Effects of L-carnitine and cryodevices on the vitrification and developmental competence of invitro fertilized buffalo oocytes

Published: Jul 9, 2022
L carnitine buffalo oocytes Straw Open pulled-straw Vitrification
Mohammed A Elmetwally
Amira M Mostagir
Fuller W Bazer
Abdelmonem Montaser
Magdy Badr
Wael Eldomany
Ashraf Eldesoky
Mona M Elghareeb
Daniela Jaimes
Yaser Y Lenis
Samy M Zaabel

In the current, study the effect of the addition of L-carnitine (LC) in in vitro maturation (IVM) medium for buffalo oocytes and different cryo-devices on developmental competence. They were matured in IVM medium supplemented with 0.0, 0.3, 0.6 and 1.2 mg/mL of LC and vitrified by using either straw (ST), open pulled-straw (OPS), solid surface vitrification (SSV). The effects of LC during invitro fertilization and invitro culture on the developmental potential were examined. ST showed a higher recovery rate when using 0.6 mg/ml; viability rate (VR) when using 0.3mg/ml and lower zona pellucida (ZP) and cytoplasmic abnormalities when using 1.2 mg/ml LC (P<0.05). OPS showed a higher recovery and viability rates when using 0.3 mg (P<0.05). SSD showed a higher recovery and VR when using 0.6 mg/ml LC (P<0.05). Maturation and fertilization rates were increased when using 0.3 mg LC and OPS (P<0.05). Cleavage was increased in 0.3 (OPS) and 0.6 mg LC (SSD). The blastocyst rate was increased in 0.3 (OPS) and 0.6 mg/ml (SSD) (P< 0.05). In conclusion, 0.3 mg/ml LC and OPS gives a higher recovery and viability rates but 0.6 mg/ml LC and of both SSD and ST gives a higher recovery and viability rates.

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Author Biography
Mohammed A Elmetwally, Associate Professor of Animal Reproduction, Faculty of Veterinary Medicine, Mansoura University, Egypt
Theriogenology Department
Arav A, Natan Y (2019) The Near Future of Vitrification of Oocytes and Embryos: Looking into Past Experience and Planning into the Future. Transfus Med Hemother 46:182–187. doi: 10.1159/000497749
Arav A, Zeron Y (1997) Vitrification of bovine oocytes using modified minimum drop size technique (MDS) is effected by the composition and the concentration of the vitrification solution and by the cooling conditions | Semantic Scholar.
Asgari V, Hosseini SM, Ostadhosseini S, et al (2012) Specific activation requirements of in vitro-matured sheep oocytes following vitrification-warming. Mol Reprod Dev 79:434–444. doi: 10.1002/mrd.22047
Attanasio L, Boccia L, Vajta G, et al (2010) Cryotop vitrification of buffalo (Bubalus bubalis) in vitro matured oocytes: effects of cryoprotectant concentrations and warming procedures. Reprod Domest Anim 45:997–1002. doi: 10.1111/j.1439-0531.2009.01475.x
Bartolac LK, Lowe JL, Koustas G, et al (2018) Effect of different penetrating and non-penetrating cryoprotectants and media temperature on the cryosurvival of vitrified in vitro produced porcine blastocysts. Anim Sci J 89:1230–1239. doi: 10.1111/asj.12996
Berkane N, Liere P, Oudinet J-P, et al (2017) From pregnancy to preeclampsia: A key role for estrogens. Endocr Rev 38:123–144. doi: 10.1210/er.2016-1065
Bus A, Langbeen A, Martin B, et al (2019) Is the pre-antral ovarian follicle the ’holy grail’for female fertility preservation? Anim Reprod Sci 207:119–130. doi: 10.1016/j.anireprosci.2019.05.017
Busardò FP, Vergallo GM, Turillazzi E, et al (2016) Accidental Thawing of Embryos, Cryopreserved for Transfer. Two Italian cases, Milan and Rome. Curr Pharm Biotechnol 17:321–325. doi: 10.2174/1389201017666151231095901
Chang H, Chen H, Zhang L, et al (2019) Effect of oocyte vitrification on DNA damage in metaphase II oocytes and the resulting preimplantation embryos. Mol Reprod Dev 86:1603–1614. doi: 10.1002/mrd.23247
