CN102279186A - Method for detecting cell deformability by using poly-hydroxyethyl methacrylate (polyHEMA) - Google Patents
Method for detecting cell deformability by using poly-hydroxyethyl methacrylate (polyHEMA) Download PDFInfo
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Abstract
The invention discloses a method for detecting cell deformability by using poly-hydroxyethyl methacrylate (polyHEMA). A cell and a substrate connection part of the cell are separated partially by using the polyHEMA, so that the deformability of the cell can be estimated and detected; the deformation and growth process of the cell is recorded through a microscope; and the deformability of the cell is quantified and graded by using methods, such as a CCK8 detection method, so that the judgment on the deformability of the cell can be realized. The method has the characteristics of relative economy, simplicity, quickness, easy operation and the like.
Description
Technical field
The present invention relates to a kind of method that detects the cytomorphosis ability, relate in particular to the method that a kind of PolyHEMA of utilization detects the cytomorphosis ability, belong to the medical biotechnology field.
Background technology
The transfer of tumour is a complex processes that multistep is rapid, multifactor, sequential, and with the adhesion of tumour cell, amoeboid movement, transfer ability is closely related.Cell has experienced 7 important stages during the metastases, is respectively: 1, the tumour infinite multiplication stage; 2, intratumoral vasculature generation phase; 3, tumour cell is invaded blood vessel; 4, tumour cell drops in the blood vessel; 5, tumour is in endovascular transfer; 6, tumour cell passes blood vessel; 7, the tumour cell continued growth forms secondary tumors.Wherein, break away from original position, invade in the process of the circulation system at tumour cell; Cell is in the process that the nido attitude shifts of losing in the circulation system; At the secondary position, cell passes blood vessel, and continued growth forms in the process of secondary tumors, and tumour cell has experienced repeatedly distortion, motion, and transition process is the important behaviour mode of tumor cell invasion transfer ability to adapt to extraneous poor environment.Have only the strong tumour cell of those distortion transfer abilities to shift.In order to study the metastatic change of tumour cell, need set up easyly, differentiate the method for the deformability of tumour cell efficiently, all there is important meaning its aspect such as medicament research and development in oncology studies and control metastases.
The method of the detection tumour cell amoeboid movement ability that present stage is commonly used has: NIH3T3 cell monolayer invasion and attack empirical model (MIA); Boyden cell invasion and attack experiment; Transwell cell invasion and attack experiment and cell scratch experiment etc., the amoeboid movement of existing experimental technique pair cell, transfer ability detects all special advantages and relative weak point.
1, NIH3T3 cell monolayer invasion and attack experiments (MIA) are to detect cytomorphosis transfer ability technology commonly used now.At first, the growth of NIH3T3 cell is merged and is become cell monolayer, and cell to be measured is cultivated therewith, counts the number that each visual field penetrates the cancer cell of NIH3T3 cell, and the number of cancer cell carries out the analysis of tumour cell motion transfer ability in the unit of account area.NIH3T3 cell monolayer invasion and attack experiment still can not be got rid of influencing each other of producing between NIH3T3 cell and the detected cell.
2, Transwell cell invasion and attack experiment is that the Transwell cell is put into culture plate, deserve to be called the chamber in the cell, claim chamber down in the culture plate, the levels nutrient solution is separated by with polycarbonate membrane, with the cell kind of research last indoor, because polycarbonate membrane has permeability, composition in lower floor's nutrient solution can have influence on indoor cell, uses the polycarbonate membrane of different apertures and process different disposal, just can effectively detect cell chemotaxis, migration and invasive ability.Boyden cell invasion and attack experiment also is to detect cytomorphosis transfer ability technology commonly used.Boyden invasion and attack cell is tubbiness, and poly-carbonic acid vinegar miillpore filter is equipped with in the bottom, and the filter membrane surface scribbles artificial basilar memebrane, and invasion and attack cell and culture plate constitute the upper and lower chamber of cell invasion jointly.Cell kind to be measured is in last chamber, and following chamber then is to attract the cell chemotactic liquid of migration downwards.There is the cell of invasiveness at first to adhere to artificial basilar memebrane surface, and then the artificial basilar memebrane of secretion matrix metalloproteinase digestion degraded, and pass porous membrane by travel motion, at last attached to the filter membrane another side, the cell number that passes filter membrane can reflect the invasiveness of this cell, so Boyden cell cell culture model has the ability of comprehensive judgement tumor invasion and transfer.
