CN104531618B - The method that three dimensional growth tumour cell is obtained using graphene nano material - Google Patents
The method that three dimensional growth tumour cell is obtained using graphene nano material Download PDFInfo
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- CN104531618B CN104531618B CN201410794004.1A CN201410794004A CN104531618B CN 104531618 B CN104531618 B CN 104531618B CN 201410794004 A CN201410794004 A CN 201410794004A CN 104531618 B CN104531618 B CN 104531618B
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Abstract
The invention discloses three dimensional growth tumour cell is inexpensively largely obtained using graphene nano material, the method includes at least：It will be taken out containing the culture bottle of cultured attached tumor cells, common attached cell passage operation carried out, cell suspension is made.The cell suspension inoculation of certain volume is taken respectively in new culture bottle, and the culture solution containing serum is then added in into bottle.The functional graphene material solution for having already passed through vacuum high-pressure sterilization treatment in right amount is added in into new culture bottle, after cultivating a period of time in the incubator, you can to observe the tumour cell for there are a large amount of three dimensional growths in the culture bottle for adding in functional graphene material.This method practicability is wide, raw material is taken from the grapheme material being readily synthesized, and tumor cell culture method is easy, graphene nano material solution only need to be added in than a steps more in standard passage cell incubation step, it convenient can obtain the tumour cell of three dimensional growth, cell base research and drug screening for the later stage etc., which provide, facilitates cheap pretreatment sample, greatly reduces cost and improves efficiency.
Description
Technical field
The present invention relates to medical material preparation fields, and in particular to a kind of that three-dimensional life is obtained using graphene nano material
The method of long tumour cell.The applications such as basic research and drug screening in terms of can carrying out in relation to cell.
Background technology
Using two-dimentional cell culture technology, it cannot simulate the growth of cell in vivo completely for traditional cell culture
With dynamics situation.Three-dimensional cell culture technology can observe the model of tumour formation and the effect of tumor, may be used also
The biological behaviour of observation tumour such as shifts, and can also observe interaction between cell etc., more can be in analogue body
Tumor tissues cell and microenvironment of matrix interaction caused by various situations.So Three-dimensional cell culture is a kind of completeer
The training method of U.S. analogue body inner cell situation.There are some related patents at present, such as number of patent application CN103756904A
A kind of three dimensional culture system of tumor cell culture is described, application publication number CN102952279A is described for tumour cell
The hydrogel of dimensional culture and application, it is three-dimensional thin that Authorization Notice No. CN102433258B discloses a kind of stretching-electricity combined stimulation
Born of the same parents' culture apparatus.
In the various dimensional culture technologies being currently known, reliable external magnetic field or electric field action promote cell to leave
Culture dish bottom and carry out three dimensional growth；In addition it is by being inverted culture technique to have some so that drop drops in culture dish upper table
Culture dish is overturn and cultivated by face；Also by the method for establishing three-dimensional rack so that cell is grown in passage aisle.Always
For body, although these methods preferably simulate cell three-dimensional growth pattern, the experimental skill that these methods are related to
Also in the design and making of the components such as stent, for common experimental implementation person, complicated for operation, the long-term training of needs,
It repeats, can be only achieved requirement of experiment.
The advantage of the invention is that commonly having the laboratory technician of biological cell assays technical ability, experiment can be smoothly completed, and
And it is observed that clearly three-dimensional cell grows phenomenon.Easy to operate, of low cost, experiment is convenient and efficient.
Invention content
The object of the present invention is to provide a kind of methods for the tumour cell that three dimensional growth is obtained using graphene nano material.
The present invention is achieved by the following technical solutions, includes the following steps：
1) it will be taken out containing the culture bottle of cultured attached tumor cells, carry out common attached cell passage behaviour
Make, cell suspension is made.
2) cell suspension inoculation of certain volume is taken respectively in two new culture bottles, is then added in into bottle containing serum
Culture solution.
3) toward step 2）In a new culture bottle in add in and have already passed through the graphene of vacuum high-pressure sterilization treatment in right amount
Nanomaterial solution.Another new culture bottle is added without.