Chankitisakul V, Somfai T, Inaba Y, et al (2013) Supplementation of maturation medium with L-carnitine improves cryo-tolerance of bovine in vitro matured oocytes. Theriogenology 79:590–598. doi: 10.1016/j.theriogenology.2012.11.011
Choudhary KK, Kavya KM, Jerome A, Sharma RK (2016) Advances in reproductive biotechnologies. Vet World 9:388–395. doi: 10.14202/vetworld.2016.388-395
Chung J-T, Tosca L, Huang T-H, et al (2007) Effect of polyvinylpyrrolidone on bovine oocyte maturation in vitro and subsequent fertilization and embryonic development. Reprod Biomed Online 15:198–207. doi: 10.1016/s1472-6483(10)60709-2
Cohen M, Bischof P (2007) Factors regulating trophoblast invasion. Gynecol Obstet Invest 64:126–130. doi: 10.1159/000101734
Cuello C, Sanchez-Osorio J, Almiñana C, et al (2010) Superfine open pulled straws vitrification of porcine blastocysts does not require pretreatment with cytochalasin B and/or centrifugation. Reprod Fertil Dev 22:808–817. doi: 10.1071/RD09160
El-Shalofy AS, Moawad AR, Darwish GM, et al (2017) Effect of different vitrification solutions and cryodevices on viability and subsequent development of buffalo oocytes vitrified at the germinal vesicle (GV) stage. Cryobiology 74:86–92. doi: 10.1016/j.cryobiol.2016.11.010
Elmetwally MA, Lenis Y, Tang W, et al (2018) Effects of catecholamines on secretion of interferon tau and expression of genes for synthesis of polyamines and apoptosis by ovine trophectoderm. Biol Reprod 99:611–628. doi: 10.1093/biolre/ioy085
Elmetwally M, Rohn K, Meinecke-Tillmann S (2016) Noninvasive color Doppler sonography of uterine blood flow throughout pregnancy in sheep and goats. Theriogenology 85:1070–9.e1. doi: 10.1016/j.theriogenology.2015.11.018
Gautam SK, Verma V, Palta P, et al (2008) Effect of type of cryoprotectant on morphology and developmental competence of in vitro-matured buffalo (Bubalus bubalis) oocytes subjected to slow freezing or vitrification. Reprod Fertil Dev 20:490–496. doi: 10.1071/rd07203
Gohar MA, Elmetwally MA, Montaser A, Zaabel SM (2018) Effect of Oxytetracycline Treatment on Postpartum Reproductive Performance in Dairy Buffalo-Cows with Retained Placenta in Egypt. JVHC 1:45–53. doi: 10.14302/issn.2575-1212.jvhc-18-2146
Gupta MK, Uhm SJ, Lee HT (2007) Cryopreservation of immature and in vitro matured porcine oocytes by solid surface vitrification. Theriogenology 67:238–248. doi: 10.1016/j.theriogenology.2006.07.015
Gutnisky C, Morado S, Gadze T, et al (2019) Morphological, biochemical and functional studies to evaluate bovine oocyte vitrification. Theriogenology 143:18–26. doi: 10.1016/j.theriogenology.2019.11.037
Hatırnaz Ş, Ata B, Hatırnaz ES, et al (2018) Oocyte in vitro maturation: A sytematic review. Turk J Obstet Gynecol 15:112–125. doi: 10.4274/tjod.23911
Huang JYJ, Chen HY, Park JYS, et al (2008) Comparison of spindle and chromosome configuration in in vitro- and in vivo-matured mouse oocytes after vitrification. Fertil Steril 90:1424–1432. doi: 10.1016/j.fertnstert.2007.07.1335
Huang Z, Liu J, Gao L, et al (2019) The impacts of laser zona thinning on hatching and implantation of vitrified-warmed mouse embryos. Lasers Med Sci 34:939–945. doi: 10.1007/s10103-018-2681-8
Isachenko V, Isachenko E, Michelmann HW, et al (2001) Lipolysis and ultrastructural changes of intracellular lipid vesicles after cooling of bovine and porcine GV-oocytes. Anat Histol Embryol 30:333–338. doi: 10.1046/j.1439-0264.2001.00339.x
Jain JK, Paulson RJ (2006) Oocyte cryopreservation. Fertil Steril 86:1037–1046. doi: 10.1016/j.fertnstert.2006.07.1478
Jiang Y, Han W, Shen T, Wang M-H (2012) Antioxidant Activity and Protection from DNA Damage by Water Extract from Pine (Pinus densiflora) Bark. Prev Nutr Food Sci 17:116–121. doi: 10.3746/pnf.2012.17.2.116
Kelly J, Kleemann D, Kuwayama M, Walker S (2006) 99 effect of cysteamine on survival of bovine and ovine oocytes vitrified using the minimum volume cooling (mvc) cryotop method. Reprod Fertil Dev 18:158. doi: 10.1071/RDv18n2Ab99