Boyden cell invasion and attack experiment and Transwell cell invasion and attack experimental cost height, operation is comparatively complicated, is subject to the kinds of experiments condition effect, therefore, more than three kinds of methods in actual application, be restricted, be not suitable for to cytomorphosis campaign to be checked a large amount of Screening and Identification of transfer ability.
3, the cell scratch experiment is to operate the method that comparatively simply detects the cell movement ability.Be meant cellular incubation on double dish or flat board, treat to scrape at line of middle section picture with cell after the Fusion of Cells, the cell in this line has been got rid of by mechanical force, then cell is continued to cultivate, observation of cell is judged the transfer ability of cell to the situation of acellular scored area migration.
Though the cell scratch experiment can tentatively be judged the power of cell movement transfer ability, because in its experimentation, cell does not experience amoeboid movement, and deformability that therefore can not pair cell is made judgement.Given this, set up a kind of can be fast and effectively and the detection method that can reduce cost very necessary.
Summary of the invention
Shortcoming and defect in the authentication method of cells involved distortion transfer ability in the prior art the invention provides the method that a kind of PolyHEMA of utilization detects the cytomorphosis ability.
The method of utilizing PolyHEMA to detect the cytomorphosis ability of the present invention, step is:
(1) identifies with the PolyHEMA bag by plate with decrement method or scarification preparation; Wherein, described decrement legal system is equipped with polyHEMA bag and by the method for plate is: adding concentration in Tissue Culture Plate is the polyHEMA ethanolic solution of 12mg/ml, makes that the whole density of polyHEMA is 0.5mg/cm in the every hole of plate
2～2.5mg/cm
2, dry the Tissue Culture Plate that contains polyHEMA liquid then under the room temperature, obtain being at the bottom of the plate decrement polyHEMA plate (Figure 1A-D) in the narrow crack of network shape; Described scarification prepares polyHEMA bag: adding concentration in Tissue Culture Plate is the polyHEMA ethanolic solution of 12mg/ml, makes that the whole density of polyHEMA is 2.5mg/cm in the every hole of plate
2～8.5mg/cm
2Dry the Tissue Culture Plate that contains polyHEMA liquid then under the room temperature, use disposable syringe syringe needle or knife blade inclination 15-45 degree angle again, central cut at the polyHEMA film that has prepared, making the cut width is 10 μ m～20 μ m, obtain having at the bottom of the plate artificial cut passage cut polyHEMA plate (Fig. 2 A, B);
(2) be 1 * 10 with concentration
6Individual cell/ml～5 * 10
6In the polyHEMA plate (Fig. 3 A-F) of the cell suspension inoculation of the exponential phase of individual cell/ml to the above-mentioned sterilization, after treating that cell is even and all being paved with the polyHEMA plate, put 37 ℃, CO
2Cultivate 12～60h in the incubator; The situation of passing through and growing in the narrow crack that the microscopically observation of cell forms between polyHEMA film and Tissue Culture Plate during this time, can not freely pass through the narrow crack of polyHEMA, cell volume diminishes, and assembles agglomeratingly, and the cell that cell proliferation is blocked is judged to be the weak cell of deformability; Can pass through the narrow crack of polyHEMA, cell stretches, and becomes irregular shape, cell attachment, and the beginning proliferating cells is judged to be the strong cell of deformability; The cytomorphosis ability is strong more, and it is many more to show as the cell that passes through the narrow crack of polyHEMA, and the cell of adherent growth is many more;
(3) be 1 * 10 with concentration
6Individual cell/ml～5 * 10
6The cell suspension of the exponential phase of individual cell/ml by 100 μ L cell suspension/holes be seeded to 96 hole decrements after the sterilization or cut polyHEMA plate (Fig. 4 A, B) in, rearmounted 37 ℃ of mixing, CO
2Cultivate 12～48h in the incubator, during get 3 every group multiple holes every 12h and add 10 μ L CCK8, rearmounted 37 ℃ of mixing, CO
2Continue to cultivate 90min in the incubator; Read at the 450nm place and record data with microplate reader, draw growth curve according to data; CCK8 detects OD450nm value and is lower than OD450nm basic value 120% when rigidly connecting kind of polyHEMA plate in the growth curve, look corresponding cell be cell a little less than the deformability; CCK8 detects raise when rigidly connecting kind of polyHEMA plate OD450nm basic value 120% of OD450nm value in the growth curve, look corresponding cell be the strong cell of deformability.
