CN203530332U - Cell culture device capable of monitoring cell culture liquid environment - Google Patents
Cell culture device capable of monitoring cell culture liquid environment Download PDFInfo
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- CN203530332U CN203530332U CN201320697500.6U CN201320697500U CN203530332U CN 203530332 U CN203530332 U CN 203530332U CN 201320697500 U CN201320697500 U CN 201320697500U CN 203530332 U CN203530332 U CN 203530332U
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- cell culture
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- 238000004113 cell culture Methods 0.000 title claims abstract description 73
- 238000012544 monitoring process Methods 0.000 title claims abstract description 35
- 239000007788 liquid Substances 0.000 title claims abstract description 31
- 239000000463 material Substances 0.000 claims description 13
- 229920001903 high density polyethylene Polymers 0.000 claims description 10
- 239000004700 high-density polyethylene Substances 0.000 claims description 10
- 239000004793 Polystyrene Substances 0.000 claims description 4
- 229920002223 polystyrene Polymers 0.000 claims description 4
- 238000000034 method Methods 0.000 abstract description 14
- 230000008569 process Effects 0.000 abstract description 12
- 239000000126 substance Substances 0.000 abstract description 10
- 238000007789 sealing Methods 0.000 abstract 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 28
- 239000001301 oxygen Substances 0.000 description 28
- 229910052760 oxygen Inorganic materials 0.000 description 28
- 235000015097 nutrients Nutrition 0.000 description 21
- 210000004027 cell Anatomy 0.000 description 16
- 230000010261 cell growth Effects 0.000 description 6
- 238000012258 culturing Methods 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 5
- 206010021143 Hypoxia Diseases 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 239000007789 gas Substances 0.000 description 4
- 230000001146 hypoxic effect Effects 0.000 description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 3
- 239000012930 cell culture fluid Substances 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- GQPLMRYTRLFLPF-UHFFFAOYSA-N Nitrous Oxide Chemical compound [O-][N+]#N GQPLMRYTRLFLPF-UHFFFAOYSA-N 0.000 description 2
- 230000001464 adherent effect Effects 0.000 description 2
- 239000006143 cell culture medium Substances 0.000 description 2
- 230000019522 cellular metabolic process Effects 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 239000004310 lactic acid Substances 0.000 description 2
- 235000014655 lactic acid Nutrition 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 150000004767 nitrides Chemical class 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 206010058490 Hyperoxia Diseases 0.000 description 1
- 206010044074 Torticollis Diseases 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000006706 cellular oxygen consumption Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000012611 container material Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 235000012041 food component Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000000222 hyperoxic effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 208000018197 inherited torticollis Diseases 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000012806 monitoring device Methods 0.000 description 1
- 230000036284 oxygen consumption Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
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Abstract
The utility model discloses a cell culture device capable of monitoring the cell culture liquid environment. The cell culture device comprises a monitoring meter, a cell culture bottle, sensor electrodes and sealing rings, wherein the cell culture bottle takes the shape of a cuboid, and a bottle neck is upwards obliquely arranged at one end of the cell culture bottle; at least two electrode sockets are arranged on the cell culture bottle, and each electrode socket takes the shape of a cylindrical bore extending from a bottle body to the interior of the bottle body; the sensor electrodes are inserted into the electrode sockets through the sealing rings sleeved on the sensor electrodes, and the monitoring meter is connected with the sensor electrodes through conducting wires. The cell culture device has the benefits that the real-time concentration of a particular substance in cell culture liquid can be monitored in the cell or tissue culture process, and the influences of the particular culture liquid environment on cells or tissues can be researched more accurately and dynamically.
Description
Technical field
The utility model relates to cellular engineering cells in vitro culture apparatus, is specifically related to a kind of cell culture apparatus of monitoring cell cultures liquid environment.
