MD718Z - Medium for cryopreservation of human sperm - Google Patents
Medium for cryopreservation of human sperm Download PDFInfo
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- MD718Z MD718Z MDS20130107A MDS20130107A MD718Z MD 718 Z MD718 Z MD 718Z MD S20130107 A MDS20130107 A MD S20130107A MD S20130107 A MDS20130107 A MD S20130107A MD 718 Z MD718 Z MD 718Z
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- medium
- cryopreservation
- sperm
- carnitine
- glucose
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- 238000005138 cryopreservation Methods 0.000 title claims abstract description 9
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 23
- PHIQHXFUZVPYII-ZCFIWIBFSA-N (R)-carnitine Chemical compound C[N+](C)(C)C[C@H](O)CC([O-])=O PHIQHXFUZVPYII-ZCFIWIBFSA-N 0.000 claims abstract description 12
- 102000002322 Egg Proteins Human genes 0.000 claims abstract description 11
- 108010000912 Egg Proteins Proteins 0.000 claims abstract description 11
- 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 claims abstract description 11
- 235000013345 egg yolk Nutrition 0.000 claims abstract description 11
- 239000008103 glucose Substances 0.000 claims abstract description 11
- 239000001509 sodium citrate Substances 0.000 claims abstract description 11
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 claims abstract description 11
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims abstract description 10
- 229930006000 Sucrose Natural products 0.000 claims abstract description 10
- 210000002969 egg yolk Anatomy 0.000 claims abstract description 10
- 239000005720 sucrose Substances 0.000 claims abstract description 10
- 239000012154 double-distilled water Substances 0.000 claims description 10
- 235000011187 glycerol Nutrition 0.000 claims description 10
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 4
- 238000010257 thawing Methods 0.000 abstract description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 2
- 239000000203 mixture Substances 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000001850 reproductive effect Effects 0.000 description 3
- 235000014113 dietary fatty acids Nutrition 0.000 description 2
- 229930195729 fatty acid Natural products 0.000 description 2
- 239000000194 fatty acid Substances 0.000 description 2
- 150000004665 fatty acids Chemical class 0.000 description 2
- 230000004720 fertilization Effects 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 230000019100 sperm motility Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- RGJOEKWQDUBAIZ-IBOSZNHHSA-N CoASH Chemical compound O[C@@H]1[C@H](OP(O)(O)=O)[C@@H](COP(O)(=O)OP(O)(=O)OCC(C)(C)[C@@H](O)C(=O)NCCC(=O)NCCS)O[C@H]1N1C2=NC=NC(N)=C2N=C1 RGJOEKWQDUBAIZ-IBOSZNHHSA-N 0.000 description 1
- 125000002252 acyl group Chemical group 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 210000000991 chicken egg Anatomy 0.000 description 1
- RGJOEKWQDUBAIZ-UHFFFAOYSA-N coenzime A Natural products OC1C(OP(O)(O)=O)C(COP(O)(=O)OP(O)(=O)OCC(C)(C)C(O)C(=O)NCCC(=O)NCCS)OC1N1C2=NC=NC(N)=C2N=C1 RGJOEKWQDUBAIZ-UHFFFAOYSA-N 0.000 description 1
- 239000005516 coenzyme A Substances 0.000 description 1
- 229940093530 coenzyme a Drugs 0.000 description 1
- 230000000959 cryoprotective effect Effects 0.000 description 1
- 230000000254 damaging effect Effects 0.000 description 1
- KDTSHFARGAKYJN-UHFFFAOYSA-N dephosphocoenzyme A Natural products OC1C(O)C(COP(O)(=O)OP(O)(=O)OCC(C)(C)C(O)C(=O)NCCC(=O)NCCS)OC1N1C2=NC=NC(N)=C2N=C1 KDTSHFARGAKYJN-UHFFFAOYSA-N 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 235000001727 glucose Nutrition 0.000 description 1
- 239000012456 homogeneous solution Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000002438 mitochondrial effect Effects 0.000 description 1
- 210000001700 mitochondrial membrane Anatomy 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 235000011083 sodium citrates Nutrition 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
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- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
Изобретение относится к криобиологии, а именно к среде для криоконсервации спермы человека.Среда для криоконсервации спермы человека содержит, на 100 мл: цитрат натрия 1,1…1,4 г, глюкозу 1,8…3,0 г, сахарозу 6,0…8,4 г, яичный желток 24,0…28,0 мл, глицерин 3,8…4,2 мл, L-карнитин 6,0…10,0 мг и бидистиллированную воду остальное.Результат состоит в повышении физиологических показателей спермы после оттаивания.The invention relates to cryobiology, namely, a medium for cryopreservation of human sperm. The medium for cryopreservation of human sperm contains, per 100 ml: sodium citrate 1.1 ... 1.4 g, glucose 1.8 ... 3.0 g, sucrose 6.0 ... 8.4 g, egg yolk 24.0 ... 28.0 ml, glycerol 3.8 ... 4.2 ml, L-carnitine 6.0 ... 10.0 mg and bidistilled water the rest. The result is an increase in the physiological parameters of sperm after thawing.