In the above-mentioned steps (1), described Tissue Culture Plate is 6 porocyte culture plates and/or 96 porocyte culture plates preferably.
Above-mentioned decrement legal system is equipped with the polyHEMA bag by in the method for plate, and the whole density of polyHEMA is preferably 1.0mg/cm in every hole
2～2.0mg/cm
2
Above-mentioned scarification prepares the polyHEMA bag by in the method for plate, and the whole density of polyHEMA is preferably 3.5mg/cm in every hole
2～5.5mg/cm
2
The of the present invention utilization in the method that PolyHEMA detects the cytomorphosis ability, utilize PolyHEMA to detect the cytomorphosis ability, combine with the variation of microscopic examination cytomorphology and the detection of cytoactive and number, can effectively distinguish the power of the deformability of cell.Compare with detection method in the past, have following advantage:
1. identifying with the polyHEMA plate can be because of the evaluation of intercellular interaction influence to the tumour cell deformability, thereby can get rid of the influence of biological empirical factors such as the interaction of cell surface molecule and cell factor, the amoeboid movement ability of cell is examined in single reflection.
2. price is cheap than invasion and attack of Boyden cell and Transwell cell, and preparation identifies that the polyHEMA plate technology of usefulness is simple, easy operating, and relatively economical, easy, quick, be fit to large-scale cell drug screening experiment.
3.PolyHEMA as a kind of anionic polymer, the pair cell free of toxic effects, cell can freely pass through the formed crack of polyHEMA, and can be under the formed barrier film of polyHEMA merisis.The process that cell passes through the polyHEMA slit needs through amoeboid movement, can break away from former position by the analogue body inner tumour cell, the process of the turnover circulation system.
Description of drawings
Fig. 1 detects the preparation (decrement method) of the PolyHEMA culture plate of cytomorphosis ability
Wherein, A: microscopically is observed decrement PolyHEMA culture plate, and B: visual inspection decrement PolyHEMA culture plate, C: microscopically is observed even PolyHEMA culture plate, D: the even PolyHEMA culture plate of visual inspection.
Fig. 2 detects the preparation (scarification) of the PolyHEMA culture plate of cytomorphosis ability
Wherein, A: microscopically is observed even PolyHEMA culture plate, B: microscopically is observed cut PolyHEMA culture plate.
Fig. 3 microscopic examination cellular morphology result
Wherein, A: microscopically is observed cut PolyHEMA culture plate; B: microscopically is observed the growth (24h) of immortalization hepatoma carcinoma cell at cut PolyHEMA culture plate; C: microscopically is observed hepatoma carcinoma cell Bel7402 and is passed through cut PolyHEMA culture plate continuation propagation (24h); D: microscopically is observed hepatoma carcinoma cell Bel7402 and is passed through decrement PolyHEMA culture plate continuation propagation (24h); E: microscopically is observed the growth (24h) of immortalized hepatocyte on decrement PolyHEMA culture plate; F: microscopically is observed hepatoma carcinoma cell Bel7402 and is passed through decrement PolyHEMA culture plate continuation propagation (48h).
Fig. 4 CCK8 detects cytoactive result (growth curve)
Wherein, A: hepatoma cell line BEL7402, the growth curve of liver immortalized cells LO2 on decrement PolyHEMA culture plate, B: hepatoma cell line BEL7402, the growth curve of liver immortalized cells LO2 on cut PolyHEMA culture plate.
Embodiment
The 1PolyHEMA bag is by the preparation of plate
(1) preparation of .polyHEMA storing solution and working fluid:
(Sigma USA), is made into the solution that the polyHEMA final concentration is 120mg/ml (promptly dissolve 2.4g polyHEMA in the ethanol of 20ml95%, 65 ℃ shook mixing 8 hours, got the polyHEMA storing solution) to dissolve with ethanol polyHEMA with 95%; Then with 1: 10 by volume ratio of polyHEMA storing solution with 95% ethanol dilution, making final concentration is the polyHEMA working fluid of 12mg/ml, 4 ℃ of preservations are standby.
(2). the preparation of decrement polyHEMA plate:
The polyHEMA working fluid of getting 12mg/ml evenly is taped against 6 orifice plates (every hole floorage 9.6cm
2) the bottom surface on, the 1ml/ hole; 96 hole polyHEMA plates (every hole floorage 0.32cm
2) be 100 μ L/ holes.The whole density that makes every hole polyHEMA is 1.25mg/cm
2, dry the polyHEMA working fluid under the room temperature naturally, obtain 6 holes and 96 hole decrement polyHEMA plates (Figure 1A-D).
