Animal Reproduction (AR)
Animal Reproduction (AR)

In vitro embryo production in small ruminants: what is still missing?

Joanna Maria Gonçalves Souza-Fabjan; Gabriela Ramos Leal; Clara Ana Santos Monteiro; Ribrio Ivan Tavares Pereira Batista; Nathalia Oliveira Barbosa; Vicente José Figueirêdo Freitas

Downloads: 0
Views: 33


In vitro embryo production (IVEP) is an extremely important tool for genetic improvement in livestock and it is the biotechnology that has grown the most recently. However, multiple ovulation followed by embryo transfer is still considered the leading biotechnology for embryo production in small ruminants. This review aimed to identify what is still missing for more efficient diffusion of IVEP in small ruminants, going through the IVEP steps and highlighting the main factors affecting the outcomes. Oocyte quality is essential for the success of IVEP and an aspect to be considered in small ruminants is their reproductive seasonality and strategies to mitigate the effect of season. The logistics for oocyte collection from live females is more complex than in cattle, and tools to simplify this collection system and/or to promote an alternative way of recovering oocytes may be an important point in this scenario. The heterogeneity of oocytes collected from growing follicles in live females or from ovaries collected from abattoirs remains a challenge, and there is a demand to standardize/homogenize the hormonal stimulatory protocols and IVM protocols for each source of oocytes. The use of sexed semen is technically possible, however the low market demand associated with the high costs of the sexing process prevents the routine use of this technique, but its higher availability is an important aspect aiming for greater dissemination of IVEP. New noninvasive approaches for embryo selection are key factors since the selection for transfer or cryopreservation is another difficulty faced among laboratories. Embryo selection is based on morphological traits, although these are not necessarily reliable in predicting pregnancy. Several issues described in this review must be considered by researchers in other to promote the diffusion of IVEP in small ruminants.


goat, IVM, IVC, sheep


Alberio R, Olivera J, Roche A, Alabart J, Folch J. Performance of a modified ovum pick-up system using three different FSH stimulation protocols in ewes. Small Rumin Res. 2002;46(2-3):81-7.

Albuz FK, Sasseville M, Lane M, Armstrong DT, Thompson JG, Gilchrist RB. Simulated physiological oocyte maturation (SPOM): a novel in vitro maturation system that substantially improves embryos yield and pregnancy outcomes. Hum Reprod. 2010;25(12):2999-3011. PMid:20870682.

Almeida KC, Pereira AF, Alcantara AS No, Avelar SR, Bertolini L, Bertolini M, Freitas VJ, Melo LM. Real-time qRT-PCR analysis of EGF receptor in cumulus-oocyte complexes recovered by laparoscopy in hormonally treated goats. Zygote. 2011;19(2):127-36. PMid:20663235.

An L, Liu J, Du Y, Liu Z, Zhang F, Liu Y, Zhu X, Ling P, Chang S, Hu Y, Li Y, Xu B, Yang L, Xue F, Presicce GA, Du F. Synergistic effect of cysteamine, leukemia inhibitory factor, and Y27632 on goat oocyte maturation and embryo development in vitro. Theriogenology. 2018;108:56-62. PMid:29197293.

Anzalone DA, Palazzese L, Czernik M, Sabatucci A, Valbonetti L, Capra E, Loi P. Controlled spermatozoa-oocyte interaction improves embryo quality in sheep. Sci Rep. 2021;11(1):22629. PMid:34799642.

Ariu F, Bogliolo L, Pinna A, Malfatti L, Innocenzi P, Falchi L, Bebbere D, Ledda S. Cerium oxide nanoparticles (CeO2 NPs) improve the developmental competence of in vitro-matured prepubertal ovine oocytes. Reprod Fertil Dev. 2017;29(5):1046-56. PMid:28442051.

Armstrong DT, Irvine BJ, Earl CR, McLean D, Seamark RF. Gonadotropin stimulation regimens for follicular aspiration and in vitro embryo production from calf oocytes. Theriogenology. 1994;42(7):1227-36. PMid:16727627.

Avelar SRG, Moura RR, Sousa FC, Pereira AF, Almeida KC, Melo CHS, Teles-Filho ACA, Baril G, Melo LM, Teixeira DIA, Freitas VJF. Oocyte production and in vitro maturation in Canindé goats following hormonal ovarian stimulation. Anim Reprod. 2012;9(1):27-32.

Bai J, Hou J, Guan H, Yan F, Cui X, Liu L, Wang S, An X. Effect of 2-mercaptoethanol and cysteine supplementation during in vitro maturation on the developmental competence of oocytes from hormone-stimulated lambs. Theriogenology. 2008;70(5):758-64. PMid:18603288.

Baldassarre H, Furnus CC, Matos DG, Pessi H. In vitro production of sheep embryos using laparoscopic folliculocentesis: alternative gonadotrophin treatments for stimulation of oocyte donors. Theriogenology. 1996;45(3):707-17. PMid:16727833.

Baldassarre H, Rao KM, Neveu N, Brochu E, Begin I, Behboodi E, Hockley DK. Laparoscopic ovum pick-up followed by in vitro embryo production for the reproductive rescue of aged goats of high genetic value. Reprod Fertil Dev. 2007;19(5):612-6. PMid:17601408.

Baldassarre H, Wang B, Kafidi N, Keefer C, Lazaris A, Karatzas CN. Advances in the production and propagation of transgenic goats using laparoscopic ovum pick-up and in vitro embryo production technologies. Theriogenology. 2002;57(1):275-84. PMid:11775975.

Baldassarre H. Practical aspects for implementing in vitro embryo production and cloning programs in sheep and goats. Anim Reprod. 2012;9:188-94.

Bathgate R, Mace N, Heasman K, Evans G, Maxwell WM, de Graaf SP. Birth of kids after artificial insemination with sex-sorted, frozen-thawed goat spermatozoa. Reprod Domest Anim. 2013;48(6):893-8. PMid:23701168.