4) by step 3）5% CO of two new culture bottles at 37 DEG C2A period of time is cultivated in incubator, you can with observation
There is the tumour cell of a large amount of three dimensional growths into the culture bottle for adding in graphene nano material solution；For no addition graphene
The culture bottle of nanomaterial solution, it is found that the tumour cell of adherent growth.
Further, step 2）With 3）Can exchange does not influence final result.Can graphite be added in new culture bottle in the past
Alkene nanomaterial solution adds cell suspension, is then cultivated.
Further, the attached tumor cells as described in step 1) are cultivated in culture bottle and normally behave as adherent growth
And contact inhibition.
Further, the common attached cell secondary culture operation as described in step 1), has steps of：Sop up or
It outwells with regard to culture solution；Tryptic digestive juice is added in into culture bottle to be digested；It was found that when there is gap in cell monolayer contraction
Sop up digestive juice；The culture solution containing serum is added in, cell is blown and beaten repeatedly, makes it into cell suspension.
Further, such as step 2）The cell suspension of the certain volume refers generally to step 1）In cell suspension
With 1:2 or 1:3 are dispensed.
Further, such as step 3）The graphene nano material, diameter range is 2 to 100 nanometers, while material
Functional group surface and that edge is formed containing multiple elements such as aerobic, nitrogen, sulphur, such as hydroxyl, carboxyl, carbonyl, amino, alkyl
Deng.
Further, the material of graphite content as described above includes graphite, graphite fibre, mesoporous graphite, carbon black, individual layer
Or multi-layer graphene, carbon nanotube, fullerene, also including biomaterial（Such as branch, leaf, bamboo etc.）, oil, natural gas
The material obtained Deng other flammable carbonaceous materials by the modes such as high temperature cabonization or burn incompletely collection black smoke.
Further, oxygen containing graphene nano material as described above, synthetic method are characterized in that：Stone will be contained
The carbon material of black ingredient is added in the solution of the concentrated sulfuric acid, sodium nitrate and potassium permanganate, the black suspension mixed, to it
After carrying out ultrasonic 2 hours, heat 180 degree and be stirred at reflux more than 24 hours of reaction, added in after solution is cooled to room temperature
A large amount of deionized waters adjust pH to 8 with sodium carbonate, solution are put into the bag filter that molecular cut off is 2000Da, carry out saturating
Analysis is more than 3 days, then to obtaining drying solid after the solution progress heating evaporation drying outside bag filter to get to oxygen containing stone
Black alkene nano material.As needed solid is dissolved in water again to obtain certain density graphene nano material solution.
Further, nitrogenous graphene nano material as described above, synthetic method are characterized in that：Stone will be contained
The carbon material and citric acid of black ingredient are added to nitrogenous organic solvent, such as dimethylformamide（DMF）, by black suspension
It is put into polytetrafluoroethylene (PTFE) autoclave, is heated to 200 degree and reacts more than 5 hours, after being cooled to room temperature, obtain
Filtrate is put into the bag filter that molecular cut off is 2000Da and is dialysed more than 3 by 0.22 micron of filtering membrane filtration by liquid
My god, remove extra DMF solvent, will solution carry out heating evaporation drying after obtain drying solid to get to nitrogenous graphene
Nano material.As needed solid is dissolved in water again to obtain certain density graphene nano material solution.
Further, the graphene nano material of sulfur-bearing as described above, synthetic method are characterized in that：Stone will be contained
The organic matter of the carbon material of black ingredient, citric acid and sulfur-bearing（Such as thionyl chloride, thio urea etc.）Be added to the water and ultrasound 2
A hour, obtained black suspension are added in polytetrafluoroethylene (PTFE) autoclave, are heated to 200 degree and are reacted more than 5
Hour, room temperature is then cooled to, by obtained liquid by 0.22 micron of filtering membrane filtration, second is then added in into filtrate
Alcohol, then high speed centrifugation, the drying solid that will be obtained after heat drying after obtained precipitation washing, the graphene of as sulfur-bearing are received
Rice material.As needed solid is dissolved in water again to obtain certain density graphene nano material solution.
Further, the graphene nano material of the multiple elements such as oxygen-containing, nitrogen, sulphur as described above, synthetic method
Feature may be：The synthesis step that the step of right 5,6 and 7 is subjected to arbitrary arrangement combination and is formed.For example it synthesizes
The synthesis step as described in right 6 is again carried out after graphene nano material as described in right 5.