Description
Invenţia se referă la criobiologie, şi anume la un mediu pentru crioconservarea spermei umane. The invention relates to cryobiology, namely to a medium for cryopreservation of human sperm.
Este cunoscut mediul pentru crioconservarea spermei umane, care conţine citrat de sodiu, glucoză, gălbenuş de ou, glicerină şi apă distilată [1]. The medium for cryopreservation of human sperm is known, which contains sodium citrate, glucose, egg yolk, glycerin and distilled water [1].
Dezavantajul acestui mediu constă în aceea că el conţine doar monoglucide, ceea ce asigură o crioprotecţie mai slabă, în consecinţă indicii fiziologici ai spermei după decongelare rămân la un nivel scăzut comparativ cu sperma nativă. The disadvantage of this medium is that it contains only monoglycosides, which provides weaker cryoprotection, consequently the physiological indices of sperm after thawing remain at a low level compared to native sperm.
Cea mai apropiată soluţie este mediul pentru crioconservarea spermei umane care conţine la 100 ml: citrat de sodiu 1,1…1,4 g, glucoză 1,8…3,0 g, zaharoză 6,0…8,4 g, gălbenuş de ou 24,0…28,0 ml, glicerină 3,8…4,2 ml şi apă bidistilată restul [2]. The closest solution is the medium for cryopreservation of human sperm which contains per 100 ml: sodium citrate 1.1…1.4 g, glucose 1.8…3.0 g, sucrose 6.0…8.4 g, egg yolk 24.0…28.0 ml, glycerin 3.8…4.2 ml and the rest double-distilled water [2].
Dezavantajul acestui mediu constă în aceea că nu conţine antioxidanţi şi substanţe care ar favoriza procesele de formare a energiei, ceea ce reduce viabilitatea spermatozoizilor după decongelare. The disadvantage of this medium is that it does not contain antioxidants and substances that would favor energy formation processes, which reduces the viability of sperm after thawing.
Problema pe care o rezolvă invenţia constă în sporirea indicilor fiziologici ai spermei după decongelare. The problem that the invention solves consists in increasing the physiological indices of sperm after thawing.
Problema se soluţionează prin aceea că mediul pentru crioconservarea spermei umane conţine citrat de sodiu, glucoză, zaharoză, gălbenuş de ou, glicerină şi apă bidistilată, totodată suplimentar conţine L-carnitină, componentele fiind luate în următorul raport, la 100 ml: The problem is solved by the fact that the medium for cryopreservation of human sperm contains sodium citrate, glucose, sucrose, egg yolk, glycerin and double-distilled water, and additionally contains L-carnitine, the components being taken in the following ratio, per 100 ml:
citrat de sodiu, g 1,1…1,4 glucoză, g 1,8…3,0 zaharoză, g 6,0…8,4 L-carnitină, mg 6,0…10,0 gălbenuş de ou, ml 24,0…28,0 glicerină, ml 3,8…4,2 apă bidistilată, ml restul.sodium citrate, g 1.1…1.4 glucose, g 1.8…3.0 sucrose, g 6.0…8.4 L-carnitine, mg 6.0…10.0 egg yolk, ml 24.0…28.0 glycerin, ml 3.8…4.2 double-distilled water, ml the rest.
