CN101622986A - Glutathione-contained boar semen cryopreservation liquid and cryopreservation method thereof - Google Patents
Glutathione-contained boar semen cryopreservation liquid and cryopreservation method thereof Download PDFInfo
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- CN101622986A CN101622986A CN200910100965A CN200910100965A CN101622986A CN 101622986 A CN101622986 A CN 101622986A CN 200910100965 A CN200910100965 A CN 200910100965A CN 200910100965 A CN200910100965 A CN 200910100965A CN 101622986 A CN101622986 A CN 101622986A
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- 238000005138 cryopreservation Methods 0.000 title claims abstract description 60
- 239000007788 liquid Substances 0.000 title claims abstract description 45
- 210000000582 semen Anatomy 0.000 title claims abstract description 34
- 238000000034 method Methods 0.000 title claims abstract description 17
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 claims abstract description 90
- 108010024636 Glutathione Proteins 0.000 claims abstract description 45
- 229960003180 glutathione Drugs 0.000 claims abstract description 45
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 32
- 235000011187 glycerol Nutrition 0.000 claims abstract description 16
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims abstract description 15
- 239000008101 lactose Substances 0.000 claims abstract description 15
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 claims abstract description 14
- 235000019333 sodium laurylsulphate Nutrition 0.000 claims abstract description 14
- 238000010790 dilution Methods 0.000 claims description 42
- 239000012895 dilution Substances 0.000 claims description 42
- 238000007710 freezing Methods 0.000 claims description 17
- 230000008014 freezing Effects 0.000 claims description 17
- 210000002969 egg yolk Anatomy 0.000 claims description 14
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 12
- 238000002360 preparation method Methods 0.000 claims description 7
- 229910052757 nitrogen Inorganic materials 0.000 claims description 6
- 238000004321 preservation Methods 0.000 claims description 5
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- 238000001556 precipitation Methods 0.000 claims description 3
- 238000007865 diluting Methods 0.000 claims description 2
- 239000006228 supernatant Substances 0.000 claims description 2
- 230000008569 process Effects 0.000 abstract description 7
- 239000000758 substrate Substances 0.000 abstract description 3
- 230000004083 survival effect Effects 0.000 abstract description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- 239000003085 diluting agent Substances 0.000 abstract 1
- 238000005057 refrigeration Methods 0.000 abstract 1
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 8
- 150000003254 radicals Chemical class 0.000 description 7
- 102000004190 Enzymes Human genes 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 6
- 230000004899 motility Effects 0.000 description 5
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 4
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 4
- 239000003963 antioxidant agent Substances 0.000 description 4
- 235000006708 antioxidants Nutrition 0.000 description 4
- 238000009395 breeding Methods 0.000 description 4
- 230000001488 breeding effect Effects 0.000 description 4
- 244000144972 livestock Species 0.000 description 4
- 230000019100 sperm motility Effects 0.000 description 4
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 4
- 230000003064 anti-oxidating effect Effects 0.000 description 3
- 230000003078 antioxidant effect Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 102000006587 Glutathione peroxidase Human genes 0.000 description 2
- 108700016172 Glutathione peroxidases Proteins 0.000 description 2
- 108010070675 Glutathione transferase Proteins 0.000 description 2
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- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 229930182555 Penicillin Natural products 0.000 description 2
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 2
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
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- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention provides glutathione-contained boar semen cryopreservation liquid and a cryopreservation method thereof. The boar semen cryopreservation liquid uses diluent which is generally suitable for boar semen cryopreservation as a substrate and also comprises 2-6 mg/ml of glutathione, 5-11 mg/ml of lactose, 10-30 percent (v/v) of yelk, 5-10 percent (v/v) of glycerin and 0.5-2 mg/ml of sodium lauryl sulfate. The invention applies the glutathione to the boar semen cryopreservation, can effectively remove redundant free radicals generated in a refrigeration process, solves the problem of low survival rate of the prior boar semen cryopreservation, has lower economic cost and is convenient to popularize and apply in production.
Description
(1) technical field
The present invention relates to a kind of Cryopreservation of Boar Semen liquid and freezing and storing method thereof that contains glutathione, belong to the livestock reproduction technical field.