Further, the tumour cell involved in this patent embodiment can be in Shanghai Sheng Ke institutes of Chinese Academy of Sciences cell
Resource center buys.
As described above, the present invention using graphene nano material obtain three dimensional growth tumour cell method, have with
Lower useful achievement：This method practicability is wide, the graphene nano material being readily synthesized, and tumor cell culture method is easy, only
Graphene nano material solutions need to be added in than more steps in standard passage cell incubation step, you can the convenient three dimensional growth that obtains
Tumour cell, cell base research and drug screening for the later stage etc. provides premise.All tumour cells used in the present invention
It can be commercially available and be handled without any early period.
Description of the drawings
Fig. 1 (a) and（b）The transmission electron microscope figure of graphene nano material respectively obtained in example 1,2
(TEM)。
Fig. 2 is the photoelectron spectroscopy figure (XPS) of the nitrogenous graphene nano material arrived in example 2.It can see nitrogen
Element.
Fig. 3（a）With（b）The transmission electron microscope figure (TEM) of the graphene nano material respectively obtained in example 3 and
Photoelectron spectroscopy figure (XPS).It can be seen that there is element sulphur.
Fig. 4（a）With（b）It does not add in graphene nano material in example 1 respectively and adds in graphene nano material
HeLa growth of tumour cell figures.Significantly observe（a）Figure is the tumour cell of adherent growth,（b）Figure is the tumour of three dimensional growth
Cell.
Fig. 5（a）With（b）It does not add in graphene nano material in example 2 respectively and adds in graphene nano material
A375 growth of tumour cell figures.Significantly observe（a）Figure is the tumour cell of adherent growth,（b）Figure is the tumour of three dimensional growth
Cell.
Fig. 6 is（a）With（b）It does not add in graphene nano material in example 3 respectively and adds in graphene nano material
A375 growth of tumour cell figures.Significantly observe（a）Figure is the tumour cell of adherent growth,（b）Figure is the tumour of three dimensional growth
Cell.
Fig. 7 is the ratio chart of attached cell and three dimensional growth cell under the various concentration that example 2 obtains.
Specific embodiment
Specific example is chosen according to technical solution of the present invention to be described as follows：
Embodiment 1：
By 1g graphite nanoparticles（graphite nanoparticles）It is added to the concentrated sulfuric acid, the 0.5g of 100ml 98%
In the solution of sodium nitrate and 0.2g potassium permanganate, the black suspension that is mixed, after ultrasonic 2 hours are carried out to it, heating
180 degree is simultaneously stirred at reflux more than 24 hours of reaction, adds in 800mL deionized waters after solution is cooled to room temperature, uses sodium carbonate
PH to 8 is adjusted, solution is put into the bag filter that molecular cut off is 2000Da, dialysis is carried out more than 3 days, then to bag filter
The solution of outside obtain after heating evaporation drying drying solid to get to oxygen containing graphene nano material.
It will be in 37 DEG C of 5% CO2Cultured adherent HeLa tumour cells take out in incubator, sop up or outwell as possible
Old culture solution in culture bottle, into bottle, addition trypsase is digested.When finding that cytoplasm bounces back, space between cells becomes larger, stand
Stop digestion.
Digestive juice is sucked out, a small amount of culture solution is added in bottle, residual digestive juice is rinsed out, adds the training containing serum on a small quantity
Nutrient solution.Bottle wall HeLa cells are blown and beaten repeatedly, are allowed to be detached into cell suspension.
The culture medium and about 1 × 10 of about 500 μ L is added in into 6 orifice plates5 A HeLa cells, add and have already passed through vacuum height
Press the graphene nano material solution of sterilization treatment, a concentration of 100 μ g/mL.
Culture bottle is put into 37 DEG C of 5% CO2Incubator carries out 24 hours of culture, you can glomerate to observe
The HeLa tumour cells of HeLa tumour cells, i.e. three dimensional growth.
The images of transmissive electron microscope of embodiment 1 such as Fig. 1（a）（Left figure）It is shown.It can be seen that graphene nano material is about 10
Nanometer.Cell growth figure such as Fig. 4 for not adding in and adding in graphene nano material solution of embodiment 1（a）With（b）It is shown, point
Not it is observed that apparent adherent HeLa cells and glomerate three dimensional growth tumour cell situation.