Bavister BD. Culture of preimplantation embryos: facts and artifacts. Hum Reprod Update. 1995;1(2):91-148. PMid:15726768.

Bebbere D, Nieddu SM, Ariu F, Piras D, Ledda S. 3D liquid marble microbioreactors support in vitro maturation of prepubertal ovine oocytes and affect expression of oocyte-specific factors. Biology. 2021;10(11):1101. PMid:34827093.

Beilby KH, de Graaf SP, Evans G, Maxwell WMC, Wilkening S, Wrenzycki C, Grupen CG. Quantitative mRNA expression in ovine blastocysts produced from X- and Y-chromosome bearing sperm, both in vitro and in vivo. Theriogenology. 2011;76(3):471-81. PMid:21497386.

Berlinguer F, Gonzalez-Bulnes A, Succu S, Leoni G, Mossa F, Bebbere D, Ariznavarreta C, Tresguerres JA, Veiga-Lopez A, Naitana S. Effects of progestagens on follicular growth and oocyte developmental competence in FSH-treated ewes. Domest Anim Endocrinol. 2007;32(4):303-14. PMid:16713166.

Bormann CL, Ongeri EM, Krisher RL. The effect of vitamins during maturation of caprine oocytes on subsequent developmental potential in vitro. Theriogenology. 2003;59(5-6):1373-80. PMid:12527083.

Bragança GM, Alcântara-Neto AS, Batista RITP, Brandão FZ, Freitas VJF, Mermillod P, Souza-Fabjan JMG. Oviduct fluid during IVF moderately modulates polyspermy in in vitro-produced goat embryos during the non-breeding season. Theriogenology. 2021a;168:59-65. PMid:33857909.

Bragança GM, Batista RIT, Souza-Fabjan JMG, Alfradique VAP, Arashiro EKN, Pinto PHN, Santos JDR, Camargo LSA, Menchaca A, Fonseca JF, Brandão FZ. Exogenous progestogens differentially alter gene expression of immature cumulus-oocyte complexes in sheep. Domest Anim Endocrinol. 2021b;74:106518. PMid:32711283.

Bragança GM, Batista RITP, Souza-Fabjan JMG, Alfradique VAP, Arashiro EKN, Cosentino IO, Pinto PHN, Camargo LSA, Fonseca JF, Brandão FZ. Dose and administration protocol for FSH used for ovarian stimulation affect gene expression in sheep cumulus-oocyte complexes. Reprod Fertil Dev. 2018;30(9):1234-44. PMid:29579410.

Brown BW, Radziewic T. Production of sheep embryos in vitro and development of progeny following single and twin embryo transfers. Theriogenology. 1998;49(8):1525-36. PMid:10732016.

Byrne GP, Lonergan P, Wade M, Duffy P, Donovan A, Hanrahan JP, Boland MP. Effect of freezing rate of ram spermatozoa on subsequent fertility in vivo and in vitro. Anim Reprod Sci. 2000;62(4):265-75. PMid:10924829.

Cannon MJ, Menino AR Jr. Changes in the bovine zona pellucida induced by plasmin or embryonic plasminogen activator. Mol Reprod Dev. 1998;51(3):330-8.<330::AID-MRD13>3.0.CO;2-6. PMid:9771654.

Castro T, Rubianes E, Menchaca A, Rivero A. Ovarian dynamics, serum estradiol and progesterone concentrations during the interovulatory interval in goats. Theriogenology. 1999;52(3):399-411. PMid:10734375.

Catalá MG, Izquierdo D, Rodríguez-Prado M, Hammami S, Paramio MT. Effect of oocyte quality on blastocyst development after in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in a sheep model. Fertil Steril. 2012;97(4):1004-8. PMid:22265000.

Catalá MG, Roura M, Izquierdo D, Morato R, Hammami S, Paramio MT. Blastocyst development, MPF activity and ATP content of lamb oocytes supplemented with insulin-transferrin-selenium (ITS) and ascorbic acid at IVM. Small Rumin Res. 2013;112(1-3):103-7.

Catalá MG, Roura M, Soto-Heras S, Menéndez I, Contreras-Solis I, Paramio MT, Izquierdo D. Effect of season on intrafollicular fatty acid concentrations and embryo production after in vitro fertilization and parthenogenic activation of prepubertal goat oocytes. Small Rumin Res. 2018;168:82-6.

Chian R, Okuda K, Niwa K. Influence of cumulus cells on in vitro fertilization of bovine oocytes derived from in vitro maturation. Anim Reprod Sci. 1995;38(1-2):37-48.

Cocero MJ, Marigorta P, Novillo F, Folch J, Sánchez P, Alabart JL, Lahoz B. Ovine oocytes display a similar germinal vesicle configuration and global DNA methylation at prepubertal and adult ages. Theriogenology. 2019;138:154-63. c10.1016/j.theriogenology.2019.07.011. PMid:31357118.

Cognié Y, Baril G, Poulin N, Mermillod P. Current status of embryo technologies in sheep and goat. Theriogenology. 2003;59(1):171-88. PMid:12499027.

Cognié Y, Poulin N, Locatelli Y, Mermillod P. State-of-the-art production, conservation and transfer of in-vitro-produced embryos in small ruminants. Reprod Fertil Dev. 2004;16(4):437-45. PMid:15315742.

Cognié Y, Poulin N. Developmental competence of goat oocytes is increased after in vitro maturation with follicular fluid from goats stimulated by gonadotropins. In: Proceedings of 14th International Congress on Animal Reproduction; 2000 Jul 2-6; Stockholm, Sweden. Stockholm: ICAR; 2000. p. 39.