Rezultatul constă în sporirea indicilor fiziologici ai spermei după decongelare, menţinerea la nivel sanogen a mobilităţii, longevităţii şi indicelui absolut de supravieţuire. The result is an increase in the physiological indices of sperm after thawing, maintaining mobility, longevity and absolute survival index at a healthy level.
Rezultatul este asigurat de includerea în componenţa mediului a substanţelor se favorizează procesele de formare a energiei. The result is ensured by the inclusion in the composition of the environment of substances that favor energy formation processes.
Efectul crioprotector al mediului este asigurat de proprietatea L-carnitinei de a proteja ADN-ul şi complexul membranar al spermatozoizilor de acţiunea deterioratoare a radicalilor liberi de oxigen, precum şi de a spori permiabilitatea membranei mitocondriale pentru acizii graşi, totodată L-carnitina participă la procesele de β-oxidare a acizilor graşi, care servesc ca transportatori ai grupelor acilice spre coenzima mitocondrială A, cu formarea energiei necesare pentru menţinerea funcţiei normale a spermatozoizilor după decongelare, ceea ce asigură o capacitate sporită de fecundare a materialului reproducător decongelat. The cryoprotective effect of the environment is ensured by the property of L-carnitine to protect the DNA and membrane complex of spermatozoa from the damaging action of oxygen free radicals, as well as to increase the permeability of the mitochondrial membrane for fatty acids. At the same time, L-carnitine participates in the processes of β-oxidation of fatty acids, which serve as carriers of acyl groups to mitochondrial coenzyme A, with the formation of the energy necessary to maintain the normal function of spermatozoa after thawing, which ensures an increased fertilization capacity of the thawed reproductive material.
Conţinutul optimal al L-carnitinei în componenţa mediului elaborat a fost determinat în mod experimental. Pregătirea mediului se realizează în modul următor. Într-un vas de sticlă cotat se adaugă glucoză, zaharoză, citrat de sodiu, L-carnitină, cantităţile fiind luate în conformitate cu compoziţia propusă, şi 50 ml de apă bidistilată, se dizolvă componentele mediului, după care se mai adaugă gălbenuşul de ou de găină şi glicerina. Amestecul obţinut se agită bine până la obţinerea unei soluţii omogene şi se aduce volumul cu apă bidistilată până la 100 ml. Mediul obţinut este omogen, transparent, fără precipitat sau fulgi. Mediul se pregăteşte nemijlocit înainte de întrebuinţare. The optimal content of L-carnitine in the composition of the developed medium was determined experimentally. The preparation of the medium is carried out as follows. In a graduated glass vessel, glucose, sucrose, sodium citrate, L-carnitine are added, the quantities being taken in accordance with the proposed composition, and 50 ml of double-distilled water, the components of the medium are dissolved, after which the chicken egg yolk and glycerin are added. The resulting mixture is shaken well until a homogeneous solution is obtained and the volume is brought to 100 ml with double-distilled water. The resulting medium is homogeneous, transparent, without precipitate or flakes. The medium is prepared immediately before use.
Exemple de realizare a invenţiei Examples of embodiments of the invention
Exemplul 1 Example 1
citrat de sodiu 1,1 g glucoză 1,8 g zaharoză 6,0 g L-carnitină 6,0 mg gălbenuş de ou 24,0 ml glicerină 3,8 ml apă bidistilată restul până la 100 ml.sodium citrate 1.1 g glucose 1.8 g sucrose 6.0 g L-carnitine 6.0 mg egg yolk 24.0 ml glycerin 3.8 ml double-distilled water the rest up to 100 ml.
Exemplul 2 Example 2
citrat de sodiu 1,2 g glucoză 2,4 g zaharoză 7,2 g L-carnitină 8,0 mg gălbenuş de ou 26,0 ml glicerină 4,0 ml apă bidistilată restul până la 100 ml.sodium citrate 1.2 g glucose 2.4 g sucrose 7.2 g L-carnitine 8.0 mg egg yolk 26.0 ml glycerin 4.0 ml double distilled water the rest up to 100 ml.