(3). the preparation of cut polyHEMA plate:
The polyHEMA working fluid of getting 12mg/ml evenly is taped against 6 orifice plates (every hole floorage 9.6cm
2) the bottom surface on, the 2ml/ hole; 96 hole polyHEMA plates (every hole floorage 0.32cm
2) be 200 μ L/ holes.Making the whole density of every hole polyHEMA is 2.5mg/cm
2, dry the polyHEMA working fluid under the room temperature naturally, with disposable syringe syringe needle (4.5#) or knife blade (11#) inclination 30 degree angles, central cut at the polyHEMA film that has prepared, the about 18 μ m of its width, be 6 holes and 96 hole cut polyHEMA plates (Fig. 2 A, B).
6 holes and the 96 hole polyHEMA plates of preparation decrement and cut use the preceding polyHEMA plate is placed under the uviol lamp to shine, to reach the aseptic requirement of cellular incubation.
1 cell passes through the foundation of growth model
(1). cellular incubation
1. select well-grown hepatoma carcinoma cell BEL7402 and liver immortalized cells LO2, gently the wave and culture bottle for several times, the suspension floating the fragment at cell surface, pours out together with growth-promoting media then, washes once with Hanks liquid.2. add 0.25% tryptic digestive juice, 4～5ml from acellular side, the upset culture flask makes about digestive juice submergence cell 1min.3. the culture flask that overturns is placed 5～10min, is the digestion that promotes cell, can add the digestive juice of 37 ℃ of preheatings, or cell towards on the time, pasting the bottle outer wall of cell face with palm, treat visual inspection cell face occur woven design poroid till.4. pour out digestive juice, add an amount of new prepared culture, wash cell, and with suction pipe piping and druming for several times, cell is scatter, by 1: 2 or distribution in 1: 3 cultivation of going down to posterity along the cell face.5. 37 ℃ of cultivations make cell attachment growth, form to change behind the cell monolayer to keep liquid and be for experiment again.The exponential phase cell is all adopted in all experiments.
(2). cell count and vitality test
1. the exponential phase cell is collected in digestion, and piping and druming makes cell make it to become single cell suspension as far as possible; 2. get single cell suspension, by cell: the volume ratio of trypan blue solution=9: 1 adds 0.4% trypan blue solution, and the rearmounted room temperature of mixing is about 5 minutes; 3. be ready to clean tally and cover glass, draw to dye well with the glass dropper and also blow even cell liquid gently, improved pipette tip is leaned against gently between the slit of covering glass and tally observation of cell survival rate under the mirror; 4. blood counting chamber and cover plate are tried totally with wiping, and cover plate is covered on tally.With the cell suspension sucking-off a little, drip at the cover plate edge, suspension is full of between cover plate and the tally.Left standstill 3 minutes; 5. count plate four big lattice total cellular score, line ball cell are only counted left side and top.Be calculated as follows then:
The big lattice total cellular score of cell number/ml=4/4 * 10000; Adjust cell concentration to 4 * 10
6Individual cell/ml.
(3). cell inoculation
With hepatoma carcinoma cell BEL7402, the cell suspension of liver immortalized cells LO2 is seeded to respectively in the above-mentioned polyHEMA plate (6 holes and 96 hole polyHEMA plates), and the narrow crack of the network shape of polyHEMA plate and the artificial cut passage of setting up can be decided by the size of cytomorphosis ability.Rock culture plate gently, treat even rearmounted 37 ℃ of cell shop, CO
2Cultivate 12～48h in the incubator, treat that cell begins propagation or flocculate, carries out the detection of cytomorphosis ability.
2 cells pass through the foundation of growth model
With cell inoculation in the RPMI1640 nutrient culture media that contains 10% calf serum, 37 ℃, CO
2Cultivate 24～36h in the incubator, cover with after the back is digested to single cell suspension with 37 ℃ of 0.25% pancreatin adjustment cell concentration to 4 * 10
6Individual cell/ml is seeded to (6 holes and 96 hole polyHEMA plates) in the above-mentioned polyHEMA plate, and 37 ℃, CO
2Continue to cultivate 12～48h in the incubator.