Colas C, James P, Howes L, Jones R, Cebrian-Perez JA, Muiño-Blanco T. Cyclic-AMP initiates protein tyrosine phosphorylation independent of cholesterol efflux during ram sperm capacitation. Reprod Fertil Dev. 2008;20(6):649-58. PMid:18671912.

Conceição JC, Moura MT, Ferreira-Silva JC, Ramos-Deus P, Silva PG, Cantanhêde LF, Chaves RM, Lima PF, Oliveira MA. Use of retinoids during oocyte maturation diminishes apoptosis in caprine embryos. Acta Vet Hung. 2015;63(2):234-42. PMid:26051262.

Cox JF, Alfaro V. In vitro fertilization and development of OPU derived goat and sheep oocytes. Reprod Domest Anim. 2007;42(1):83-7. PMid:17214779.

Crozet N, Ahmed-Ali M, Dubos MP. Developmental competence of goat oocytes from follicles of different size categories following maturation, fertilization and culture in vitro. J Reprod Fertil. 1995;103(2):293-8. PMid:7616502.

De AK, Malakar D, Akshey YS, Jena MK, Garg S, Dutta R, Sahu S. In vitro development of goat (Capra hircus) embryos following cysteamine supplementation of the in vitro maturation and in vitro culture media. Small Rumin Res. 2011;96(2-3):185-90.

de Graaf SP, Evans G, Gillan L, Guerra MM, Maxwell WM, O’Brien JK. The influence of antioxidant, cholesterol and seminal plasma on the in vitro quality of sorted and non-sorted ram spermatozoa. Theriogenology. 2007;67(2):217-27. PMid:16934321.

de Lamirande E, Gagnon C. Impact of reactive oxygen species on spermatozoa: a balancing act between beneficial and detrimental effects. Hum Reprod. 1995;10(Suppl 1):15-21. PMid:8592032.

De Smedt V, Crozet N, Ahmed-Ali M, Martino A, Cognié Y. In vitro maturation and fertilization of goat oocytes. Theriogenology. 1992;37(5):1049-60. PMid:16727103.

Dunglison GF, Jane SD, McCaul TF, Chad JE, Fleming TP, Kaye PL. Stimulation of endocytosis in mouse blastocysts by insulin: a quantitative morphological analysis. J Reprod Fertil. 1995;105(1):115-23. PMid:7490702.

Dunglison GF, Kaye PL. Insulin regulates protein metabolism in mouse blastocysts. Mol Reprod Dev. 1993;36(1):42-8. PMid:8398129.

Evans AC, Duffy P, Hynes N, Boland MP. Waves of follicle development during the estrous cycle in sheep. Theriogenology. 2000;53(3):699-715. PMid:10735037.

Falchi L, Ledda S, Zedda MT. Embryo biotechnologies in sheep: achievements and new improvements. Reprod Domest Anim. 2022;57(S5):22-33. PMid:35437835.

Fathi M, Elkarmoty AF. Effect of adding growth factors during in vitro maturation on the developmental potentials of ewe oocytes selected by brilliant cresyl blue staining. Vet World. 2021;14(2):452-6. PMid:33776311.

Ferreira EM, Vireque AA, Adona PR, Meirelles FV, Ferriani RA, Navarro PA. Cytoplasmic maturation of bovine oocytes: structural and biochemical modifications and acquisition of developmental competence. Theriogenology. 2009;71(5):836-48. PMid:19121865.

Ferreira-Silva JC, Silva RLO, Vieira JIT, Silva JB, Tavares LS, Silva FAC, Pena EPN, Chaves MS, Moura MT, Junior TC, Benko-Iseppon AM, Freitas VJF, Oliveira MAL. Evaluation of quality and gene expression of goat embryos produced in vivo and in vitro after cryopreservation. Cryobiology. 2021;101:115-24. PMid:33964298.

Fieni F, Pellerin JL, Roux C, Poulin N, Baril G, Fatet A, Valas S, Chatagnon G, Mermillod P, Guignot F. Can caprine arthritis encephalitis virus (CAEV) be transmitted by in vitro fertilization with experimentally infected sperm? Theriogenology. 2012;77(3):644-51. PMid:22015154.

Fonseca JF, Oliveira MEF, Brandão FZ, Batista RITP, Garcia AR, Bartlewski PM, Souza-Fabjan JMG. Non-surgical embryo transfer in goats and sheep: the Brazilian experience. Reprod Fertil Dev. 2018;31(1):17-26. PMid:32188539.

Fryc K, Nowak A, Kij B, Kochan J, Bartlewski PM, Murawski M. Timing of cleavage divisions determined with time-lapse imaging is linked to blastocyst formation rates and quality of in vitro-produced ovine embryos. Theriogenology. 2021;159:147-52. PMid:33157452.

Galli C, Crotti G, Notari C, Turini P, Duchi R, Lazzari G. Embryo production by ovum pick up from live donors. Theriogenology. 2001;55(6):1341-57. PMid:11327688.

Gandolfi F, Moor RM. Stimulation of early embryonic development in the sheep by co-culture with oviduct epithelial cells. J Reprod Fertil. 1987;81(1):23-8. PMid:3668954.

Garcia-Garcia RM, Ward F, Fair S, O’Meara CM, Wade M, Duffy P, Lonergan P. Development and quality of sheep embryos cultured in commercial G1.3/G2.3 sequential media. Anim Reprod Sci. 2007;98(3-4):233-40. PMid:16621355.

Gardner DK, Lane M. Culture and selection of viable blastocysts: a feasible proposition for human IVF? Hum Reprod Update. 1997;3(4):367-82. PMid:9459282.

Ghaffarilaleh V, Fouladi-Nashta A, Paramio MT. Effect of α-linolenic acid on oocyte maturation and embryo development of prepubertal sheep oocytes. Theriogenology. 2014;82(5):686-96. PMid:25015785.

