CN112715435A - Method for preparing zebra fish thrombus model by using ferric chloride - Google Patents
Method for preparing zebra fish thrombus model by using ferric chloride Download PDFInfo
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- 241000252212 Danio rerio Species 0.000 title claims abstract description 63
- 208000007536 Thrombosis Diseases 0.000 title claims abstract description 50
- 229910021578 Iron(III) chloride Inorganic materials 0.000 title claims abstract description 17
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 title claims abstract description 15
- 238000000034 method Methods 0.000 title claims abstract description 12
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims abstract description 34
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 23
- JRBJSXQPQWSCCF-UHFFFAOYSA-N 3,3'-Dimethoxybenzidine Chemical compound C1=C(N)C(OC)=CC(C=2C=C(OC)C(N)=CC=2)=C1 JRBJSXQPQWSCCF-UHFFFAOYSA-N 0.000 claims abstract description 19
- 238000010186 staining Methods 0.000 claims abstract description 18
- 238000004043 dyeing Methods 0.000 claims abstract description 15
- 239000000243 solution Substances 0.000 claims abstract description 15
- 241001098657 Pterois Species 0.000 claims abstract description 13
- 238000012258 culturing Methods 0.000 claims abstract description 13
- 239000012192 staining solution Substances 0.000 claims abstract description 13
- 210000001161 mammalian embryo Anatomy 0.000 claims abstract description 9
- 239000011521 glass Substances 0.000 claims abstract description 7
- 238000005406 washing Methods 0.000 claims abstract description 7
- 230000000694 effects Effects 0.000 claims abstract description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 14
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 12
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 8
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 6
- 239000001632 sodium acetate Substances 0.000 claims description 6
- 239000011780 sodium chloride Substances 0.000 claims description 6
- 238000010171 animal model Methods 0.000 claims description 5
- 229940079593 drug Drugs 0.000 claims description 5
- 239000003814 drug Substances 0.000 claims description 5
- 235000017281 sodium acetate Nutrition 0.000 claims description 4
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 2
- 238000000465 moulding Methods 0.000 abstract description 2
- 210000002257 embryonic structure Anatomy 0.000 description 7
- 238000005516 engineering process Methods 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- HKOOXMFOFWEVGF-UHFFFAOYSA-N phenylhydrazine Chemical compound NNC1=CC=CC=C1 HKOOXMFOFWEVGF-UHFFFAOYSA-N 0.000 description 5
- 229940067157 phenylhydrazine Drugs 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 3
- 241000251468 Actinopterygii Species 0.000 description 2
- 206010003178 Arterial thrombosis Diseases 0.000 description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 206010047249 Venous thrombosis Diseases 0.000 description 2
- 210000001367 artery Anatomy 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
- 229910001628 calcium chloride Inorganic materials 0.000 description 2
- 238000012136 culture method Methods 0.000 description 2
- FBAFATDZDUQKNH-UHFFFAOYSA-M iron chloride Chemical compound [Cl-].[Fe] FBAFATDZDUQKNH-UHFFFAOYSA-M 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- VTLYFUHAOXGGBS-UHFFFAOYSA-N Fe3+ Chemical compound [Fe+3] VTLYFUHAOXGGBS-UHFFFAOYSA-N 0.000 description 1
- 208000010378 Pulmonary Embolism Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 210000001772 blood platelet Anatomy 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 229910001447 ferric ion Inorganic materials 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- NMCUIPGRVMDVDB-UHFFFAOYSA-L iron dichloride Chemical group Cl[Fe]Cl NMCUIPGRVMDVDB-UHFFFAOYSA-L 0.000 description 1
- 208000028867 ischemia Diseases 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000012452 mother liquor Substances 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000004088 pulmonary circulation Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 208000019553 vascular disease Diseases 0.000 description 1
- 210000003556 vascular endothelial cell Anatomy 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
- A61K49/0008—Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/40—Fish
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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- Zoology (AREA)