The evaluation of 3 pair cell deformabilities
(1). the detection of cell cytomorphology in crossing process
With hepatoma carcinoma cell BEL7402, liver immortalized cells LO2 is inoculated into (6 holes and 96 hole polyHEMA plates) in the above-mentioned polyHEMA plate, treats even rearmounted 37 ℃ of cell shop, CO
2Continue in the incubator to cultivate 12～48h, Taking Pictures recording is carried out in the growth of pair cell in above-mentioned polyHEMA plate.Hepatoma carcinoma cell BEL7402 in the above-mentioned polyHEMA plate of controlled observation, the process that liver immortalized cells LO2 passes through and grows in narrow crack writes down the morphological data that cell passes through with Nikon ECLIPSE Ti microscope.
(2). cell is bioactive detection in crossing process
Hepatoma carcinoma cell BEL7402 and immortalized cells LO2 are inoculated in the above-mentioned polyHEMA plate, 100 μ L/ holes, behind the mixing to 37 ℃, CO
2Continue to cultivate 48h in the incubator, get 3 every group multiple holes every 12h and add 10 μ L CCK8, behind the mixing to 37 ℃, CO
2Continue to cultivate 90min in the incubator; Read at the 450nm place and record data with the MK3 microplate reader, and draw their growth curve according to data.
(3). the judgement of identification of indicator
1. in cut polyHEMA plate (Fig. 3 A-C), liver immortalized cells LO2 can not freely pass through the narrow crack of polyHEMA, thereby cell aggregation is agglomerating, smaller volume, cell proliferation is subjected to retardance (Fig. 3 B); And hepatoma carcinoma cell BEL7402 has stronger deformability, can effectively pass through the narrow crack of polyHEMA, and continues adherent growth and pass through the narrow crack of polyHEMA, and cell stretches, and becomes irregular shape, and cell begins propagation; (Fig. 3 C).In decrement polyHEMA plate (Fig. 3 D-F), liver immortalized cells LO2 can not freely pass through the narrow crack of polyHEMA, thereby cellular morphology presents gathering agglomerating (Fig. 3 E), and hepatoma carcinoma cell BEL7402 has stronger deformability, can effectively pass through the narrow crack that polyHEMA forms, and the continuation adherent growth (Fig. 3 D, F).
To sum up the result in decrement polyHEMA plate (Fig. 4 A) and cut polyHEMA plate (Fig. 4 B), can judge that by the form of cell the deformability of hepatoma carcinoma cell BEL7402 is strong, and a little less than the deformability of immortalized cells LO2.
2. the growth curve of cell shows, in decrement polyHEMA plate (Fig. 4 A) and cut polyHEMA plate (Fig. 4 B), hepatoma carcinoma cell BEL7402 continues adherent growth, and the OD450nm value during CCK8 detects continues conspicuousness and raises, when rigidly connecting kind of polyHEMA plate 120% of the OD450nm basic value; And immortalized cells LO2 does not see cell proliferation, and the OD450nm value rising during CCK8 detects is lower than 120% of OD450nm basic value when rigidly connecting kind of polyHEMA plate.
Growth curve by above-mentioned cell can be judged: the cytomorphosis ability of hepatoma carcinoma cell BEL7402 is better than immortalized cells LO2.
Claims (4)
1. method of utilizing PolyHEMA to detect the cytomorphosis ability, step is:
(1) identifies with the PolyHEMA bag by plate with decrement method or scarification preparation; Wherein, described decrement legal system is equipped with polyHEMA bag and by the method for plate is: adding concentration in Tissue Culture Plate is the polyHEMA ethanolic solution of 12mg/ml, makes that the whole density of polyHEMA is 0.5mg/cm in the every hole of plate
2～2.5mg/cm
2, dry the Tissue Culture Plate that contains polyHEMA liquid then under the room temperature, obtain being at the bottom of the plate decrement polyHEMA plate in the narrow crack of network shape; Described scarification prepares polyHEMA bag: adding concentration in Tissue Culture Plate is the polyHEMA ethanolic solution of 12mg/ml, makes that the whole density of polyHEMA is 2.5mg/cm in the every hole of plate
2～8.5mg/cm
2Dry the Tissue Culture Plate that contains polyHEMA liquid then under the room temperature, use disposable syringe syringe needle or knife blade inclination 15-45 degree angle again, central cut at the polyHEMA film that has prepared, making the cut width is 10 μ m～20 μ m, obtains having at the bottom of the plate cut polyHEMA plate of artificial cut passage;
(2) be 1 * 10 with concentration
6Individual cell/ml～5 * 10
