pipe 3 and then bending stretches out in the outside of the crystallising dish 6 outer end of short bend pipe 4.
The outer end of the long bend pipe 3 connects a collector 8, the collector 8 and the silicone tube 7 by the silicone tube 7 of a long 3cm Junction is provided with knob 81, for gas collection and maintains the pressure balance inside and outside beaker.In the case study on implementation of the present invention, collect Device 8 using 0.5L Storage Time in Gas Collecting Bag.
Therefore to be learnt by above content, the upper opening of culture dish 1 is unclosed state in apparatus of the present invention, meanwhile, crystallization The diameter of ware 6 is more than the diameter of culture dish 1, and the bottom of the opening of the bottom of container crystallising dish 6 and culture dish 1 have it is certain away from From therefore, the circular space of formation one between culture dish 1 and crystallising dish 6, therefore, the cell in culture dish 1 is in non-closed loop Gas exchanges can be carried out with the cell in beaker 5 under border, caused gas is placed in the top of beaker 5, and long bend pipe 3 passes through collector 8 collect gas, and the volatile materials after collection further carries out SPME and GC/MS Analysis etc., and other are metabolized Group credit analysis, short bend pipe 4 are used to extract cell to further testing inspection.Such as Apoptosis, PCR.
It is meanwhile scarce instant invention overcomes the closed environment of cell culture in the prior art and complex operation, accessory redundancy etc. Point, there is provided a kind of easy, non-seal, low-cost device, the gas exchanges of cell inside and outside beaker 5 are ensure that, It is smaller on cytoactive influence, relatively normal culture environment is provided for cell, passes through withering for flow cytomery cell Die, as a result show that cell is active up to 92.18% ± 1.26% after being cultivated 24 hours in this device;Therefore volatilize cell Organic compound it is more genuine and believable, therefore be more beneficial for probing into the metabolic mechanism of disease, preliminary examination beneficial to disease and Early diagnosis.
Explanation and embodiment above is only exemplary, does not form any restrictions to the scope of the present invention.This area Technical staff should be understood that without departing from the spirit and scope of the invention can to the details of technical solution of the present invention and Form is modified or replaced, but these modifications and replacement are each fallen within protection scope of the present invention.

Claims (6)

1. a kind of cell volatile matter collects incubator, it is characterised in that it includes:
One culture dish, liquid to be detected is loaded with inside the culture dish;
The inner of one long bend pipe and a short bend pipe, the long bend pipe and short bend pipe is placed in the culture dish;
One beaker, the beaker back-off is in the liquid of the culture dish and surrounds the inner of the long bend pipe and short bend pipe, wherein, the length The inner of bend pipe is located at the top of the beaker and is higher than the liquid surface, and the inner of the short bend pipe is located at the bottom of the beaker and low In the liquid surface;
One crystallising dish, the diameter of the crystallising dish are more than the diameter of the culture dish, and the height of the beaker is higher than the height of the culture dish, The crystallising dish cover is buckled on the beaker, and the opening of the crystallising dish container bottom is less than the opening on the culture dish top, the long bend pipe The outside for stretching out in the crystallising dish is bent with the outer end of short bend pipe;
The outer end of the long bend pipe connects a collector by a silicone tube, and the collector is provided with rotation with the silicone tube junction Button, for gas collection and the pressure balance inside and outside beaker is maintained, the short bend pipe is used to extract cell.
2. cell volatile matter according to claim 1 collects incubator, it is characterised in that in the beaker after the beaker back-off Liquid level it is concordant with the liquid level holding in the culture dish.
3. cell volatile matter according to claim 1 collects incubator, it is characterised in that the liquid to be detected is to be detected Cell and culture medium.
4. cell volatile matter according to claim 1 collects incubator, it is characterised in that silica gel pipe range 3cm, the collection Device is 0.5L Storage Time in Gas Collecting Bag.
5. cell volatile matter according to claim 1 collects incubator, it is characterised in that the specification of the culture dish is 90mm, the specification of the beaker is 250mL, and the specification of the crystallising dish is 125mm.
6. cell volatile matter according to claim 1 collects incubator, it is characterised in that the diameter of the beaker is less than culture The diameter of ware.
CN201510459096.2A 2015-07-30 2015-07-30 A kind of cell volatile matter collects incubator Active CN105062889B (en)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102089420A (en) * 2008-07-08 2011-06-08 威尔森沃尔夫制造公司 Improved gas permeable cell culture device and method of use
CN202886148U (en) * 2012-08-09 2013-04-17 山东省产品质量监督检验研究院 Sampling collector of organic volatile matters in product/sample
CN203613193U (en) * 2013-10-23 2014-05-28 上海交通大学医学院附属仁济医院 Dynamic detection device for metabolism and respiration of cells

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5612188A (en) * 1991-11-25 1997-03-18 Cornell Research Foundation, Inc. Automated, multicompartmental cell culture system

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102089420A (en) * 2008-07-08 2011-06-08 威尔森沃尔夫制造公司 Improved gas permeable cell culture device and method of use
CN202886148U (en) * 2012-08-09 2013-04-17 山东省产品质量监督检验研究院 Sampling collector of organic volatile matters in product/sample
CN203613193U (en) * 2013-10-23 2014-05-28 上海交通大学医学院附属仁济医院 Dynamic detection device for metabolism and respiration of cells

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