- Endocrinology (AREA)
- Rheumatology (AREA)
- Animal Husbandry (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Diabetes (AREA)
- Marine Sciences & Fisheries (AREA)
- Gastroenterology & Hepatology (AREA)
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Abstract
The invention relates to a method for preparing a zebra fish thrombus model by using ferric chloride, which comprises the following steps: 1) selecting about 30 normal-developing 2dpf zebra fishes, transferring into 0.1-1.1% ferric chloride solution prepared by embryo culture water, culturing in an incubator for 48h, removing culture water, and dyeing in 6 micro-porous plates with o-dianisidine dyeing solution in a dark place for 15 min; 2) removing the staining solution, and rapidly washing with DMSO for 3 times; 3) transferring the zebra fish into a new 6-micropore plate, and adding equal volume of DMSO (dimethyl sulfoxide); 4) placing the zebra fish on a glass slide, and taking a picture of the zebra fish under a microscope and storing the zebra fish; 5) calculating the body part and thrombus staining area of the zebra fish by using Image-ProPlus6, wherein (thrombus staining area/body area) × 100% is used as an index of thrombus occurrence degree; the invention has the advantages of cheap reagent, stable molding rate and good film forming effect.
Description
Technical Field
The invention belongs to the field of medicines, and particularly relates to a method for preparing a zebra fish thrombus model by using ferric chloride.
Background
Thrombosis is caused by multiple factors of heredity and environment and can occur in arteries and veins, the venous thrombosis is red thrombosis which can cause limb swelling and pain, the venous thrombosis falls off and can enter pulmonary circulation to cause pulmonary artery embolism, serious patients can have difficulty in breathing and even die, the arterial thrombosis is white thrombosis, ischemia change of limbs and important visceral organs can be caused, the arterial thrombosis falls off and can block the distal artery along with blood flow, limb or organ necrosis is caused, the harm of the thrombosis to human bodies is large, active treatment is needed, at present, because of the reasons of large social pressure, serious environmental pollution, bad life style and the like, the trend of thrombosis youthful development is increasingly intensified.
The zebra fish embryo is transparent and develops in vitro, which is a good model for researching vascular diseases. At present, the research on the zebra fish thrombus model is less, the number of selectable methods is small, and the invention aims to provide a novel preparation method of the zebra fish thrombus model.
Disclosure of Invention
After ferric chloride treatment, many aggregates formed by ferric ions are collected on the vascular endothelial cells, and the aggregates are attached to blood platelets to form thrombus. The preparation process of the invention is as follows:
1. selecting about 30 normal-developing 2dpf zebra fishes, transferring into 0.1-1.1% ferric chloride solution prepared from embryo culture water, culturing in an incubator for 48h, removing culture water, and dyeing in 6 micro-porous plates with o-dianisidine dyeing solution in a dark place for 15 min;
2. removing the staining solution, and rapidly washing with DMSO for 3 times;
3. transferring the zebra fish into a new 6-micropore plate, and adding equal volume of DMSO (dimethyl sulfoxide);
4. placing the zebra fish on a glass slide, and taking a picture of the zebra fish under a microscope and storing the zebra fish;
5. calculating the body part and thrombus staining area of the zebra fish by using Image-Pro Plus6, wherein the value of (thrombus staining area/body area) × 100% is used as an index of thrombus occurrence degree;
the formula of the water for culturing the zebra fish comprises the following components: 5mmol/L NaCl, 0.17mmol/L KCl, 0.4mmol/L CaCl2，0.16mmol/L MgSO4；
The formula of the o-dianisidine staining solution comprises the following components: contains 0.6mg/mL o-dianisidine, 0.01mol/L sodium acetate, and 0.65% H2O2，0.16mmol/LMgSO4。
Preferably, the method for preparing the animal model of the invention comprises the following steps:
1. selecting about 30 normal-developing 2dpf zebra fishes, transferring into 0.7% ferric chloride solution prepared by embryo culture water, culturing in an incubator for 48h, removing the culture water, and dyeing in 6 micro-porous plates for 15min in a dark place by using an o-dianisidine dyeing solution;