Embodiment 2：
1g carbon blacks and 0.3g citric acids are added to the dimethylformamide of 10mL（DMF）, black suspension is put into
In 25mL polytetrafluoroethylene (PTFE) autoclaves, it is heated to 200 degree and reacts more than 5 hours, after being cooled to room temperature, obtained liquid
Body is put into the bag filter that molecular cut off is 2000Da dialysis more than 3 days by 0.22 micron of filtering membrane filtration, by filtrate,
Remove extra DMF solvent, will solution carry out heating evaporation drying after obtain drying solid to get to nitrogenous graphene nano
Material.
It will be in 37 DEG C of 5% CO2Cultured adherent A375 tumour cells take out in incubator, sop up or outwell as possible
Old culture solution in culture bottle, into bottle, addition trypsase is digested.When finding that cytoplasm bounces back, space between cells becomes larger, stand
Stop digestion.
Digestive juice is sucked out, a small amount of culture solution is added in bottle, residual digestive juice is rinsed out, adds the training containing serum on a small quantity
Nutrient solution.Bottle wall A375 cells are blown and beaten repeatedly, are allowed to be detached into cell suspension.
The culture medium and about 1 × 10 of about 500 μ L is added in into 6 orifice plates5A A375 cells, add and have already passed through vacuum height
Press the graphene nano material solution of sterilization treatment, a concentration of 100 μ g/mL.
Culture bottle is put into 37 DEG C of 5% CO2Incubator carries out 24 hours of culture, you can glomerate to observe
The A375 tumour cells of A375 tumour cells, i.e. three dimensional growth.
In addition, the culture medium and about 1 × 10 of 200 μ L is added in into 6 orifice plates5 A A375 cells are added and are had already passed through very
Outage presses the graphene nano material solution of sterilization treatment, and concentration is respectively 0,25,50,75,100,150,175,200 μ g/
ML after cultivating 24 hours respectively, by adherent growth in field of view and the number of three dimensional growth tumour cell, is obtained not
With the cell proportion under growing state.
The images of transmissive electron microscope of embodiment 2 such as Fig. 1（b）（Right figure）It is shown.It can be seen that graphene nano material is about 10
Nanometer.The photoelectron spectroscopy figure of embodiment 2 is as shown in Figure 2.It can see nitrogen.Embodiment 2 does not add in and adds in graphite
Cell growth figure such as Fig. 5 of alkene nanomaterial solution（a）With（b）It is shown, respectively it is observed that apparent adherent A375 is thin
Born of the same parents and glomerate three dimensional growth tumour cell situation.Cell proportion under the different growing states of embodiment 2 is as shown in Figure 7.
Embodiment 3：
0.25g carbon blacks, 0.25g citric acids and 0.25g thio ureas are added in 20ml water and ultrasonic 2 hours, obtained
To black suspension be added in 50ml polytetrafluoroethylene (PTFE) autoclaves, be heated to 200 degree and react more than 5 hours,
Room temperature is then cooled to, by obtained liquid by 0.22 micron of filtering membrane filtration, ethyl alcohol is then added in into filtrate, then
High speed centrifugation, the graphene nano material of drying solid, as sulfur-bearing that will be obtained after heat drying after obtained precipitation washing.
It will be in 37 DEG C of 5% CO2Cultured adherent A375 tumour cells take out in incubator, sop up or outwell as possible
Old culture solution in culture bottle, into bottle, addition trypsase is digested.When finding that cytoplasm bounces back, space between cells becomes larger, stand
Stop digestion.
Digestive juice is sucked out, a small amount of culture solution is added in bottle, residual digestive juice is rinsed out, adds the training containing serum on a small quantity
Nutrient solution.Bottle wall A375 cells are blown and beaten repeatedly, are allowed to be detached into cell suspension.
The culture medium and about 1 × 10 of about 500 μ L is added in into 6 orifice plates5A A375 cells, add and have already passed through vacuum height
Press the graphene nano material solution of sterilization treatment, a concentration of 100 μ g/mL.