Gibbons A, Bonnet FP, Cueto MI, Catala M, Salamone DF, Gonzalez-Bulnes A. Procedure for maximizing oocyte harvest for in vitro embryo production in small ruminants. Reprod Domest Anim. 2007;42(4):423-6. PMid:17635781.

Gilchrist RB, Thompson JG. Oocyte maturation: emerging concepts and technologies to improve developmental potential in vitro. Theriogenology. 2007;67(1):6-15. PMid:17092551.

Gómez MC, Catt JW, Evans G, Maxwell WM. Cleavage, development and competence of sheep embryos fertilized by intracytoplasmic sperm injection and in vitro fertilization. Theriogenology. 1998;49(6):1143-54. PMid:10732052.

González R, Ruiz-León Y, Gomendio M, Roldan ERS. The effect of glucocorticoids on ERK-1/2 phosphorylation during maturation of lamb oocytes and their subsequent fertilization and cleavage ability in vitro. Reprod Toxicol. 2010;29(2):198-205. PMid:19892010.

Grasa P, Cebrián-Pérez JA, Muiño-Blanco T. Signal transduction mechanisms involved in in vitro ram sperm capacitation. Reproduction. 2006;132(5):721-32. PMid:17071773.

Greyling JPC, van der Nest M. Synchronization of oestrus in goats: dose effect of progestagen. Small Rumin Res. 2000;36(2):201-7. PMid:10760457.

Guler A, Poulin N, Mermillod P, Terqui M, Cognié Y. Effect of growth factors, EGF and IGF-I, and estradiol on in vitro maturation of sheep oocytes. Theriogenology. 2000;54(2):209-18. PMid:11003302.

Hagemann LJ, Weilert LL, Beaumont SE, Tervit HR. Development of bovine embryos in single in vitro production (sIVP) systems. Mol Reprod Dev. 1998;51(2):143-7.<143::AID-MRD3>3.0.CO;2-Q. PMid:9740321.

Hammami S, Morató R, Romaguera R, Roura M, Catalá MG, Paramio MT, Mogas T, Izquierdo D. Developmental competence and embryo quality of small oocytes from pre-pubertal goats cultured in IVM medium supplemented with low level of hormones, insulin-transferrin-selenium and ascorbic acid. Reprod Domest Anim. 2013;48(2):339-44. PMid:22908901.

Han D, Zhao BT, Liu Y, Li JJ, Wu YG, Lan GC, Tan JH. Interactive effects of low temperature and roscovitine (ROS) on meiotic resumption and developmental potential of goat oocytes. Mol Reprod Dev. 2008;75(5):838-46. PMid:17926351.

Han YM, Abeydeera LR, Kim JH, Moon HB, Cabot RA, Day BN, Prather RS. Growth retardation of inner cell mass cells in polyspermic porcine embryos produced in vitro. Biol Reprod. 1999;60(5):1110-3. http://dx.doi.org10.1095/biolreprod60.5.1110. PMid:10208971.

Herrick JR, Behboodi E, Memili E, Blash S, Echelard Y, Krisher RL. Effect of macromolecule supplementation during in vitro maturation of goat oocytes on developmental potential. Mol Reprod Dev. 2004;69(3):338-46. PMid:15349846.

Hollinshead FK, Evans G, Evans KM, Catt SL, Maxwell WM, O’Brien JK. Birth of lambs of a pre-determined sex after in vitro production of embryos using frozen-thawed sex-sorted and re-frozen-thawed ram spermatozoa. Reproduction. 2004;127(5):557-68. PMid:15129011.

Huang M, Cao XY, He QF, Yang HW, Chen YZ, Zhao JL, Ma HW, Kang J, Liu J, Quang FS. Alkaline semen diluent combined with R848 for separation and enrichment of dairy goat X-sperm. J Dairy Sci. 2022;105(12):10020-32. PMid:36270871.

Izquierdo D, Villamediana P, Palomo MJ, Mogas T, Paramio MT. Effect of sperm capacitation and fertilization media on IVF and early embryo development of prepubertal goat oocytes. Theriogenology. 1998;49(8):1501-13. PMid:10732014.

Kątska-Książkiewicz L, Opiela J, Ryńska B. Effects of oocyte quality, semen donor and embryo co-culture system on the efficiency of blastocyst production in goats. Theriogenology. 2007;68(5):736-44. PMid:17651793.

Kelly JM, Kleemann DO, Walker SK. Enhanced efficiency in the production of offspring from 4 to 8-week-old lambs. Theriogenology. 2005;63(7):1876-90. PMid:15823345.

Keskintepe L, Darwish GM, Younis AI, Brackett BG. In vitro development of morulae from immature caprine oocytes. Zygote. 1994;2(2):97-102. PMid:7874460.

Keskintepe L, Simplicio AA, Brackett BG. Caprine blastocyst development after in vitro fertilization with spermatozoa frozen in different extenders. Theriogenology. 1998;49(7):1265-74. PMid:10732064.

Koeman J, Keefer CL, Baldassarre H, Downey BR. Developmental competence of prepubertal and adult goat oocytes cultured in semi-defined media following laparoscopic recovery. Theriogenology. 2003;60(5):879-89. PMid:12935865.

Kouamo J, Kharche SD. A comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured caprine oocytes. Iran J Vet Res. 2015;16(1):20-4. PMid:27175145.

Krisher RL, Schoolcraft WB, Katz-Jaffe MG. Omics as a window to view embryo viability. Fertil Steril. 2015;103(2):333-41. PMid:25639968.

Le Gal F, Gall L, De Smedt V. Changes in protein synthesis pattern during in vitro maturation of goat oocytes. Mol Reprod Dev. 1992;32(1):1-8. PMid:1515144.

Leal GR, Monteiro CAS, Carvalheira LR, Souza-Fabjan JMG. The Simulated Physiological Oocyte Maturation (SPOM) system in domestic animals: a systematic review. Theriogenology. 2022;188:90-9. PMid:35688043.