(2) background technology
In China, pig is topmost stock raising, and pork output accounts for 70% of national meat total amount.Along with the raising of living standards of the people, the breeding direction of pig improved the local flavor of meat from mainly being conceived to the raising of growth rate and lean meat percentage, changing into improve pork quality in recent years.Traditional seed selection mode time generation interval is long, can not meeting the market requirement, and adopt pig frozen semen technology, can make full use of good breeding stock, shorten time fine-variety breeding generation interval, set up the boar and the market pig crossbreed system of new suitable market demands fast.Simultaneously, the raising history of nearly one thousand years, formed hundreds of landrace that differ from one another in China, this is a very valuable property, the competition of livestock breeding from now on, it quite partly is genetic resources, the competition that excellent genes utilizes, but the introduction of external pig kind hybridization have in recent years made China that a considerable amount of landrace extinctions are arranged or have faced extinction, and adopt the Cryopreservation of Boar Semen technology can effectively preserve China's local pig breed matter resource, have immeasurable social effect and benefit.
Though the freezing preservation technology of herbvore livestock semen ultralow temperature (196 ℃) is being brought into play important effect in preserving, and is being produced obvious economic and social benefit in modernized herding production, germ plasm resource.But the pig frozen semen is difficult to apply on a large scale aborning always, its reason has two, the one, C/P ratio significantly is lower than plant-eating animal in the pig spermoblast plasma membrane, the ion permeability of sperm membrane strengthens in refrigerating process, lipid material runs off, and causes freezing that smart motility rate is low, conception rate is low.The 2nd, the pig sperm often is subjected to the infringement of free radical and oxidative stress in refrigerating process, thereby influences motility rate and fertility behind the pig sperm freezing.Free radical is that eubolism produces in the cell, organism contain natural antioxidant (free radical scavenger) with in and the toxic and side effect of free radical.Under the adjusting of these antioxidants, the generation of interior free yl and elimination are in a kind of dynamic equilibrium just.But, the level of above-mentioned antioxidant is lower than level in the body in the boar semen dilution of routine, and in the dilution equilibrium process before sperm freezing, be exposed to air inevitably, and airborne oxygen concentration is higher than oxygen concentration (20%VS.7%) in the genital tract, and the influence of metal ion in light and the dilution when adding manipulation in vitro makes when freezing that the free radical level increases in the sperm, break the poised state of oxidation antioxidation, caused the damage of sperm oxidative stress.Therefore, the new cryoprotector of screening has become the focus of boar semen research field to improve freezing back sperm motility rate.
Glutathione is the non-albumen tripeptides that contains sulfydryl that methionine, glycine and cysteine are formed, and has stronger antioxidation, can eliminate free radicals such as superoxide anion and hydrogen peroxide.It is a non-albumen sulfhydryl compound main in the histocyte, is the substrate of glutathione peroxidase and two kinds of enzymes of glutathione S-transferase, and is essential by these two kinds of enzyme decomposition of hydrogen peroxide, can stablize the enzyme that contains sulfydryl.For the existing bibliographical information of the research of glutathione.Multiple patent applications such as CN100455592C, CN100503837C, CN1164737C, CN1325511C, CN1313597C, CN100360679C disclose the preparation method of glutathione.Multiple patent disclosure such as CN1185008C, CN100435793C glutathione as the purposes of medicine.The relevant glutathione of Shang Weijian is used for the livestock reproduction technical field, especially for the report of Cryopreservation of Boar Semen but up to the present.
(3) summary of the invention
The purpose of this invention is to provide a kind of Cryopreservation of Boar Semen liquid and using method thereof of containing glutathione that reduces freezing damage, improves cold smart motility rate to sperm.
The technical solution used in the present invention is:
The application of glutathione in preparation Cryopreservation of Boar Semen liquid.Concrete, described Cryopreservation of Boar Semen liquid is matrix with the dilution that is applicable to Cryopreservation of Boar Semen, the glutathione that also contains 2～6mg/mL in the described Cryopreservation of Boar Semen liquid, the lactose of 5～11mg/mL and 10～30% (v/v) yolk, the glycerine of 5～10% (v/v) and the sodium lauryl sulfate of 0.5～2mg/mL.A kind of Cryopreservation of Boar Semen liquid that contains glutathione, described Cryopreservation of Boar Semen liquid is applicable to that with routine the dilution of Cryopreservation of Boar Semen is a matrix, the glutathione that also contains 2～6mg/mL in the described Cryopreservation of Boar Semen liquid, the lactose of 5～11mg/mL and 10～30% (v/v) yolk, the glycerine of 5～10% (v/v) and the sodium lauryl sulfate of 0.5～2mg/mL.
Described glutathione has multiple preparation method, belong to already known processes and mature technology, and existing commodity selling, comparatively Cheng Shu manufacturer has U.S. Sigma-Aldrich company, Shaanxi to melt to rise bio tech ltd, Chongqing Yaoyou Pharmaceutical Co., Ltd., Huzhou is bright is Bioisystech Co., Ltd etc. deeply.Used glutathione is provided by U.S. Sigma-Aldrich company among the present invention, and purity is 98%.