Exemplul 3 Example 3
citrat de sodiu 1,4 g glucoză 3,0 g zaharoză 8,4 g L-carnitină 10,0 mg gălbenuş de ou 28,0 ml glicerină 4,2 ml apă bidistilată restul până la 100 ml.sodium citrate 1.4 g glucose 3.0 g sucrose 8.4 g L-carnitine 10.0 mg egg yolk 28.0 ml glycerin 4.2 ml double distilled water the rest up to 100 ml.
Mediul a fost experimentat în condiţii de laborator. Pentru crioconservare au fost selectate variantele de mediu conform exemplelor 1-3, testate în comparaţie cu cea mai apropiată soluţie. Rezultatele congelării spermei native în mediul din variantele propuse în invenţie sunt incluse în tab. 1. The medium was tested under laboratory conditions. For cryopreservation, the medium variants according to examples 1-3 were selected, tested in comparison with the closest solution. The results of freezing native sperm in the medium from the variants proposed in the invention are included in tab. 1.
Tabelul 1 Table 1
Indicii fiziologici Experimentele Exemplul 1 Exemplul 2 Exemplul 3 Mobilitatea, bal 4,29 ± 0,11 4,86 ± 0,09 4,35 ± 0,20 Longevitatea, ore 9,36 ± 0,20 11,80 ± 0,22 10,64 ± 0,27 Indicele absolut de supravieţuire, u.c. 119,20 ± 9,40 130,90 ± 11,30 122,80 ± 14,24Physiological indices Experiments Example 1 Example 2 Example 3 Mobility, bal 4.29 ± 0.11 4.86 ± 0.09 4.35 ± 0.20 Longevity, hours 9.36 ± 0.20 11.80 ± 0.22 10.64 ± 0.27 Absolute survival index, u.c. 119.20 ± 9.40 130.90 ± 11.30 122.80 ± 14.24
Compoziţia optimă a mediului, în care s-au manifestat cele mai pronunţate proprietăţi protectoare este cea din exemplul 2. The optimal composition of the environment, in which the most pronounced protective properties were manifested, is that in example 2.
În tab. 2 sunt prezentaţi indicii fiziologici ai spermei native şi congelate conform invenţiei şi celei mai apropiate soluţii. Table 2 presents the physiological indices of native and frozen sperm according to the invention and the closest solution.
Tabelul 2 Table 2
Indicii fiziologici Sperma nativă Soluţia cea mai apropiată Invenţia propusă Mobilitatea, bal 7,00 ± 0,01 4,59 ± 0,14 4,50 ± 0,13 Longevitatea, ore 21,00 ± 3,26 9,30 ± 0,22 10,60 ± 0,23 Indicele absolut de supravieţuire, u.c. 430,00 ± 41,80 67,60 ± 9,45 124,3 ± 11,64Physiological indices Native sperm Closest solution Proposed invention Motility, bac 7.00 ± 0.01 4.59 ± 0.14 4.50 ± 0.13 Longevity, hours 21.00 ± 3.26 9.30 ± 0.22 10.60 ± 0.23 Absolute survival index, u.c. 430.00 ± 41.80 67.60 ± 9.45 124.3 ± 11.64
Analiza rezultatelor obţinute denotă despre eficienţa mediului propus, comparativ cu cea mai apropiată soluţie. The analysis of the results obtained indicates the efficiency of the proposed environment, compared to the closest solution.
Mediul propus permite de a menţine indicii mobilităţii şi longevităţii spermatozoizilor la un nivel funcţional înalt după decongelare. Mobilitatea spermatozoizilor după decongelare constituie 63,5%, longevitatea 52,7%, indicele absolut de supravieţuire 28,9% comparativ cu sperma nativă, ceea ce asigură o capacitate sporită de fecundare a materialului reproducător decongelat. The proposed medium allows maintaining sperm motility and longevity indices at a high functional level after thawing. Sperm motility after thawing is 63.5%, longevity 52.7%, absolute survival index 28.9% compared to native sperm, which ensures an increased fertilization capacity of thawed reproductive material.
Mediul propus asigură o calitate eficientă a materialului reproducător decongelat. The proposed environment ensures efficient quality of the thawed reproductive material.