Leal GR, Monteiro CAS, Souza-Fabjan JMG, Vasconcelos COP, Nogueira LAG, Ferreira AMR, Serapião RV. Role of cAMP modulator supplementations during oocyte in vitro maturation in domestic animals. Anim Reprod Sci. 2018;199:1-14. PMid:30449707.

Ledda S, Bogliolo L, Calvia P, Leoni G, Naitana S. Meiotic progression and developmental competence of oocytes collected from juvenile and adult ewes. J Reprod Fertil. 1997;109(1):73-8. PMid:9068416.

Ledda S, Bogliolo L, Leoni G, Naitana S. Production and lambing rate of blastocysts derived from in vitro matured oocytes after gonadotropin treatment of prepubertal ewes. J Anim Sci. 1999;77(8):2234-9. PMid:10462004.

Leoni GG, Palmerini MG, Satta V, Succu S, Pasciu V, Zinellu A, Carru C, Macchiarelli G, Nottola SA, Naitana S, Berlinguer F. Differences in the kinetic of the first meiotic division and in active mitochondrial distribution between prepubertal and adult oocytes mirror differences in their developmental competence in a sheep model. PLoS One. 2015;10(4):e0124911. PMid:25893245.

Leoni GG, Rosati I, Succu S, Bogliolo L, Bebbere D, Berlinguer F, Ledda S, Naitana S. A low oxygen atmosphere during IVF accelerates the kinetic of formation of in vitro produced ovine blastocysts. Reprod Domest Anim. 2007;42(3):299-304. PMid:17506809.

Locatelli Y, Poulin N, Baril G, Touzé JL, Fatet A, Beckers JF, Mermillod P. 207 in vitro maturation treatment affects developmental competence of laparoscopic ovum pickup-derived oocytes in follicle- stimulating hormone-stimulated goats. Reprod Fertil Dev. 2008;20:182-3.

Lorenzo-Torres A, Rangel-Santos R, Ruíz-Flores A, Ambríz-García DA. In vitro embryo production from ewes at different physiological stages. J Vet Sci. 2023;24(1):e10. PMid:36560839.

Majeed A, Omar A, Alrawi S, Ahmed K. Effect of collection methods on oocyte recovery rate in sheep. Res J Biotechnol. 2019;14:262-4.

Mara L, Sanna D, Casu S, Dattena M, Muñoz IM. Blastocyst rate of in vitro embryo production in sheep is affected by season. Zygote. 2014;22(3):366-71. PMid:23458093.

Masala L, Ariu F, Bogliolo L, Bellu E, Ledda S, Bebbere D. Delay in maternal transcript degradation in ovine embryos derived from low competence oocytes. Mol Reprod Dev. 2018;85(5):427-39. PMid:29542856.

Mastromonaco GF, Gonzalez-Grajales AL. Reproduction in female wild cattle: influence of seasonality on ARTs. Theriogenology. 2020;150:396-404. PMid:32081408.

McLennan HJ, Saini A, Dunning KR, Thompson JG. Oocyte and embryo evaluation by AI and multi-spectral auto-fluorescence imaging: livestock embryology needs to catch-up to clinical practice. Theriogenology. 2020;150:255-62. PMid:32088032.

Meirelles FV, Caetano AR, Watanabe YF, Ripamonte P, Carambula SF, Merighe GK, Garcia SM. Genome activation and developmental block in bovine embryos. Anim Reprod Sci. 2004;82-83:13-20. PMid:15271440.

Melo-Sterza AF, Poehland R. Lipid metabolism in bovine oocytes and early embryos under in vivo, in vitro, and stress conditions. Int J Mol Sci. 2021;22(7):3421. PMid:33810351.

Menchaca A, Cuadro F, Santos-Neto PC, Bosolasco D, Barrera N, de Brun V, Crispo M. Oocyte developmental competence is improved by relatively greater circulating progesterone concentrations during preovulatory follicular growth. Anim Reprod Sci. 2018;195:321-8. PMid:31262405.

Menéndez-Blanco I, Soto-Heras S, Catalá MG, Roura M, Izquierdo D, Paramio MT. Effect of crocetin added to IVM medium for prepubertal goat oocytes on blastocyst outcomes after IVF, intracytoplasmic sperm injection and parthenogenetic activation. Theriogenology. 2020;155:70-6. PMid:32623132.

Mermillod P, Dalbiès-Tran R, Uzbekova S, Thélie A, Traverso JM, Perreau C, Papillier P, Monget P. Factors affecting oocyte quality: who is driving the follicle? Reprod Domest Anim. 2008;43(S2):393-400. PMid:18638152.

Modina S, Leoni GG, Lodde V, Naitana S, Pirani S, Succu S, Berlinguer F, Luciano AM. Involvement of E-cadherin in early in vitro development of adult and juvenile sheep embryos. Reprod Fertil Dev. 2010;22(2):468-77. PMid:20047732.

Mossa F, Leoni GG, Berlinguer F, Succu S, Madeddu M, Bebbere D, Naitana S. Recovery of COCs from ovaries with high follicle numbers enhances in vitro embryo yield in sheep. Anim Reprod Sci. 2008;109(1-4):134-45. PMid:18162333.

Nõmm M, Porosk R, Pärn P, Kilk K, Soomets U, Kõks S, Jaakma Ü. In vitro culture and non-invasive metabolic profiling of single bovine embryos. Reprod Fertil Dev. 2019;31(2):306-14. PMid:30092912.

O’Brien JK, Catt SL, Ireland KA, Maxwell WM, Evans G. In vitro and in vivo developmental capacity of oocytes from prepubertal and adult sheep. Theriogenology. 1997;47(7):1433-43. PMid:16728089.