2. removing the staining solution, and rapidly washing with DMSO for 3 times;
3. transferring the zebra fish into a new 6-micropore plate, and adding equal volume of DMSO (dimethyl sulfoxide);
4. placing the zebra fish on a glass slide, and taking a picture of the zebra fish under a microscope and storing the zebra fish;
5. calculating the body part and thrombus staining area of the zebra fish by using Image-Pro Plus6, wherein the value of (thrombus staining area/body area) × 100% is used as an index of thrombus occurrence degree;
the formula of the water for culturing the zebra fish comprises the following components: 5mmol/L NaCl, 0.17mmol/L KCl, 0.4mmol/L CaCl2，0.16mmol/L MgSO4；
The formula of the o-dianisidine staining solution comprises the following components: contains 0.6mg/mL o-dianisidine, 0.01mol/L sodium acetate, and 0.65% H2O2，0.16mmol/LMgSO4。
Another object of the present invention is to provide a method for using the zebrafish thrombosis animal model:
1. preparing a zebra fish thrombus animal model according to the above contents;
2. adding the drug to be tested, and continuing culturing for a corresponding time;
3. dyeing for 15min by using an o-dianisidine dyeing solution;
4. the zebrafish trunk and thrombus staining area (thrombus staining area/trunk area) × 100% value was calculated using Image-Pro Plus6 as an index for evaluating the effect of the drug on thrombus.
The invention has the beneficial effects that:
the method for preparing the zebra fish thrombus model by using the ferric chloride is obtained, and has the advantages of cheap reagent, stable molding rate and good film forming effect.
Drawings
FIG. 1: tail thrombus, A, control group B, 0.7% FeCl3Solution set
FIG. 2: ratio of area of thrombus at tail to area of trunk
FIG. 3: mortality of zebra fish
Detailed Description
The present invention is further illustrated in detail by the following examples, which are provided only for illustrating the present invention and are not intended to limit the scope of the present invention.
Example 1: preparation of animal models of the invention
Experimental materials: DMSO, o-dianisidine from SIGMA; FeCl3、NaCl、KCl、CaCl2、MgSO4Sodium acetate, H2O2From Shanghai Ji to Biochemical technology, Inc.; the 6-well plate was purchased from Beijing Lvbao scientific development Co., Ltd.
An experimental instrument: the Leica DVM6 microscope is available from Beijing Rake Zhongji science and technology Co., Ltd; constant temperature incubator JF-JGDW20-100 was purchased from Shanghai Jingfu instruments, Inc.
Zebra fish breeding environment: zebra fish for experiment is AB system, and is bred by Westefield culture method in Chiba Suffolk science and technology (Guangzhou) limited with water temperature of 28 ℃, conductivity of 400-.
The experimental method comprises the following steps: selecting healthy and mature AB-series zebra fishes, putting the AB-series zebra fishes into a fish tank according to the proportion of male and female 2:1, obtaining embryos in the morning of the next day, selecting healthy and high-quality embryos, putting the healthy and high-quality embryos into an incubator at 28 ℃, changing water twice a day, selecting 30 zebra fishes which normally develop respectively in a blank control group and a model control group when the zebra fishes develop to 2dpf, wherein the blank control group is embryo culture water, and the model control group is added with FeCl which is prepared from the embryo culture water by 0.1%, 0.3%, 0.5%, 0.7%, 0.9% and 1.1% in mass fraction3Continuously culturing the solution for 48h, sucking culture water, sucking isovolume o-dianisidine staining solution by using a pipette gun, and dyeing in a 6-micropore plate for 15min in a dark place; removing the staining solution, and rapidly washing with DMSO for 3 times; putting zebra fish into a new 6-micropore plate, and adding equal volume of DMSO (dimethyl sulfoxide); taking the zebra fish, placing the zebra fish on a glass slide, and taking a picture of the zebra fish under a microscope and storing the zebra fish; the body parts and thrombus staining areas of zebra fish (thrombus staining area/body area) × 100% values were calculated using Image-Pro Plus6 as indicators of the degree of thrombus formation, and the results are shown in FIGS. 1 and 2, and the death rate of embryos of each group at the end of the experiment is shown in FIG. 3.