Background technology
Cells in vitro culture technique is that mammalian cell is inoculated in the liquid culture medium in cell culture container, wherein contain Growth of Cells and breed necessary multiple nutritional components, and place it in the cell culture incubator that specified temp, humidity, atmosphere surrounding can be set and hatch cultivation, make it growth, propagation.In scientific research, often need to study the impact of certain predetermined substance process such as cell growth, division, propagation under certain concentration in cell culture fluid.These predetermined substances comprise that dissolved oxygen in cell culture fluid, pH, supercarbonate, glucose, mineral ion, products of cellular metabolism are as lactic acid, nitride, nitrogen protoxide etc.The change in concentration of predetermined substance in cannot Real-Time Monitoring cell cultures liquid in process of experimental in the past, while especially needing long-time (> 24h) to cultivate, be difficult to the complete metabolic process of certain predetermined substance under dynamically recording, therefore cannot obtain accurately experimental result all sidedly.If by increasing cell cultures number of times and a plurality of nutrient solution sampling time point being set and detecting number of times and obtain dynamic data, will make again experimentation too loaded down with trivial details, and need to drop into a large amount of man power and materials, also easily cause the pollution of cell cultures simultaneously.In addition, when carrying out anoxic or hyperoxia cell cultures, open to cultivate chamber door and gather in cell culture fluid process and will cause the variation of incubator atmosphere surrounding, and need time balance again, therefore, increase the accuracy that sampling number must affect experimental result.
Utility model content
The purpose of this utility model is exactly for above-mentioned defect of the prior art, provide a kind of can Real-Time Monitoring cell cultures state, do not affect the cell culture apparatus of cell culture environment.
To achieve these goals, the technical scheme that the utility model provides is: a kind of cell culture apparatus of monitoring cell cultures liquid environment, includes monitoring meter 1, Tissue Culture Flask 5, sensor electrode 2 and sealing-ring 8; Described Tissue Culture Flask 5 is cuboid; One end of described Tissue Culture Flask 5 is inclined upwardly and is provided with bottleneck 6; On described bottleneck 6, be provided with bottle cap 7; On described Tissue Culture Flask 5, be provided with at least two electrode sockets 4; Described electrode socket 4 be shaped as by bottle to bottle chamber the inner cylinder hole shape extending; The aperture of described electrode socket 4 is greater than the aperture, lower end of sensor electrode 2; Described sensor electrode 2 is plugged in electrode socket 4 by the sealing-ring 8 being sheathed on sensor electrode 2; Described sealing-ring 8 upper ends are ring-type, and lower end is cylindrical; The cylindrical aperture of described sealing-ring 8 lower ends is greater than the aperture of sensor electrode 2 lower ends and is less than the aperture of electrode socket 4; Described monitoring meter 1 is connected with sensor electrode 2 by wire 3.
Further, above-mentioned a kind of cell culture apparatus of monitoring cell cultures liquid environment, 7 of described bottleneck 6 and bottle caps are for being threaded.
Further, above-mentioned a kind of cell culture apparatus of monitoring cell cultures liquid environment, described electrode socket 4 is provided with three.
Further, above-mentioned a kind of cell culture apparatus of monitoring cell cultures liquid environment, the bottom of described electrode socket 4 is 1cm apart from Tissue Culture Flask 5 bottoms.
Further, above-mentioned a kind of cell culture apparatus of monitoring cell cultures liquid environment, the bottle material of described Tissue Culture Flask 5 is polystyrene; The material of described bottle cap 7 is high density polyethylene(HDPE); The material of described sealing-ring 8 is rubber.
Tissue Culture Flask of the present utility model, its shape, size and making material are all similar to traditional Tissue Culture Flask, specifically rectangle, torticollis, polystyrene (Polystrene) bottle, high density polyethylene(HDPE) (High Density Polyethylene) seal bottle cap.Difference is: culturing bottle has some invaginatings, the columniform electrode socket of inwall away from one end of bottleneck, socket inwall extends downward apart from bottle end 1cm place, socket entrance has rubber seal, when not using, the electrode socket stopper 9 of available high-density polyethylene material covers tightly, and guarantees sealed air-tight.