Paramio MT, Izquierdo D. Recent advances in in vitro embryo production in small ruminants. Theriogenology. 2016;86(1):152-9. PMid:27157391.

Paramio M-T, Izquierdo P. Current status of in vitro embryo production in sheep and goats. Reprod Domest Anim. 2014;49(S4):37-48. PMid:25277431.

Parrilla I, Vazquez JM, Roca J, Martinez EA. Flow cytometry identification of X- and Y-chromosome-bearing goat spermatozoa. Reprod Domest Anim. 2004;39(1):58-60. PMid:15129923.

Passos JRS, Guerreiro DD, Otávio KS, Santos-Neto PC, Souza-Neves M, Cuadro F, Nuñez-Olivera R, Crispo M, Bezerra MJB, Silva RF, Lima LF, Figueiredo JR, Bustamante-Filho IC, Menchaca A, Moura AA. Global proteomic analysis of preimplantational ovine embryos produced in vitro. Reprod Domest Anim. 2022;57(7):784-97. PMid:35377953.

Pawshe CH, Palanisamy A, Taneja M, Jain SK, Totey SM. Comparison of various maturation treatments on in vitro maturation of goat oocytes and their early embryonic development and cell numbers. Theriogenology. 1996;46(6):971-82. PMid:16727960.

Pierson J, Wang B, Neveu N, Sneek L, Côté F, Karatzas CN, Baldassarre H. Effects of repetition, interval between treatments and season on the results from laparoscopic ovum pick-up in goats. Reprod Fertil Dev. 2004;16(8):795-9. PMid:15740703.

Pinheiro JBS, Figueira LM, Correia LFL, Oliveira TA, Brair VL, Barros FFPC, Ascoli FO, Batista RITP, Brandão FZ, Souza-Fabjan JMG. The coasting time affects the quality of cumulus-oocyte complexes in superstimulated ewes. Theriogenology. 2023;196:236-43. PMid:36434845.

Piras AR, Menéndez-Blanco I, Soto-Heras S, Catalá MG, Izquierdo D, Bogliolo L, Paramio MT. Resveratrol supplementation during in vitro maturation improves embryo development of prepubertal goat oocytes selected by brilliant cresyl blue staining. J Reprod Dev. 2019;65(2):113-20. PMid:30606957.

Pradeep MA, Jagadeesh J, De AK, Kaushik JK, Malakar D, Kumar S, Dang AK, Das SK, Mohanty AK. Purification, sequence characterization and effect of goat oviduct-specific glycoprotein on in vitro embryo development. Theriogenology. 2011;75(6):1005-15. PMid:21196036.

Ptak G, Dattena M, Loi P, Tischner M, Cappai P. Ovum pick-up in sheep: efficiency of in vitro embryo production, vitrification and birth of offspring. Theriogenology. 1999a;52(6):1105-14. PMid:10735116.

Ptak G, Loi P, Dattena M, Tischner M, Cappai P. Offspring from one month old lambs: studies on the developmental capability of prepubertal oocytes. Biol Reprod. 1999b;61(6):1568-74. PMid:10570004.

Reader KL, Cox NR, Stanton JAL, Juengel JL. Mitochondria and vesicles differ between adult and prepubertal sheep oocytes during IVM. Reprod Fertil Dev. 2015;27(3):513-22. PMid:24456710.

Rho GJ, Hahnel AC, Betteridge KJ. Comparisons of oocyte maturation times and of three methods of sperm preparation for their effects on the production of goat embryos in vitro. Theriogenology. 2001;56(3):503-16. PMid:11516129.

Rizos D, Gutiérrez-Adán A, Pérez-Garnelo S, de la Fuente J, Boland MP, Lonergan P. Bovine embryo culture in the presence or absence of serum: implications for blastocyst development, cryotolerance, and messenger RNA expression. Biol Reprod. 2003;68(1):236-43. PMid:12493719.

Rizos D, Lonergan P, Boland MP, Arroyo-García R, Pintado B, de la Fuente J, Gutiérrez-Adán A. Analysis of differential messenger RNA expression between bovine blastocysts produced in different culture systems: implications for blastocyst quality. Biol Reprod. 2002;66(3):589-95. PMid:11870062.

Rodríguez C, Anel L, Alvarez M, Anel E, Boixo JC, Chamorro CA, Paz P. Ovum pick-up in sheep: a comparison between different aspiration devices for optimal oocyte retrieval. Reprod Domest Anim. 2006;41(2):106-13. PMid:16519714.

Rodríguez-Dorta N, Cognié Y, González F, Poulin N, Guignot F, Touzé JL, Baril G, Cabrera F, Alamo D, Batista M, Gracia A, Mermillod P. Effect of coculture with oviduct epithelial cells on viability after transfer of vitrified in vitro produced goat embryos. Theriogenology. 2007;68(6):908-13. PMid:17719625.

Rodríguez-González E, López-Béjar M, Velilla E, Paramio MT. Selection of prepubertal goat oocytes using the brilliant cresyl blue test. Theriogenology. 2002;57(5):1397-409. PMid:12054199.

Romaguera R, Moll X, Morató R, Roura M, Palomo MJ, Catalá MG, Jiménez-Macedo AR, Hammami S, Izquierdo D, Mogas T, Paramio MT. Prepubertal goat oocytes from large follicles result in similar blastocyst production and embryo ploidy than those from adult goats. Theriogenology. 2011;76(1):1-11. PMid:21295839.

Rose RD, Gilchrist RB, Kelly JM, Thompson JG, Sutton-McDowall ML. Regulation of sheep oocyte maturation using cAMP modulators. Theriogenology. 2013;79(1):142-8. PMid:23102843.

Saini S, Sharma V, Ansari S, Kumar A, Thakur A, Malik H, Kumar S, Malakar D. Folate supplementation during oocyte maturation positively impacts the folate-methionine metabolism in pre-implantation embryos. Theriogenology. 2022;182:63-70. PMid:35123312.