As can be seen from FIGS. 1 and 2, FeCl3The solution can cause the formation of thrombus in zebrafish, 0.7% and 0.9% and 1.1% (thrombus staining area/body)Dry area) × 100% are not very different, but the mortality rate of zebrafish is greatly increased, so 0.7% is the optimum dose.
Example 2 comparison of this model with the phenylhydrazine induction model
Experimental materials: DMSO, o-dianisidine from SIGMA; FeCl3、NaCl、KCl、CaCl2、MgSO4Sodium acetate, H2O2From Shanghai Ji to Biochemical technology, Inc.; the 6-hole plate is purchased from Beijing Lvurucao scientific development Co., Ltd; phenylhydrazine was purchased from Kyoto pure technologies, Inc. and prepared into 1.5mmol/L mother liquor with 100% ethanol.
An experimental instrument: the Leica DVM6 microscope is available from Beijing Rake Zhongji science and technology Co., Ltd; constant temperature incubator JF-JGDW20-100 was purchased from Shanghai Jingfu instruments, Inc.
Zebra fish breeding environment: zebra fish for experiment is AB system, and is bred by Westefield culture method in Chiba Suffolk science and technology (Guangzhou) limited with water temperature of 28 ℃, conductivity of 400-.
The experimental method comprises the following steps: selecting healthy mature AB-series zebra fishes, putting the healthy mature AB-series zebra fishes into a fish tank according to the proportion of male and female parts being 2:1, obtaining embryos in the morning of the next day, selecting healthy high-quality embryos, putting the healthy high-quality embryos into an incubator at 28 ℃, changing water twice a day, selecting 30 normally-developed zebra fishes from a blank control group, an iron chloride model group and a phenylhydrazine model group when the zebra fishes develop to 2dpf, wherein the blank control group is embryo culture water, the mass fraction of the iron chloride model group is 0.7%, the concentration of the phenylhydrazine model group is 1.5 mu mol/L, continuously culturing for 48h, sucking the culture water, sucking an equal volume of o-dianisidine staining solution by using a liquid transfer gun, and staining for 15min in a 6 micropore plate in a dark place; removing the staining solution, and rapidly washing with DMSO for 3 times; transferring the zebra fish into a new 6-micropore plate, and adding equal volume of DMSO (dimethyl sulfoxide); placing the zebra fish on a glass slide, and taking a picture of the zebra fish under a microscope and storing the zebra fish; the zebrafish trunk and thrombus-stained area (thrombus-stained area/trunk area) × 100% value was calculated using Image-Pro Plus6 as an index of the degree of thrombus generation.