Kerner J, Hoppel C (2000) Fatty acid import into mitochondria. Biochim Biophys Acta 1486:1–17. doi: 10.1016/s1388-1981(00)00044-5
Khalili MA, Shahedi A, Ashourzadeh S, et al (2017) Vitrification of human immature oocytes before and after in vitro maturation: a review. J Assist Reprod Genet 34:1413–1426. doi: 10.1007/s10815-017-1005-4
Lahneche AM, Boucheham R, Ozen T, et al (2019) In vitro antioxidant, DNA-damaged protection and antiproliferative activities of ethyl acetate and n-butanol extracts of Centaurea sphaerocephalaL. An Acad Bras Cienc 91:e20180462. doi: 10.1590/0001-3765201920180462
Ledda S, Bogliolo L, Succu S, et al (2007) Oocyte cryopreservation: oocyte assessment and strategies for improving survival. Reprod Fertil Dev 19:13–23. doi: 10.1071/rd06126
Liebermann J (2002) Potential Importance of Vitrification in Reproductive Medicine. Biology of reproduction 67:1671–1680. doi: 10.1095/biolreprod.102.006833
Liu C, Su K, Shang W, et al (2020) Higher implantation and live birth rates with laser zona pellucida breaching than thinning in single frozen-thawed blastocyst transfer. Lasers Med Sci. doi: 10.1007/s10103-019-02946-7
Lu X, Liu Y, Cao X, et al (2019) Laser-assisted hatching and clinical outcomes in frozen-thawed cleavage-embryo transfers of patients with previous repeated failure. Lasers Med Sci 34:1137–1145. doi: 10.1007/s10103-018-02702-3
Men H, Agca Y, Riley LK, Critser JK (2006) Improved survival of vitrified porcine embryos after partial delipation through chemically stimulated lipolysis and inhibition of apoptosis. Theriogenology 66:2008–2016. doi: 10.1016/j.theriogenology.2006.05.018
Moawad M, Hussein HA, Abd El-Ghani M, et al (2019) Effects of cryoprotectants and cryoprotectant combinations on viability and maturation rates of Camelus dromedarius oocytes vitrified at germinal vesicle stage. Reprod Domest Anim 54:108–117. doi: 10.1111/rda.13319
Mostagir A, Elmetwally M, Montaser A, Zaabel S (2019) Effects of L Carnitine and Cryoprotectants on Viability Rate of Immature Buffalo Oocytes in vitro After Vitrification. AJVS 62:45. doi: 10.5455/ajvs.57457
Nagashima H, Kashiwazaki N, Ashman RJ, et al (1995) Cryopreservation of porcine embryos. Nature 374:416. doi: 10.1038/374416a0
Peng J, Li Q, Wigglesworth K, et al (2013) Growth differentiation factor 9:bone morphogenetic protein 15 heterodimers are potent regulators of ovarian functions. Proc Natl Acad Sci USA 110:E776-85. doi: 10.1073/pnas.1218020110
Phongnimitr T, Liang Y, Srirattana K, et al (2013) Effect of L-carnitine on maturation, cryo-tolerance and embryo developmental competence of bovine oocytes. Anim Sci J 84:719–725. doi: 10.1111/asj.12067
Räty M, Ketoja E, Pitkänen T, et al (2011) In vitro maturation supplements affect developmental competence of bovine cumulus-oocyte complexes and embryo quality after vitrification. Cryobiology 63:245–255. doi: 10.1016/j.cryobiol.2011.09.134
Romek M, Gajda B, Krzysztofowicz E, Smorag Z (2009) Lipid content of non-cultured and cultured pig embryo. Reprod Domest Anim 44:24–32. doi: 10.1111/j.1439-0531.2007.00984.x
Ruppert-Lingham CJ, Paynter SJ, Godfrey J, et al (2003) Developmental potential of murine germinal vesicle stage cumulus-oocyte complexes following exposure to dimethylsulphoxide or cryopreservation: loss of membrane integrity of cumulus cells after thawing. Hum Reprod 18:392–398. doi: 10.1093/humrep/deg071
Sadeesh EM, Sikka P, Balhara AK, Balhara S (2016) Developmental competence and expression profile of genes in buffalo (Bubalus bubalis) oocytes and embryos collected under different environmental stress. Cytotechnology 68:2271–2285. doi: 10.1007/s10616-016-0022-y