Samereh S, Hajarian H, Karamishabankareh H, Soltani L, Foroutanifar S. Effects of different concentrations of Chir98014 as an activator of Wnt/beta-catenin signaling pathway on oocyte in-vitro maturation and subsequent embryonic development in Sanjabi ewes. Reprod Domest Anim. 2021;56(7):965-71. PMid:33866629.

Sanchez DJD, Vasconcelos FR, Teles-Filho ACA, Viana AGA, Martins AMA, Sousa MV, Castro MS, Ricart CA, Fontes W, Bertolini M, Bustamante-Filho IC, Moura AA. Proteomic profile of pre-implantational ovine embryos produced in vivo. Reprod Domest Anim. 2021;56(4):586-603. PMid:33460477.

Sánchez-Ajofrín I, Peris-Frau P, García-Álvarez O, Fernández-Santos MDR, Montoro V, Garde JJ, Soler AJ. Serum supplementation during in vitro fertilization of sheep oocytes influences blastocyst quality through the differential abundance of mRNA transcripts. Reprod Domest Anim. 2022;57(S5):68-71. PMid:35575960.

Santos JD, Batista RI, Magalhães LC, Paula AR Jr, Souza SS, Salamone DF, Bhat MH, Teixeira DI, Freitas VJ, Melo LM. Overexpression of hyaluronan synthase 2 and gonadotropin receptors in cumulus cells of goats subjected to one-shot eCG/FSH hormonal treatment for ovarian stimulation. Anim Reprod Sci. 2016;170:15-24. PMid:27072623.

Santos-Neto PC, Vilariño M, Cuadro F, Barrera N, Crispo M, Menchaca A. Cumulus cells during in vitro fertilization and oocyte vitrification in sheep: remove, maintain or add? Cryobiology. 2020;92:161-7. PMid:31917962.

Sargison ND. The critical importance of planned small ruminant livestock health and production in addressing global challenges surrounding food production and poverty alleviation. N Z Vet J. 2020;68(3):136-44. PMid:31968203.

Shabankareh HK, Zandi M. Developmental potential of sheep oocytes cultured in different maturation media: effects of epidermal growth factor, insulin-like growth factor I, and cysteamine. Fertil Steril. 2010;94(1):335-40. PMid:19324348.

Shakir Hammoud S, Adil Jebur M. Effect of Epididymal Spermatozoa of Local Iraqi Goat on in vitro Fertilization and Evolution of Embryos. Arch Razi Inst. 2022;77(2):641-6. PMid:36284945.

Shi L, Yue W, Zhang J, Lv L, Ren Y, Yan P. Effect of ovarian cortex cells on nuclear maturation of sheep oocytes during in vitro maturation. Anim Reprod Sci. 2009;113(1-4):299-304. PMid:18790578.

Shirazi A, Motaghi E. The in vitro fertilization of ovine oocytes in the presence of oviductal cells and its effect on the expression of zygote arrest 1 (Zar1) and subsequent embryonic development. J Reprod Infertil. 2013;14(1):8-16. PMid:23926555.

Shirazi A, Shams-Esfandabadi N, Ahmadi E, Heidari B. Effects of growth hormone on nuclear maturation of ovine oocytes and subsequent embryo development. Reprod Domest Anim. 2010;45(3):530-6. PMid:19032427.

Sinclair KD, Maxfield EK, Robinson JJ, Maltin CA, McEvoy TG, Dunne LD, Young LE, Broadbent PJ. Culture of sheep zygotes can alter fetal growth and development. Theriogenology. 1997;47(1):380.

Sinclair KD, McEvoy TG, Maxfield EK, Maltin CA, Young LE, Wilmut I, Broadbent PJ, Robinson JJ. Aberrant fetal growth and development after in vitro culture of sheep zygotes. J Reprod Fertil. 1999;116(1):177-86. PMid:10505068.

Sirard MA, Richard F, Blondin P, Robert C. Contribution of the oocyte to embryo quality. Theriogenology. 2006;65(1):126-36. PMid:16256189.

Slimane W, Heyman Y, Lavergne Y, Humblot P, Renard JP. Assessing chromosomal abnormalities in two-cell bovine in vitro-fertilized embryos by using fluorescent in situ hybridization with three different cloned probes. Biol Reprod. 2000;62(3):628-35. PMid:10684804.

Snyder DA, Dukelow WR. Laparoscopic studies of ovulation, pregnancy diagnosis, and follicle aspiration in sheep. Theriogenology. 1974;2(6):143-8. PMid:4282076.

Sousa FC, Melo CHS, Teles ACA Fo, Avelar SRG, Moura AAA, Martins JAM, Freitas VJF, Teixeira DÍA. Ovarian follicular response to different hormonal stimulation treatments in Canindé goats. Anim Reprod Sci. 2011;125(1-4):88-93. PMid:21420805.

Souza JM, Duffard N, Bertoldo MJ, Locatelli Y, Corbin E, Fatet A, Freitas VJ, Mermillod P. Influence of heparin or the presence of cumulus cells during fertilization on the in vitro production of goat embryos. Anim Reprod Sci. 2013;138(1-2):82-9. PMid:23517856.

Souza-Fabjan JM, Locatelli Y, Duffard N, Corbin E, Batista RI, Freitas VJF, Beckers JF, Mermillod P. Intrinsic quality of goat oocytes already found denuded at collection for in vitro embryo production. Theriogenology. 2016;86(8):1989-98. PMid:27453560.

Souza-Fabjan JM, Locatelli Y, Duffard N, Corbin E, Touzé JL, Perreau C, Beckers JF, Freitas VJ, Mermillod P. In vitro embryo production in goats: slaughterhouse and laparoscopic ovum pick up-derived oocytes have different kinetics and requirements regarding maturation media. Theriogenology. 2014a;81(8):1021-31. PMid:24582267.