1. Грищенко В. И., Дунаевская А. В., Калугин Ю. С. Влияние быстрых и сверхбыстрых скоростей замораживания на сохранность спермиев человека. Проблемы криобиологии, 2000, № 2, с. 53. 1. Grishchenko V. I., Dunaevskaya A. V., Kalugin Yu. С. The influence of fast and superfast freezing speeds on the safety of human sperm. Проблемы криобиологии, 2000, № 2, с. 53.
2. MD 201 Y 2010.05.31 2. MD 201 Y 2010.05.31
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| Application Number | Priority Date | Filing Date | Title |
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| MDS20130107A MD718Z (en) | 2013-06-14 | 2013-06-14 | Medium for cryopreservation of human sperm |
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| MDS20130107A MD718Z (en) | 2013-06-14 | 2013-06-14 | Medium for cryopreservation of human sperm |
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| MD718Y MD718Y (en) | 2014-01-31 |
| MD718Z true MD718Z (en) | 2014-08-31 |
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| MDS20130107A MD718Z (en) | 2013-06-14 | 2013-06-14 | Medium for cryopreservation of human sperm |
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| CN113186101B (en) * | 2021-04-15 | 2022-10-14 | 上海理工大学 | Low-temperature protective agent, preparation method and preservation method |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SU1660651A1 (en) * | 1985-04-18 | 1991-07-07 | Институт проблем криобиологии и криомедицины АН УССР | Method for human sperm conservation |
| RU2237712C2 (en) * | 2002-05-06 | 2004-10-10 | Кострикин Александр Александрович | Medium for cryopreserving human and animal cells |
| RU2257710C2 (en) * | 2003-10-28 | 2005-08-10 | Одинцов Андрей Александрович | Method for cryopreserving human sperm |
| MD3035G2 (en) * | 2005-07-18 | 2006-10-31 | Национальный Институт Зоотехнии И Ветеринарии | Medium for boar semen dilution |
| MD58Z (en) * | 2009-03-13 | 2010-03-31 | Институт Физиологии И Санокреатологии Академии Наук Молдовы | Human semen cryoconservation medium |
| MD201Y (en) * | 2010-02-11 | 2010-05-31 | Institutul De Fiziologie Si Sanocreatologie Al Academiei De Stiinte A Moldovei | Medium for cryoconservation of human sperm |
| EA015422B1 (en) * | 2006-02-03 | 2011-08-30 | Фапа Витал Анштальт | Combination preparation for improving sperm quality |
-
2013
- 2013-06-14 MD MDS20130107A patent/MD718Z/en not_active IP Right Cessation
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SU1660651A1 (en) * | 1985-04-18 | 1991-07-07 | Институт проблем криобиологии и криомедицины АН УССР | Method for human sperm conservation |
| RU2237712C2 (en) * | 2002-05-06 | 2004-10-10 | Кострикин Александр Александрович | Medium for cryopreserving human and animal cells |
| RU2257710C2 (en) * | 2003-10-28 | 2005-08-10 | Одинцов Андрей Александрович | Method for cryopreserving human sperm |
| MD3035G2 (en) * | 2005-07-18 | 2006-10-31 | Национальный Институт Зоотехнии И Ветеринарии | Medium for boar semen dilution |
| EA015422B1 (en) * | 2006-02-03 | 2011-08-30 | Фапа Витал Анштальт | Combination preparation for improving sperm quality |
| MD58Z (en) * | 2009-03-13 | 2010-03-31 | Институт Физиологии И Санокреатологии Академии Наук Молдовы | Human semen cryoconservation medium |
| MD201Y (en) * | 2010-02-11 | 2010-05-31 | Institutul De Fiziologie Si Sanocreatologie Al Academiei De Stiinte A Moldovei | Medium for cryoconservation of human sperm |
Non-Patent Citations (1)
| Title |
|---|
| Грищенко В. И., Дунаевская А. В., Калугин Ю. С. Влияние быстрых и сверхбыстрых скоростей замораживания на сохранность спермиев человека. Проблемы криобиологии, 2000, № 2, с. 53. * |
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| Publication number | Publication date |
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| MD718Y (en) | 2014-01-31 |
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