Culture bottle is put into 37 DEG C of 5% CO2Incubator carries out 24 hours of culture, you can glomerate to observe
The A375 tumour cells of A375 tumour cells, i.e. three dimensional growth.
The images of transmissive electron microscope of embodiment 3 such as Fig. 3（a）（Left figure）It is shown.It can be seen that graphene nano material is about 20
Nanometer.Photoelectron spectroscopy figure such as Fig. 3 of embodiment 3（b）（Right figure）It is shown.It can see element sulphur.Embodiment 3 do not add in and
Add in cell growth figure such as Fig. 6 of graphene nano material solution（a）With（b）It is shown, it is respectively it is observed that apparent adherent
A375 cells and glomerate three dimensional growth tumour cell situation.
It should be noted that herein, term " comprising ", "comprising" or its any other variant be intended to it is non-he
Property include so that process, method, article or equipment including a series of elements not only include those elements, but also
Further include other elements that are not explicitly listed or further include for this process, method, article or equipment it is intrinsic
Element.In the absence of more restrictions, the element limited by sentence "including a ...", it is not excluded that including described
Also there are other identical elements in the process of element, method, article or equipment.
The preferred embodiment of the present invention described in detail above.It should be appreciated that those of ordinary skill in the art without
Creative work is needed according to the present invention can to conceive and makes many modifications and variations.Therefore, all this technology personnel are according to this
The design of invention passes through the available technical side of logical analysis, reasoning, or a limited experiment on the basis of existing technology
Case, all should be in the protection domain being defined in the patent claims.
Claims (7)
1. the method for three dimensional growth tumour cell is obtained using graphene nano material, which is characterized in that comprise the steps of：
1) it will be taken out containing the culture bottle of cultured attached tumor cells, attached cell passage operation carried out, to be made
Cell suspension；
2) cell suspension inoculation of certain volume is taken respectively in new culture bottle, and the culture solution containing serum is then added in into bottle；
3) toward step 2）In a new culture bottle in add in and have already passed through the graphene nano of vacuum high-pressure sterilization treatment in right amount
Material solution；
4) by step 3）New culture bottle cultivate in the incubator a period of time, you can with observe add in graphene nano material
There is the tumour cell of a large amount of three dimensional growths in the culture bottle of solution；
It 5) can be with exchange step 2）With 3）And final result is not influenced, graphene nano material can be added in new culture bottle in the past
Solution adds cell suspension, is then cultivated；The graphene nano material, 2 nanometers to 100 nanometers of diameter range,
Material surface and edge is oxygen-containing, the functional group of one or more of nitrogen, element sulphur element simultaneously, the functional group is selected from hydroxyl
One or more of base, carboxyl, carbonyl, amino, alkyl.
2. obtaining the method for three dimensional growth tumour cell using graphene nano material as described in claim 1, feature exists
It is to be cultivated in culture bottle in the attached tumor cells, shows as adherent growth and contact inhibition.
3. obtaining the method for three dimensional growth tumour cell using graphene nano material as described in claim 1, feature exists
It is operated in the attached cell secondary culture, there are following steps：Sop up or outwell old culture solution；Pancreas is added in into culture bottle
Protease digestion liquid is digested；It was found that cell monolayer contraction occurs sopping up digestive juice during gap；The culture solution containing serum is added in,
Cell is blown and beaten repeatedly, makes it into cell suspension.
4. obtaining the method for three dimensional growth tumour cell using graphene nano material as described in claim 1, feature exists
In the synthetic method of the oxygen containing graphene nano material, comprise the steps of：Carbon material containing graphite content is added
Enter into the solution of the concentrated sulfuric acid, sodium nitrate and potassium permanganate, the black suspension mixed, ultrasonic 2 hours are carried out to it
Afterwards, 180 degrees Celsius are heated and is stirred at reflux more than 24 hours of reaction, solution is cooled to after room temperature and adds in a large amount of deionizations
Water adjusts pH to 8 with sodium carbonate, solution is put into the bag filter that molecular cut off is 2000Da, carries out dialysis more than 3 days,
Drying solid is obtained to get to oxygen containing graphene nano material after heating evaporation drying is then carried out to the solution outside bag filter
Material.
