CN201746554U - Novel electrical field bioreactor used for wound healing research - Google Patents
Novel electrical field bioreactor used for wound healing research Download PDFInfo
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- CN201746554U CN201746554U CN2010201850759U CN201020185075U CN201746554U CN 201746554 U CN201746554 U CN 201746554U CN 2010201850759 U CN2010201850759 U CN 2010201850759U CN 201020185075 U CN201020185075 U CN 201020185075U CN 201746554 U CN201746554 U CN 201746554U
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Abstract
The utility model discloses a novel electrical field bioreactor used for wound healing research, and relates to a bioreactor device. The novel bioreactor device comprises a cell culture room, two cell culture medium pools, two agar salt bridges and two Steinberger solution pools, wherein the two cell culture medium pools are positioned at the two ends of the cell culture room sealed by a culture room cover; a gap is formed between the culture room cover and the bottom of the cell culture room and forms a cuboid cavity; the cavity is communicated with the cell culture medium pools at the two ends; two electric field testing probes are respectively inserted into the two cell culture medium pools; the two cell culture medium pools are connected with the Steinberger solution pools through the agar salt bridges respectively; two carbon electrodes are respectively inserted into the two Steinberger solution pools; and the carbon electrodes are in wire connection with an electric field source. The electric field is steady and uniform, and can be applied to culture cells for a long period in a nontoxic and harmless manner; and the electrical field bioreactor device has the advantages of real-time observation and is simple and convenient for operation.
Description
Technical field
The utility model relates to a kind of bioreactor device, relates in particular to a kind of novel electric field bioreactor that is used for researching wound healing.
Background technology
After epithelium is injured, can produce an endogenous electric field, this electric field pair cell healing plays a part very important.Study the reaction of cell, thereby further understand the mechanism of cell healing biological DC electric field.But in actually operating, the investigator is difficult to directly with wound as research object, so can carry out cytology research by electric field in the simulated tissue or organ culture.The experimental installation of existing a kind of galvanotaxis culturing room is such: the tissue culture ware of standard, and the inside is full of silicone grease, two long strip shape glass partition 1cm, is fixed on the culture dish bottom in parallel to each other; The groove that cell places glass cover noted earlier to form also has a glass cover to be fixed on the silicone grease top and goes up across whole groove; The aqueous paper slip of bag is placed in the plate to keep humidity; Two 10cm are long, the insides are full of the brinish Glass tubing that contains 2% agar, i.e. salt bridge, and the hole of passing respectively on the glass cover is electrically connected with culturing room; The Ag/AgCl electrode all is connected with direct supply in the beaker.One end of Glass tubing places the nutrient solution of cell, and the other end places the salt solution of beaker.By this device can observation of cell to the reaction of electric field.The galvanotaxis culturing room of another kind of using electric field: the large container control that the electric field experiment is made up of two resin glass culture dish, two culture dish connect by a throat, and platinum wire electrode places the opposite end of container respectively.Be full of the gel of the cell culture fluid composition of dissolving 2% agar in the container, the connecting passage top that is full of nutrient solution is fixed and be installed in to the glass cover and the agar that have attached cell.Glass cover is provided with the glass vision panel that can observe connecting passage.But there is following shortcoming in these devices:
One, the electric field at cell culture chamber two ends is not a parallel electric field, be the equal of an electric field, salt bridge connects cell culture medium and power supply, and an end of salt bridge tube forms point source, therefore the middle portion of culture dish only being arranged is parallel electric field, and other parts are not the electric field of a homogeneous.When two electrodes directly inserted nutrient agar, the electric field in the cell culture chamber neither the homogeneous electric field.In addition, electrode directly inserts nutrient agar, and joule heating concentrates on the two poles of the earth, can not be distributed in real time, and the time has been grown the connection that will change between electrode and the nutrient agar, causes the electric field instability in the cell culture medium, and direction of an electric field is inconsistent;
Two, thereby all culturing room's glass cover is fixed by silicone grease and is formed a fixedly geometric space, and silicone grease directly contacts with cell, and this will influence the cytoactive of substratum the inside;
Three, salt bridge is enclosed in the Glass tubing, and occupation space is very big, is unfavorable for real-time cell observation;
Four, long electrical field stimulation can change the pH value, temperature and the chemical densities that connect the liquid level between cell culture chamber and the salt bridge;
Five, Ag/AgCl electrode product pair cell is deleterious, can enter in the cell culture chamber by salt bridge in test, influences cytoactive.