Concrete, every 1000mL is composed as follows for described Cryopreservation of Boar Semen liquid:
Glutathione 2～6g
Lactose 50～110g
Yolk 100～300mL
Glycerine 50～100mL
Sodium lauryl sulfate 5～20g
The dilution that is applicable to Cryopreservation of Boar Semen complements to 1000mL.
The described dilution of Cryopreservation of Boar Semen that is applicable to is the conventional dilution that is used for Cryopreservation of Boar Semen, as the Seminark of BTS dilution, Korea S's product
TMDilution etc.Preferably, describedly be applicable to that the dilution of Cryopreservation of Boar Semen is BTS dilution (Beltsville thawingsolution), every 1000mL presses following formulated: glucose 37g, sodium citrate 6g, potassium chloride 0.75g, sodium bicarbonate 1.25g, EDTA 1.25g, penicillin 100000 units, streptomycin 100mg, deionized water complements to 1000mL.
Preferably, the every 1000mL of described Cryopreservation of Boar Semen liquid is composed as follows:
Glutathione 4g
Lactose 80g
Yolk 200mL
Glycerine 80mL
Sodium lauryl sulfate 5g
The BTS dilution complements to 1000mL.
The invention still further relates to a kind of method that the described Cryopreservation of Boar Semen liquid that contains glutathione carries out Cryopreservation of Boar Semen of using, described method comprises:
(1) with the fresh pig seminal fluid taked with one times of the BTS diluted of isothermal;
(2) left standstill (balance) 2 hours under 15 ℃ of conditions, 1000 left the heart 10 minutes;
(3) supernatant is removed in centrifugal back, the precipitation boar semen diluted (not containing glycerine and sodium lauryl sulfate) that contains glutathione, and making sperm concentration is 2 * 10
9Individual/mL, the seminal fluid after the dilution leaves standstill (balance) after 2 hours under 5 ℃ of conditions, and with one times of 5 ℃ of Cryopreservation of Boar Semen liquid dilution that contain glutathione of preserving down, the final densities that makes sperm is 1 * 10 again
9Individual/mL, the seminal fluid after obtaining diluting;
Every 1000mL is composed as follows for described boar semen dilution:
Glutathione 2～6g
Lactose 50～110g
Yolk 100～300mL
The BTS dilution complements to 1000mL;
Every 1000mL is composed as follows for described Cryopreservation of Boar Semen liquid:
Glutathione 2～6g
Lactose 50～110g
Yolk 100～300mL
Glycerine 50～100mL
Sodium lauryl sulfate 5～20g
The BTS dilution complements to 1000mL;
(4) seminal fluid after the chill back is packed into conventional freezing pipe places on the liquid nitrogen surface 3 centimetres,
Fumigate after 5 minutes, directly drop in the liquid nitrogen, freezing preservation is standby.
Cryopreservation of Boar Semen liquid of the present invention is according to needs anti-oxidant in the Cryopreservation of Boar Semen process, Green Tea Extract, develops in conjunction with the biological function of glutathione.Glutathione has stronger antioxidation, can eliminate free radicals such as superoxide anion and hydrogen peroxide.It is a non-albumen sulfhydryl compound main in the histocyte, is the substrate of glutathione peroxidase and two kinds of enzymes of glutathione S-transferase, and is essential by these two kinds of enzyme decomposition of hydrogen peroxide, can stablize the enzyme that contains sulfydryl.Therefore glutathione is applied to Cryopreservation of Boar Semen, can remove the unnecessary free radical that produces in the refrigerating process effectively, has solved the low problem of present Cryopreservation of Boar Semen survival rate.Its Financial cost is lower, is convenient to apply aborning.
(4) embodiment
The present invention is described further below in conjunction with specific embodiment, but protection scope of the present invention is not limited in this:
Embodiment 1: preparation contains the Cryopreservation of Boar Semen liquid and the freeze-extender of glutathione
Contain the Cryopreservation of Boar Semen liquid preparation of glutathione:
Accurately take by weighing glutathione 400mg, (every 1000mL presses following formulated to first BTS dilution with 50mL: glucose 37g, sodium citrate 6g, potassium chloride 0.75g, sodium bicarbonate 1.25g, EDTA 1.25g, penicillin 100000 units, streptomycin 100mg, deionized water complements to 1000mL) dissolving, the sodium lauryl sulfate that in dilution, adds yolk (from egg, extract and obtain), 8mL glycerine and the 0.5g of 8g lactose, 20mL then, add the BTS dilution again to 100mL, once more stirring and evenly mixing.After the micro-filtration membrane suction filtration sterilization with 0.22 μ M, store down at 5 ℃, standby.