6In the polyHEMA plate of the cell suspension inoculation of the exponential phase of individual cell/ml to the above-mentioned sterilization, after treating that cell is even and all being paved with the polyHEMA plate, put 37 ℃, CO
2Cultivate 12～60h in the incubator; The situation of passing through and growing in the narrow crack that the microscopically observation of cell forms between polyHEMA film and Tissue Culture Plate during this time, can not freely pass through the narrow crack of polyHEMA, cell volume diminishes, and assembles agglomeratingly, and the cell that cell proliferation is blocked is judged to be the weak cell of deformability; Can pass through the narrow crack of polyHEMA, cell stretches, and becomes irregular shape, cell attachment, and the beginning proliferating cells is judged to be the strong cell of deformability; The cytomorphosis ability is strong more, and it is many more to show as the cell that passes through the narrow crack of polyHEMA, and the cell of adherent growth is many more;
(3) be 1 * 10 with concentration
6Individual cell/ml～5 * 10
6The cell suspension of the exponential phase of individual cell/ml is seeded in the 96 hole decrements or cut polyHEMA plate after the sterilization by 100 μ L cell suspension/holes, rearmounted 37 ℃ of mixing, CO
2Cultivate 12～48h in the incubator, during get 3 every group multiple holes every 12h and add 10 μ L CCK8, rearmounted 37 ℃ of mixing, CO
2Continue to cultivate 90min in the incubator; Read at the 450nm place and record data with microplate reader, draw growth curve according to data; CCK8 detects OD450nm value and is lower than OD450nm basic value 120% when rigidly connecting kind of polyHEMA plate in the growth curve, be considered as corresponding cell be cell a little less than the deformability; CCK8 detects raise when rigidly connecting kind of polyHEMA plate OD450nm basic value 120% of OD450nm value in the growth curve, be considered as corresponding cell be the strong cell of deformability.
2. utilize PolyHEMA to detect the method for cytomorphosis ability according to claim 1, it is characterized in that: described Tissue Culture Plate is 6 porocyte culture plates and/or 96 porocyte culture plates.
3. utilize PolyHEMA to detect the method for cytomorphosis ability according to claim 1, it is characterized in that: described decrement legal system is equipped with the polyHEMA bag by in the method for plate, and the whole density of polyHEMA is 1.0mg/cm in every hole
2～2.0mg/cm
2
4. utilize PolyHEMA to detect the method for cytomorphosis ability according to claim 1, it is characterized in that: described scarification prepares the polyHEMA bag by in the method for plate, and the whole density of polyHEMA is 3.5mg/cm in every hole
2～5.5mg/cm
2
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| CN1490400A (en) * | 2002-10-17 | 2004-04-21 | 中国科学院力学研究所 | Micro-carpillary tube covering method for controlling cell special distribution in shape and size, and use thereof |
| CN101182492A (en) * | 2007-11-02 | 2008-05-21 | 山东大学 | Vitro model for dynamic modeling human glioma cell displace |
| CN101182491A (en) * | 2007-11-02 | 2008-05-21 | 山东大学 | Vitro model capable of Dynamic modeling human cancer of liver displace |
| CN101205529A (en) * | 2007-11-02 | 2008-06-25 | 山东大学 | Method for constructing external model dynamically simulating human liver cancer metastasis |
| CN101724604A (en) * | 2009-12-16 | 2010-06-09 | 山东大学 | Method for acquiring solid tumor cell with successful in-vitro transfection |
| WO2011034413A1 (en) * | 2009-09-18 | 2011-03-24 | Mimos Berhad | A nitrate sensor |
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Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1490400A (en) * | 2002-10-17 | 2004-04-21 | 中国科学院力学研究所 | Micro-carpillary tube covering method for controlling cell special distribution in shape and size, and use thereof |
| CN101182492A (en) * | 2007-11-02 | 2008-05-21 | 山东大学 | Vitro model for dynamic modeling human glioma cell displace |
| CN101182491A (en) * | 2007-11-02 | 2008-05-21 | 山东大学 | Vitro model capable of Dynamic modeling human cancer of liver displace |
| CN101205529A (en) * | 2007-11-02 | 2008-06-25 | 山东大学 | Method for constructing external model dynamically simulating human liver cancer metastasis |
| WO2011034413A1 (en) * | 2009-09-18 | 2011-03-24 | Mimos Berhad | A nitrate sensor |
| CN101724604A (en) * | 2009-12-16 | 2010-06-09 | 山东大学 | Method for acquiring solid tumor cell with successful in-vitro transfection |
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