The utility model content
At the problem of above-mentioned prior art existence, but the utility model purpose is to provide a kind of electric field stable uniform, nontoxic ground long duration of action in culturing cell, and can electric field bioreactor device real-time monitored, simple to operation.
The technical solution of the utility model is: a kind of novel electric field bioreactor that is used for researching wound healing, comprise a cell culture chamber, two cell culture medium ponds, two agar bridges and two Staenberg solution (steinberg solution, literal translation is Staenberg solution) pond, described two cell culture medium ponds are positioned at by the cell culture chamber two ends of cultivating the chamber cap sealing, interbody spacer one gap forms a rectangular parallelepiped cavity at the bottom of cultivating chamber cap and cell culture chamber, this cavity is communicated with the cell culture medium pond at two ends, be inserted with electrical field test probe in the two cell culture medium ponds respectively, described two cell culture medium ponds are connected with a Staenberg solution pool by an agar bridge respectively, the two ends of described agar bridge are immersed respectively in cell culture medium pond and the Staenberg solution pool, and the agar bridge two ends are the rectangular plate that parallels, the also parallel symmetry of agar bridge termination rectangular plate at cell culture chamber two ends, also be inserted with a carbon dioxide process carbon electrode in the two Staenberg solution pools respectively, the carbon dioxide process carbon electrode lead connects electric field source.
As preferably, comprise base plate and cover plate that a resin glass is made, the cover plate fixed interval is in the base plate top, base plate is provided with five grooves and constitutes a cell culture chamber, two cell culture medium ponds and two Staenberg solution pools, and the groove that is communicated with cell culture medium pond and Staenberg solution pool is full of the salt agar gel and constitutes described agar bridge.
Further, cultivate interbody spacer 300 μ m at the bottom of chamber cap and the septal cell culturing room described in above-mentioned each scheme.
The beneficial effects of the utility model are: by the observation of cell cultures, culturing cell migration etc., confirm that this bioreactor device can repeat the research of foreign literature report, the electric field stable uniform, easy and simple to handle, stable performance can be applicable to cells involved to the biomedicine of electric field reaction and the research of biophysics aspect.
Description of drawings
Fig. 1 is the sectional view of the utility model embodiment 1;
Fig. 2 is the vertical view of Fig. 1;
Fig. 3 is the vertical view of the utility model embodiment 2;
The track that Fig. 4 moves under electric field action for cell (the diagram left side is negative electrode);
Fig. 5 is the statistics (X, the parasang of Y-axis are μ m, and the diagram left side is a negative electrode) of cell traveling locus under electric field action.
Embodiment
As a kind of embodiment of the present utility model, a kind of novel electric field bioreactor that is used for researching wound healing, comprise a cell culture chamber 4, two cell culture medium ponds 3, two agar bridges 6 and two Staenberg solution pools 2, described two cell culture medium ponds 3 are positioned at by cell culture chamber 4 two ends of cultivating chamber cap 5 sealings, cultivate chamber cap 5 and interbody spacer one gap, 4 end of cell culture chamber and form a rectangular parallelepiped cavity, this cavity is communicated with the cell culture medium pond 3 at two ends, be inserted with electrical field test probe 10 in the two cell culture medium ponds 3 respectively, described two cell culture medium ponds 3 are connected with a Staenberg solution pool 2 by an agar bridge 6 respectively, the two ends of described agar bridge 6 are immersed respectively in cell culture medium pond 3 and the Staenberg solution pool 2, agar bridge 6 forbids that any exchange is arranged between Liang Chi, and agar bridge 6 two ends are the rectangular plate that parallels, the also parallel symmetry of agar bridge 6 termination rectangular plates at cell culture chamber 4 two ends, just as a battery lead plate, so cell culture chamber 4 is exposed in the electric field.Agar bridge 6 terminations cut into rectangular plate to obtain parallel EF, make sense of current unanimity between agar bridge 6 two ends and the cell culture chamber 4, and the electric field in the cell culture chamber 4 is unified parallel electric field.Also be inserted with a carbon dioxide process carbon electrode 8 in the two Staenberg solution pools 2 respectively, carbon dioxide process carbon electrode 8 leads connect electric field source 9.Electrode byproduct in the Staenberg solution pool 2 is a carbon, and is harmless to substratum.Described cultivation chamber cap 5 and 4 end of cell culture chamber interbody spacer 300 μ m.