Contain the boar semen dilution preparation of glutathione:
Accurately take by weighing glutathione 400mg,, in dilution, add the yolk of 8g lactose and 20mL then, add the BTS dilution again to 100mL, once more stirring and evenly mixing earlier with the BTS dilution dissolving of 50mL.After the micro-filtration membrane suction filtration sterilization with 0.22 μ M, store down at 5 ℃, standby.
Embodiment 2: the Cryopreservation of Boar Semen liquid that contains glutathione is used for big York Cryopreservation of Boar Semen
Select fertility height, athletic big York stock boar, adopt the hand grip semen collection.Collect sperm and enrich part, through 4 layers of sterile gauze filter remove colloid after, survey sperm volume immediately, and check sperm motility rate, collect sperm motility rate at the about 50mL of the seminal fluid 75% or more, use one times of the BTS diluted of isothermal.Under 15 ℃ of conditions, leave standstill balance 2 hours, 1000 left the heart 10 minutes then.
Seminal plasma is removed in the centrifugal back of seminal fluid, the sperm precipitation is diluted with the boar semen dilution (embodiment 1) that contains glutathione, this dilution is the freezing preservation liquid of the present invention that does not contain glycerine and sodium lauryl sulfate, because under the glycerine cryogenic conditions to the sperm toxic side effect, add behind the glycerine sperm want at once freezing can not stand at low temperature.Making sperm concentration is 2 * 10
9Individual/mL.Seminal fluid after the dilution left standstill balance 2 hours under 5 ℃ of conditions.Use one times of Cryopreservation of Boar Semen liquid (embodiment 1) dilution again, the final densities that makes sperm is 1 * 10
9Individual/mL.The control group seminal fluid is with freezing preservation liquid (BTS dilution) dilution that does not contain glutathione, in the 0.25mL conventional freezing pipe of then experimental group and control group seminal fluid being packed into, places on the liquid nitrogen surface 3 centimetres, and is stifling after 5 minutes, drops into directly that liquid nitrogen is medium-term and long-term to be preserved.
Embodiment 3: big yorker is frozen essence and thaws
In order to detect the implementation result of patent of the present invention, after big yorker seminal fluid is handled according to embodiment 1, embodiment 2 methods successively, the experimental group and the control group cryovial that freeze essence will be housed respectively places 37 ℃ of water-baths to thaw in 10 seconds, microscopically detects the energy motility rate, wherein control group freeze essence thaw the back motility rate be 30.2%, freezing the essence back sperm motility rate of thawing in the experimental group cryovial is 36.7%, improves 6.5% than control group, both significant differences of statistical results show.The result confirms, the Cryopreservation of Boar Semen liquid cooling that contains glutathione is frozen and preserved big yorker seminal fluid and can effectively improve survival rate after cold essence is thawed.
Claims (7)
1. the application of glutathione in preparation Cryopreservation of Boar Semen liquid.
2. application as claimed in claim 1, it is characterized in that described Cryopreservation of Boar Semen liquid is matrix with the dilution that is applicable to Cryopreservation of Boar Semen, described Cryopreservation of Boar Semen liquid also contains the glutathione of 2～6mg/mL, the lactose of 5～11mg/mL and 10～30% yolk, 5～10% glycerine and the sodium lauryl sulfate of 0.5～2mg/mL.
3. Cryopreservation of Boar Semen liquid that contains glutathione, it is characterized in that described Cryopreservation of Boar Semen liquid is matrix with the dilution that is applicable to Cryopreservation of Boar Semen, the glutathione that also contains 2～6mg/mL in the described Cryopreservation of Boar Semen liquid, the lactose of 5～11mg/mL and 10～30% yolk, 5～10% glycerine and the sodium lauryl sulfate of 0.5～2mg/mL.
4. Cryopreservation of Boar Semen liquid as claimed in claim 3 is characterized in that the every 1000mL of described Cryopreservation of Boar Semen liquid is composed as follows:
Glutathione 2～6g
Lactose 50～110g
Yolk 100～300mL
Glycerine 50～100mL
Sodium lauryl sulfate 5～20g
The dilution that is applicable to Cryopreservation of Boar Semen complements to 1000mL.
5. as claim 3 or 4 described Cryopreservation of Boar Semen liquid, it is characterized in that describedly being applicable to that the dilution of Cryopreservation of Boar Semen is the BTS dilution.