Schiewe MC, Zozula S, Nugent N, et al (2017) Modified microsecure vitrification: A safe, simple and highly effective cryopreservation procedure for human blastocysts. J Vis Exp. doi: 10.3791/54871
Scholz EC, Navas CG (2014) The Maining of Cryopreservation for in-vitro Fertilization Patients. Recent advances in cryopreservation. doi: 10.5772/58746
Sharma GT, Dubey PK, Chandra V (2010) Morphological changes, DNA damage and developmental competence of in vitro matured, vitrified-thawed buffalo (Bubalus bubalis) oocytes: A comparative study of two cryoprotectants and two cryodevices. Cryobiology 60:315–321. doi: 10.1016/j.cryobiol.2010.02.006
Son W-Y, Henderson S, Cohen Y, et al (2019) Immature oocyte for fertility preservation. Front Endocrinol (Lausanne) 10:464. doi: 10.3389/fendo.2019.00464
Sprícigo JF, Morató R, Arcarons N, et al (2017) Assessment of the effect of adding L-carnitine and/or resveratrol to maturation medium before vitrification on in vitro-matured calf oocytes. Theriogenology 89:47–57. doi: 10.1016/j.theriogenology.2016.09.035
Sutton-McDowall ML, Feil D, Robker RL, et al (2012) Utilization of endogenous fatty acid stores for energy production in bovine preimplantation embryos. Theriogenology 77:1632–1641. doi: 10.1016/j.theriogenology.2011.12.008
Vajta G, Holm P, Kuwayama M, et al (1998a) Open pulled straw (OPS) vitrification: A new way to reduce cryoinjuries of bovine ova and embryos. Mol Reprod Dev 51:53–58. doi: 10.1002/(SICI)1098-2795(199809)51:1<53::AID-MRD6>3.0.CO;2-V
Vajta G, Holm P, Kuwayama M, et al (1998b) Open pulled straw (OPS) vitrification: A new way to reduce cryoinjuries of bovine ova and embryos. Mol Reprod Dev 51:53–58. doi: 10.1002/(SICI)1098-2795(199809)51:1<53::AID-MRD6>3.0.CO;2-V
Vanella A, Russo A, Acquaviva R, et al (2000) L -propionyl-carnitine as superoxide scavenger, antioxidant, and DNA cleavage protector. Cell Biol Toxicol 16:99–104. doi: 10.1023/a:1007638025856
Wani NA, Misra AK, Maurya SN (2004) Maturation rates of vitrified-thawed immature buffalo (Bubalus bubalis) oocytes: effect of different types of cryoprotectants. Anim Reprod Sci 84:327–335. doi: 10.1016/j.anireprosci.2004.02.007
Wassarman PM (2008) Zona pellucida glycoproteins. J Biol Chem 283:24285–24289. doi: 10.1074/jbc.R800027200
Yamada C, Caetano HVA, Simões R, et al (2007) Immature bovine oocyte cryopreservation: comparison of different associations with ethylene glycol, glycerol and dimethylsulfoxide. Anim Reprod Sci 99:384–388. doi: 10.1016/j.anireprosci.2006.07.001
Yeo CX, Gilchrist RB, Thompson JG, Lane M (2008) Exogenous growth differentiation factor 9 in oocyte maturation media enhances subsequent embryo development and fetal viability in mice. Hum Reprod 23:67–73. doi: 10.1093/humrep/dem140
Zare Z, Abouhamzeh B, Masteri Farahani R, et al (2017) Supplementation of L-carnitine during in vitro maturation of mouse oocytes affects expression of genes involved in oocyte and embryo competence: An experimental study. Int J Reprod Biomed (Yazd) 15:779–786.
Zhao X-M, Du W-H, Wang D, et al (2011a) Recovery of mitochondrial function and endogenous antioxidant systems in vitrified bovine oocytes during extended in vitro culture. Mol Reprod Dev 78:942–950. doi: 10.1002/mrd.21389
Zhao X-M, Du W-H, Wang D, et al (2011b) Effect of cyclosporine pretreatment on mitochondrial function in vitrified bovine mature oocytes. Fertil Steril 95:2786–2788. doi: 10.1016/j.fertnstert.2011.04.089
Zhou C, Zhou G-B, Zhu S-E, et al (2007) Open-pulled straw (OPS) vitrification of mouse hatched blastocysts. Anim Biotechnol 18:45–54. doi: 10.1080/10495390600826412
Zolini AM, Carrascal-Triana E, Ruiz de King A, et al (2019) Effect of addition of l-carnitine to media for oocyte maturation and embryo culture on development and cryotolerance of bovine embryos produced in vitro. Theriogenology 133:135–143. doi: 10.1016/j.theriogenology.2019.05.005