Be used for monitoring while cultivating liquid content of predetermined substance, first should guarantee that in Tissue Culture Flask, nutrient solution liquid level is at least 8mm, electrode socket stopper is pulled out and confirmed that sealing-ring is still in place simultaneously again, afterwards sensor electrode be inserted in electrode socket and be fully immersed in nutrient solution.Tissue Culture Flask is positioned in cell culture incubator, with cable, sensor electrode is connected with the monitoring meter outside cell culture incubator, can be used for monitoring the real-time concentration of cultivating certain predetermined substance in liquid in cell or tissue culturing process, thereby more accurately, dynamically study the impact of certain predetermined substance process such as cell growth, division, propagation under certain concentration.
The beneficial effects of the utility model are: the cell culture apparatus that the utility model provides, the real-time concentration of certain predetermined substance in cell cultures liquid can be monitored in cell or tissue culturing process, the impact of specific cultivation liquid environment on cell or tissue can be more accurately, dynamically studied.According to different experiments demand, the electrode sensor monitoring nutrient solution content of predetermined substance of capable of choosing multiple type changes, as dissolved oxygen electrode, pH electrode, carbon dioxide electrode, glucose electrode, lactic acid electrode, nitride electrode, nitrogen protoxide electrode, ion electrode etc.; Also can select integrated combined electrode or insert the Real-Time Monitoring that multiple sensors electrode is realized multi-path parameter simultaneously, during customization electrode sensor, requiring electrode shell diameter specifications and culturing bottle electrode socket internal diameter to match.
The cell culture apparatus that the utility model provides is not limited only to Tissue Culture Flask, also can be the containers such as Tissue Culture Dish, plate, and container material quality can be glass or macromolecular material.
Accompanying drawing explanation
Fig. 1 is integrated connection schematic diagram of monitoring the cell culture apparatus of cell cultures liquid environment of the present utility model.
Fig. 2 is that the utility model can be monitored in the cell culture apparatus of cell cultures liquid environment, the use state vertical section connection diagram of Tissue Culture Flask.
Fig. 3 is that the utility model can be monitored in the cell culture apparatus of cell cultures liquid environment, the non-working condition vertical section connection diagram of Tissue Culture Flask.
Fig. 4 is that the utility model can be monitored in the cell culture apparatus of cell cultures liquid environment, the cross sectional representation of Tissue Culture Flask.
Fig. 5 is the schematic diagram that the utility model can be monitored sensor electrode in the cell culture apparatus of cell cultures liquid environment.
Wherein, 1 is monitoring meter, and 2 is sensor electrode, and 3 is wire, and 4 is electrode socket, and 5 is Tissue Culture Flask, and 6 is bottleneck, and 7 is bottle cap, and 8 is sealing-ring, and 9 is electrode socket stopper.
Embodiment
embodiment 1:
Monitor a cell culture apparatus for cell cultures liquid environment, include monitoring meter 1, Tissue Culture Flask 5, sensor electrode 2 and sealing-ring 8; Tissue Culture Flask 5 is cuboid, and its bottle material is polystyrene; One end of Tissue Culture Flask 5 is inclined upwardly and is provided with bottleneck 6; On bottleneck 6, be provided with bottle cap 7, the two is threaded connection, and the material of bottle cap 7 is high density polyethylene(HDPE); On Tissue Culture Flask 5, be provided with three electrode sockets 4; Electrode socket 4 be shaped as by bottle to bottle chamber the inner cylinder hole shape extending, the bottom of electrode socket 4 is 1cm apart from Tissue Culture Flask 5 bottoms; The aperture of electrode socket 4 is greater than the aperture, lower end of sensor electrode 2; Sensor electrode 2 is plugged in electrode socket 4 by the sealing-ring 8 being sheathed on sensor electrode 2; Sealing-ring 8 upper ends are ring-type, and lower end is cylindrical, and the material of sealing-ring 8 is rubber; The cylindrical aperture of sealing-ring 8 lower ends is greater than the aperture of sensor electrode 2 lower ends and is less than the aperture of electrode socket 4; Monitoring meter 1 is connected with sensor electrode 2 by wire 3.