As preferably, embodiment 1 as depicted in figs. 1 and 2, comprise base plate 1 and cover plate 7 that a resin glass is made, cover plate 7 fixed interval are in base plate 1 top, base plate 1 is provided with five grooves and constitutes a cell culture chamber 4, two cell culture medium ponds 3 and two Staenberg solution pools 2, and the groove that is communicated with cell culture medium pond 3 and Staenberg solution pool 2 is full of the salt agar gel and constitutes described agar bridge 6.
Another embodiment 2, as shown in Figure 3, for economy system shared space in the cell thermostat container, agar bridge 6 can also be bent into the L type, thereby reduce the length of base plate 1, in order to keep normal carbonic acid gas and to discharge water electrolytic gas, cover plate 7 is fixed on from the place of base plate 1 high 3mm by 8 plastic screws, and all the other are identical with embodiment 1.
In order to confirm the function of this novel galvanotaxis bio-reactor, human body epithelial cell sequence Hy926 was cultivated 16 hours, then in online timing microscope (Olympus CCD microscopic system) observation of cell migration down.Write down the cell migration track and it is handled by Image-pro plus5.0 (IPP5.0) analysis software.There is the cell directional migration in the experimental result demonstration, and cell has the galvanotaxis behavior (see figure 4) to negative electrode.Come the computation migration distance by IPP5.0, by Microsoft EXCEL 2000 gained cell migration data are converted to scatter diagram (as shown in Figure 5) then, the left side of Fig. 4 and Fig. 5 is a negative electrode, and obviously, most cells is to cathodic migration.
Claims (3)
1. novel electric field bioreactor that is used for researching wound healing, it is characterized in that: comprise a cell culture chamber, two cell culture medium ponds, two agar bridges and two Staenberg solution pools, described two cell culture medium ponds are positioned at by the cell culture chamber two ends of cultivating the chamber cap sealing, interbody spacer one gap forms a rectangular parallelepiped cavity at the bottom of cultivating chamber cap and cell culture chamber, this cavity is communicated with the cell culture medium pond at two ends, be inserted with electrical field test probe in the two cell culture medium ponds respectively, described two cell culture medium ponds are connected with a Staenberg solution pool by an agar bridge respectively, the two ends of described agar bridge are immersed respectively in cell culture medium pond and the Staenberg solution pool, and the agar bridge two ends are the rectangular plate that parallels, the also parallel symmetry of agar bridge termination rectangular plate at cell culture chamber two ends, also be inserted with a carbon dioxide process carbon electrode in the two Staenberg solution pools respectively, the carbon dioxide process carbon electrode lead connects electric field source.
2. a kind of novel electric field bioreactor that is used for researching wound healing according to claim 1, it is characterized in that: comprise base plate and cover plate that a resin glass is made, the cover plate fixed interval is in the base plate top, base plate is provided with five grooves and constitutes a cell culture chamber, two cell culture medium ponds and two Staenberg solution pools, and the groove that is communicated with cell culture medium pond and Staenberg solution pool is full of the salt agar gel and constitutes described agar bridge.
3. a kind of novel electric field bioreactor that is used for researching wound healing according to claim 1 and 2 is characterized in that: interbody spacer 300 μ m at the bottom of described cultivation chamber cap and the septal cell culturing room.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010201850759U CN201746554U (en) | 2010-05-10 | 2010-05-10 | Novel electrical field bioreactor used for wound healing research |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010201850759U CN201746554U (en) | 2010-05-10 | 2010-05-10 | Novel electrical field bioreactor used for wound healing research |
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| Publication Number | Publication Date |
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| CN201746554U true CN201746554U (en) | 2011-02-16 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN2010201850759U Expired - Fee Related CN201746554U (en) | 2010-05-10 | 2010-05-10 | Novel electrical field bioreactor used for wound healing research |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103525702A (en) * | 2013-10-30 | 2014-01-22 | 中国人民解放军第二军医大学 | Observation device for cell biological behaviors in direct current electric field |
-
2010
- 2010-05-10 CN CN2010201850759U patent/CN201746554U/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103525702A (en) * | 2013-10-30 | 2014-01-22 | 中国人民解放军第二军医大学 | Observation device for cell biological behaviors in direct current electric field |
| CN103525702B (en) * | 2013-10-30 | 2014-12-10 | 中国人民解放军第二军医大学 | Observation device for cell biological behaviors in direct current electric field |
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| Date | Code | Title | Description |
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| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20110216 Termination date: 20130510 |