6. Cryopreservation of Boar Semen liquid as claimed in claim 3 is characterized in that the every 1000mL of described Cryopreservation of Boar Semen liquid is composed as follows:
Glutathione 4g
Lactose 80g
Yolk 200mL
Glycerine 80mL
Sodium lauryl sulfate 5g
The BTS dilution complements to 1000mL.
7. use the method that Cryopreservation of Boar Semen liquid as claimed in claim 3 carries out Cryopreservation of Boar Semen for one kind, described method comprises:
(1) with the fresh pig seminal fluid taked with one times of the BTS diluted of isothermal;
(2) left standstill 2 hours under 15 ℃ of conditions, 1000 left the heart 10 minutes;
(3) supernatant is removed in centrifugal back, the precipitation boar semen diluted that contains glutathione, and making sperm concentration is 2 * 10
9Individual/mL, the seminal fluid after the dilution is after leaving standstill 2 hours under 5 ℃ of conditions, and with one times of 5 ℃ of described Cryopreservation of Boar Semen liquid dilution of preserving down, the final densities that makes sperm is 1 * 10 again
9Individual/mL, the seminal fluid after obtaining diluting;
The described every 1000mL of boar semen dilution that contains glutathione is composed as follows:
Glutathione 2～6g
Lactose 50～110g
Yolk 100～300mL
The BTS dilution complements to 1000mL;
Every 1000mL is composed as follows for described Cryopreservation of Boar Semen liquid:
Glutathione 2～6g
Lactose 50～110g
Yolk 100～300mL
Glycerine 50～100mL
Sodium lauryl sulfate 5～20g
The BTS dilution complements to 1000mL;
(4) seminal fluid after the chill back is packed into cryovial places on the liquid nitrogen surface 3 centimetres, stiflingly directly drops in the liquid nitrogen after 5 minutes, and freezing preservation is standby.
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| CN2009101009657A CN101622986B (en) | 2009-08-10 | 2009-08-10 | Glutathione-contained boar semen cryopreservation liquid and cryopreservation method thereof |
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| Application Number | Title | Priority Date | Filing Date |
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| CN2012101003455A Division CN102630665A (en) | 2009-08-10 | 2009-08-10 | Method for freezing and preserving pig semen |
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| CN101622986B CN101622986B (en) | 2012-11-14 |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102258007A (en) * | 2011-09-02 | 2011-11-30 | 大连海洋大学 | Refrigerant and method for storing Pacific codfish sperms |
| CN103314948A (en) * | 2013-05-31 | 2013-09-25 | 中国科学院海洋研究所 | Superfreeze preservation method of starry flounder sperms |
| CN103651330A (en) * | 2012-09-21 | 2014-03-26 | 临沂思科生物科技有限公司 | Animal semen preservative with biological activity and preparation method thereof |
| WO2020207150A1 (en) * | 2019-04-09 | 2020-10-15 | 中国科学院化学研究所 | Biomimetic ice-inhibiting material and cryopreservation liquid containing same |
| CN113331178A (en) * | 2021-06-01 | 2021-09-03 | 西北农林科技大学 | Application of methionine in preparation of diluent for preserving boar semen at normal temperature |
-
2009
- 2009-08-10 CN CN2009101009657A patent/CN101622986B/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102258007A (en) * | 2011-09-02 | 2011-11-30 | 大连海洋大学 | Refrigerant and method for storing Pacific codfish sperms |
| CN103651330A (en) * | 2012-09-21 | 2014-03-26 | 临沂思科生物科技有限公司 | Animal semen preservative with biological activity and preparation method thereof |
| CN103314948A (en) * | 2013-05-31 | 2013-09-25 | 中国科学院海洋研究所 | Superfreeze preservation method of starry flounder sperms |
| CN103314948B (en) * | 2013-05-31 | 2014-07-23 | 中国科学院海洋研究所 | Superfreeze preservation method of starry flounder sperms |
| WO2020207150A1 (en) * | 2019-04-09 | 2020-10-15 | 中国科学院化学研究所 | Biomimetic ice-inhibiting material and cryopreservation liquid containing same |
| AU2020256938B2 (en) * | 2019-04-09 | 2023-12-07 | Institute Of Chemistry, Chinese Academy Of Sciences | Biomimetic ice-inhibiting material and cryopreservation liquid containing same |
| CN113331178A (en) * | 2021-06-01 | 2021-09-03 | 西北农林科技大学 | Application of methionine in preparation of diluent for preserving boar semen at normal temperature |
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| CN101622986B (en) | 2012-11-14 |
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