5. obtaining the method for three dimensional growth tumour cell using graphene nano material as described in claim 1, feature exists
In the synthetic method of the nitrogenous graphene nano material, comprise the steps of：By the carbon material containing graphite content and
Citric acid is added to nitrogenous organic solvent, and the nitrogenous organic solvent is dimethylformamide（DMF）, black is suspended
Liquid is put into polytetrafluoroethylene (PTFE) autoclave, is heated to 200 degrees Celsius and is reacted more than 5 hours, after being cooled to room temperature,
Filtrate is put into the bag filter that molecular cut off is 2000Da and is dialysed by 0.22 micron of filtering membrane filtration by obtained liquid
More than 3 days, remove extra DMF, will solution carry out heating evaporation drying after obtain drying solid to get to nitrogenous graphene
Nano material.
6. obtaining the method for three dimensional growth tumour cell using graphene nano material as described in claim 1, feature exists
In the synthetic method of the graphene nano material of the sulfur-bearing, comprise the steps of：By the carbon material containing graphite content, lemon
The organic matter of lemon acid, sulfur-bearing, the organic matter of the sulfur-bearing is one or both of thionyl chloride and thio urea, is added to water
In and ultrasonic 2 hours, obtained black suspension be added in polytetrafluoroethylene (PTFE) autoclave, be heated to 200 degrees Celsius
And more than 5 hours are reacted, and room temperature is then cooled to, it is then past by obtained liquid by 0.22 micron of filtering membrane filtration
Ethyl alcohol is added in filtrate, then high speed centrifugation, the drying solid that will be obtained after heat drying after obtained precipitation washing as contains
The graphene nano material of sulphur.
7. obtaining the method for three dimensional growth tumour cell using graphene nano material as described in claim 1, feature exists
In the synthetic method of described oxygen-containing, nitrogen, the graphene nano material of three kinds of elements of sulphur, following synthesis step is included：By right
It is required that the synthesis step that 4,5 and 6 step carries out arbitrary arrangement combination and formed.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102433258A (en) * | 2011-12-01 | 2012-05-02 | 北京航空航天大学 | Stretch-electricity combined stimulation three-dimensional cell culture device |
| CN102864119A (en) * | 2012-09-27 | 2013-01-09 | 中国科学院苏州纳米技术与纳米仿生研究所 | Carrier for cell culturing and preparation method of carrier |
| CN102952279A (en) * | 2012-05-10 | 2013-03-06 | 东南大学 | Hydrogel for three-dimensional culturing of tumor cells and applications thereof |
| CN103756904A (en) * | 2014-01-10 | 2014-04-30 | 赵海涛 | Three-dimensional culture system for tumor cell culture |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060014275A1 (en) * | 2003-09-26 | 2006-01-19 | Gsi Creos Corporation | Cell culture carrier and jig for cell culture |
| SG186313A1 (en) * | 2010-07-08 | 2013-02-28 | Univ Singapore | Method for controlling and accelerating differentiation of stem cells using graphene substrates |
-
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- 2014-12-21 CN CN201410794004.1A patent/CN104531618B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102433258A (en) * | 2011-12-01 | 2012-05-02 | 北京航空航天大学 | Stretch-electricity combined stimulation three-dimensional cell culture device |
| CN102952279A (en) * | 2012-05-10 | 2013-03-06 | 东南大学 | Hydrogel for three-dimensional culturing of tumor cells and applications thereof |
| CN102864119A (en) * | 2012-09-27 | 2013-01-09 | 中国科学院苏州纳米技术与纳米仿生研究所 | Carrier for cell culturing and preparation method of carrier |
| CN103756904A (en) * | 2014-01-10 | 2014-04-30 | 赵海涛 | Three-dimensional culture system for tumor cell culture |
Non-Patent Citations (2)
| Title |
|---|
| "Fabrication of conducting electrospun nanofibers scaffold for three-dimensional cells culture";Yashpal Sharma et al.;《International Journal of Biological Macromolecules》;20120618;第51卷;第627-631页 * |
| "Use of Three-Dimensional Tissue Cultures to Model Extravascular Transport and Predict In Vivo Activity of Hypoxia一Targeted Anticancer Drugs";Kevin O. Hicks et al.;《Journal of the National Cancer Institute》;20060816;第98卷(第16期);第1118-1128页 * |
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