TABLE 1 comparison of iron chloride groups to phenylhydrazine groups
Claims (3)
1. A method for preparing a zebra fish thrombus model by using ferric chloride is characterized by comprising the following steps:
1) selecting about 30 normal-developing 2dpf zebra fishes, transferring into 0.1-1.1% ferric chloride solution prepared from embryo culture water, culturing in an incubator for 48h, removing culture water, and dyeing in 6 micro-porous plates with o-dianisidine dyeing solution in a dark place for 15 min;
2) removing the staining solution, and rapidly washing with DMSO for 3 times;
3) transferring the zebra fish into a new 6-micropore plate, and adding equal volume of DMSO (dimethyl sulfoxide);
4) placing the zebra fish on a glass slide, and taking a picture of the zebra fish under a microscope and storing the zebra fish;
5) calculating the body part and thrombus staining area of the zebra fish by using Image-Pro Plus6, wherein (thrombus staining area/body area) × 100% is used as an index of thrombus occurrence degree;
the formula of the water for culturing the zebra fish comprises the following components: 5mmol/L NaCl, 0.17mmol/L KCl, 0.4mmol/L CaCl2，0.16mmol/L MgSO4；
The formula of the o-dianisidine staining solution comprises the following components: contains 0.6mg/mL o-dianisidine, 0.01mol/L sodium acetate, and 0.65% H2O2，0.16mmol/L MgSO4。
2. A method for preparing a zebra fish thrombus model by using ferric chloride is characterized by comprising the following steps:
1) selecting about 30 normal-developing 2dpf zebra fishes, transferring into 0.7% ferric chloride solution prepared by embryo culture water, culturing in an incubator for 48h, removing the culture water, and dyeing in 6 micro-porous plates for 15min in a dark place by using an o-dianisidine dyeing solution;
2) removing the staining solution, and rapidly washing with DMSO for 3 times;
3) transferring the zebra fish into a new 6-micropore plate, and adding equal volume of DMSO (dimethyl sulfoxide);
4) placing the zebra fish on a glass slide, and taking a picture of the zebra fish under a microscope and storing the zebra fish;
5) calculating the body part and thrombus staining area of the zebra fish by using Image-Pro Plus6, wherein (thrombus staining area/body area) × 100% is used as an index of thrombus occurrence degree;
the formula of the water for culturing the zebra fish comprises the following components: 5mmol/L NaCl, 0.17mmol/L KCl, 0.4mmol/L CaCl2，0.16mmol/L MgSO4；
The formula of the o-dianisidine staining solution comprises the following components: contains 0.6mg/mL o-dianisidine, 0.01mol/L sodium acetate, and 0.65% H2O2，0.16mmol/L MgSO4。
3. The method of using the zebrafish thrombus model of claim 2, comprising the steps of:
1) preparing a zebrafish thrombus animal model according to the method of claim 2;
2) adding the drug to be tested, and continuing culturing for a corresponding time;
3) dyeing for 15min by using an o-dianisidine dyeing solution;
4) the zebrafish trunk and thrombus staining area (thrombus staining area/trunk area) × 100% value was calculated using Image-Pro Plus6 as an index for evaluating the effect of the drug on thrombus.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004321098A (en) * | 2003-04-25 | 2004-11-18 | Fuji Xerox Co Ltd | Method for suppressing development of pigmented epithelium of fishes |
| CN102266313A (en) * | 2011-05-17 | 2011-12-07 | 杭州环特生物科技有限公司 | Method for establishing zebra fish thrombosis model and method for screening antithrombotic drug and thrombosis producing drug |
| CN102907357A (en) * | 2012-09-27 | 2013-02-06 | 杭州环特生物科技有限公司 | Building method and application of zebra fish hyperlipidemia model |
| CN106667982A (en) * | 2017-02-24 | 2017-05-17 | 北京中医药大学 | Method for preparing zebrafish thrombus model |
-
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- 2021-01-13 CN CN202110043719.3A patent/CN112715435A/en not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004321098A (en) * | 2003-04-25 | 2004-11-18 | Fuji Xerox Co Ltd | Method for suppressing development of pigmented epithelium of fishes |
| CN102266313A (en) * | 2011-05-17 | 2011-12-07 | 杭州环特生物科技有限公司 | Method for establishing zebra fish thrombosis model and method for screening antithrombotic drug and thrombosis producing drug |
| CN102907357A (en) * | 2012-09-27 | 2013-02-06 | 杭州环特生物科技有限公司 | Building method and application of zebra fish hyperlipidemia model |
| CN106667982A (en) * | 2017-02-24 | 2017-05-17 | 北京中医药大学 | Method for preparing zebrafish thrombus model |
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