specific operation process:
the content of nutrient solution dissolved oxygen in monitoring hypoxic cell culturing process:
The ordinary method that hypoxic cell is cultivated is to control in three gas cell culture incubators in atmosphere surrounding the concentration of oxygen in anoxic level, and the concentration of case inner cell nutrient solution dissolved oxygen also will be in anoxic level, thereby realizes the process that analog cell anoxic is cultivated.Because cell is immersed in liquid culture medium, cultivate, Real-Time Monitoring and guarantee nutrient solution dissolved oxygen concentration constant be the key of setting up stable hypoxic cell culture experiment platform.Common cell culture medium comprises MEM, DMEM, RPMI-1640,199 cell culture medium series etc., their composition there are differences, and the level pad of preparation substratum is also different, thereby cause the dissolved oxygen concentration of substratum under same oxygen partial pressure conditions different.In addition, the variation of the quantity variance of the interior viable cell of cell culture container and the nutrient solution potential of hydrogen causing at different cultivation stage cell metabolism products, also can cause nutrient solution dissolved oxygen concentration and incubator oxygen partial pressure disproportionate, especially with oxygen concentration in incubator lower than more remarkable after 10%.In order dynamically to observe nutrient solution dissolved oxygen concentration, will repeatedly open chamber door collection nutrient solution and carry out dissolved oxygen detection.And in the process that hypoxic cell is cultivated, frequently open the rise that incubator goalkeeper causes incubator oxygen concentration, and need balance 15～30min could again reach the anoxic level of setting, this must affect repeatability and the accuracy of experimental result.Therefore, need to set up and a kind ofly guarantee that cell culture environment is constant, reduce pollution of cell culture, laborsaving simple nutrient solution dissolved oxygen concentration real-time monitoring device as far as possible.
The specific experiment process of applying this device is as follows:
experiment 1:it is 5% that three gas cell culture incubators arrange culture environment oxygen concn, the Tissue Culture Flask of inoculating cell is placed in cell culture incubator, nutrient solution liquid level 1cm, tighten after bottle cap and circle round again and within one week, be beneficial to a bottle inner air and outer air circulation, after 2h, treat the abundant adherent growth of cell, pull out 1 electrode socket stopper, determine that sealing-ring is still in place simultaneously, again dissolved oxygen electrode be inserted in electrode socket and be fully immersed in nutrient solution, electrode cable is connected with the dissolved oxygen monitoring meter outside cell culture incubator, can monitor for a long time the real-time concentration of dissolved oxygen in nutrient solution, thereby more accurately, dynamically study the impact of anaerobic environment cell growth propagation.In cell cultivation process, if nutrient solution dissolved oxygen concentration changes, can feedback three gas incubator oxygen partial pressure are regulated, thereby realize the constant of nutrient solution dissolved oxygen concentration.
experiment 2:it is 5% that three gas cell culture incubators arrange culture environment oxygen concn, the Tissue Culture Flask of inoculating cell is placed in cell culture incubator, nutrient solution liquid level 1cm, tighten after bottle cap and circle round again and within one week, be beneficial to a bottle inner air and outer air circulation, after 2h, treat the abundant adherent growth of cell, pull out 1 electrode socket stopper, determine that sealing-ring is still in place simultaneously, again dissolved oxygen electrode be inserted in electrode socket and be fully immersed in nutrient solution, again screw bottle cap to stop the circulation of culturing bottle inner air and outer air, electrode cable is connected with the dissolved oxygen monitoring meter outside cell culture incubator, can monitor for a long time the real-time concentration of dissolved oxygen in nutrient solution.Cell carries out 72h cultivation (not changing liquid midway) continuously, the concentration of dissolved oxygen in dynamic monitoring nutrient solution, for study dissolved oxygen concentration and cell oxygen consumption rate, oxygen-consumption and and cell survival quantity between relation.
Finally it should be noted that: the foregoing is only preferred embodiment of the present utility model, be not limited to the utility model, although the utility model is had been described in detail with reference to previous embodiment, for a person skilled in the art, its technical scheme that still can record aforementioned each embodiment is modified, or part technical characterictic is wherein equal to replacement.All within spirit of the present utility model and principle, any modification of doing, be equal to replacement, improvement etc., within all should being included in protection domain of the present utility model.
Claims (5)
1. can monitor a cell culture apparatus for cell cultures liquid environment, it is characterized in that, include monitoring meter, Tissue Culture Flask, sensor electrode and sealing-ring; Described Tissue Culture Flask is cuboid; One end of described Tissue Culture Flask is inclined upwardly and is provided with bottleneck; On described bottleneck, be provided with bottle cap; On described Tissue Culture Flask, be provided with at least two electrode sockets; Described electrode socket be shaped as by bottle to bottle chamber the inner cylinder hole shape extending; The aperture of described electrode socket is greater than the aperture, lower end of sensor electrode; Described sensor electrode is plugged in electrode socket by the sealing-ring being sheathed on sensor electrode; Described sealing-ring upper end is ring-type, and lower end is cylindrical; The cylindrical aperture of described sealing-ring lower end is greater than the aperture of sensor electrode lower end and is less than the aperture of electrode socket; Described monitoring meter is connected with sensor electrode by wire.
2. a kind of cell culture apparatus of monitoring cell cultures liquid environment according to claim 1, is characterized in that, between the bottleneck of described Tissue Culture Flask and bottle cap for being threaded.
3. a kind of cell culture apparatus of monitoring cell cultures liquid environment according to claim 1, is characterized in that, described electrode socket is provided with three.
4. a kind of cell culture apparatus of monitoring cell cultures liquid environment according to claim 1, is characterized in that, the bottom of described electrode socket is 1cm apart from Tissue Culture Flask bottom.
5. a kind of cell culture apparatus of monitoring cell cultures liquid environment according to claim 1, is characterized in that, the bottle material of described Tissue Culture Flask is polystyrene; The material of described bottle cap is high density polyethylene(HDPE); The material of described sealing-ring is rubber.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201320697500.6U CN203530332U (en) | 2013-11-07 | 2013-11-07 | Cell culture device capable of monitoring cell culture liquid environment |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201320697500.6U CN203530332U (en) | 2013-11-07 | 2013-11-07 | Cell culture device capable of monitoring cell culture liquid environment |
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| CN203530332U true CN203530332U (en) | 2014-04-09 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104450519A (en) * | 2014-12-09 | 2015-03-25 | 中国人民解放军兰州军区乌鲁木齐总医院 | Cell culturing system capable of precisely controlling dissolved oxygen concentration of nutrient solution |
| CN107881107A (en) * | 2017-11-15 | 2018-04-06 | 中国航天员科研训练中心 | The image capturing system of automated cell culture device |
| CN110129197A (en) * | 2018-02-08 | 2019-08-16 | 江阴瑞康健生物医学科技有限公司 | A kind of incubator with liquid circulation function |
| CN114426919A (en) * | 2020-10-29 | 2022-05-03 | 南京理工大学 | Apparatus and method for continuously monitoring cell growth rate |
-
2013
- 2013-11-07 CN CN201320697500.6U patent/CN203530332U/en not_active Expired - Lifetime
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104450519A (en) * | 2014-12-09 | 2015-03-25 | 中国人民解放军兰州军区乌鲁木齐总医院 | Cell culturing system capable of precisely controlling dissolved oxygen concentration of nutrient solution |
| CN107881107A (en) * | 2017-11-15 | 2018-04-06 | 中国航天员科研训练中心 | The image capturing system of automated cell culture device |
| CN107881107B (en) * | 2017-11-15 | 2023-06-06 | 中国航天员科研训练中心 | Image acquisition system of automatic cell culture equipment |
| CN110129197A (en) * | 2018-02-08 | 2019-08-16 | 江阴瑞康健生物医学科技有限公司 | A kind of incubator with liquid circulation function |
| CN114426919A (en) * | 2020-10-29 | 2022-05-03 | 南京理工大学 | Apparatus and method for continuously monitoring cell growth rate |
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| GR01 | Patent grant | ||
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| CX01 | Expiry of patent term |
Granted publication date: 20140409 |