Souza-Fabjan JMG, Alves BRC, Batista RITP, Pereira AF, Melo LM, Freitas VJF, Oliveira MEF. Reproductive biotechnologies applied to the female sheep and goat. In: Yata VK, Mohanty AK, Lichtfouse E, editors. Sustainable agriculture reviews 59. Cham: Springer; 2023. p. 1-57.

Souza-Fabjan JMG, Batista RITP, Freitas VJF, Mermillod P. In vitro culture of embryos from LOPU-derived goat oocytes. Methods Mol Biol. 2019;2006:141-53. PMid:31230278.

Souza-Fabjan JMG, Correia LFL, Batista RITP, Locatelli Y, Freitas VJF, Mermillod P. Reproductive seasonality affects in vitro embryo production outcomes in adult goats. Animals. 2021;11(3):873. PMid:33803854.

Souza-Fabjan JMG, Locatelli Y, Freitas VJF, Mermillod P. Laparoscopic ovum pick up (LOPU) in goats: from hormonal treatment to oocyte possible destinations. R Bras Ci Vet. 2014b;21(1):3-11.

Souza-Fabjan JMG, Pereira AF, Melo CH, Sanchez DJ, Oba E, Mermillod P, Melo LM, Teixeira DI, Freitas VJ. Assessment of the reproductive parameters, laparoscopic oocyte recovery and the first embryos produced in vitro from endangered Canindé goats (Capra hircus). Reprod Biol. 2013;13(4):325-32. PMid:24287041.

Suresh A, Shukla MK, Kumar D, Shrivastava OP, Verma N. Simulated physiological oocyte maturation (SPOM) improves developmental competence of in vitro produced goat embryos. Theriogenology. 2021;172:193-9. PMid:34246165.

Suzuki K, Eriksson B, Shimizu H, Nagai T, Rodriguez-Martinez H. Effect of hyaluronan on monospermic penetration of porcine oocytes fertilized in vitro. Int J Androl. 2000;23(1):13-21. PMid:10632757.

Teixeira PP, Padilha LC, Oliveira ME, Motheo TF, Silva AS, Barros FF, Coutinho LN, Flôres FN, Lopes MC, Bandarra MB, Silva MA, Vasconcelos RO, Rodrigues LF, Vicente WR. Laparoscopic ovum collection in sheep: gross and microscopic evaluation of the ovary and influence on oocyte production. Anim Reprod Sci. 2011;127(3-4):169-75. PMid:21907507.

Telford NA, Watson AJ, Schultz GA. Transition from maternal to embryonic control in early mammalian development: a comparison of several species. Mol Reprod Dev. 1990;26(1):90-100. PMid:2189447.

Tervit HR, Whittingham DG, Rowson LE. Successful culture in vitro of sheep and cattle ova. J Reprod Fertil. 1972;30(3):493-7. PMid:4672493.

Thompson JG, Brown HM, Sutton-McDowall ML. Measuring embryo metabolism to predict embryo quality. Reprod Fertil Dev. 2016;28(2):41-50. PMid:27062873.

Thompson JG, Gardner DK, Pugh PA, McMillan WH, Tervit HR. Lamb birth weight is affected by culture system utilized during in vitro pre-elongation development of ovine embryos. Biol Reprod. 1995;53(6):1385-91. PMid:8562695.

Thompson JG, Sherman AN, Allen NW, McGowan LT, Tervit HR. Total protein content and protein synthesis within pre-elongation stage bovine embryos. Mol Reprod Dev. 1998;50(2):139-45.<139::AID-MRD3>3.0.CO;2-L. PMid:9590529.

Thompson JG. Defining the requirements for bovine embryo culture. Theriogenology. 1996;45(1):27-40.

Thompson JG. In vitro culture and embryo metabolism of cattle and sheep embryos - a decade of achievement. Anim Reprod Sci. 2000;60-61:263-75. PMid:10844200.

Tian H, Liu K, Zhang Y, Qi Q, Wang C, Guan H, Yan F, Hou J. Adult follicular fluid supplementation during in vitro maturation improves the developmental competence of prepubertal lamb oocytes. Theriogenology. 2019;130:157-62. PMid:30913500.

Tibary A, Anouassi A, Khatir H. Update on reproductive biotechnologies in small ruminants and camelids. Theriogenology. 2005;64(3):618-38. PMid:15964064.

Toyoda Y, Sato E, Naito K. Role of the cumulus oophorus in mammalian fertilization. In: Mohri H, Takahashi M, Tachi C, editors. Biology of the germ line. Basel: Karger Publishers; 1993. p. 111-24.

Van Soom A, Tanghe S, De Pauw I, Maes D, de Kruif A. Function of the cumulus oophorus before and during mammalian fertilization. Reprod Domest Anim. 2002;37(3):144-51. PMid:12071888.

Vandevoort CA, Cherr GN, Overstreet JW. Hyaluronic acid enhances the zona pellucida-induced acrosome reaction of macaque sperm. J Androl. 1997;18(1):1-5. PMid:9089061.

Vázquez MI, Forcada F, Casao A, Abecia JA, Sosa C, Palacín I. Undernutrition and exogenous melatonin can affect the in vitro developmental competence of ovine oocytes on a seasonal basis. Reprod Domest Anim. 2010;45(4):677-84. PMid:19281597.

Veshkini A, Asadi H, Khadem AA, Mohammadi-Sangcheshmeh A, Khazabi S, Aminafshar M, Deldar H, Soleimani M, Cinar MU. Effect of Linolenic acid during in vitro maturation of ovine oocytes: embryonic developmental potential and mRNA abundances of genes involved in apoptosis. J Assist Reprod Genet. 2015;32(4):653-9. PMid:25715790.

Viana JHM [homepage on the Internet]. Champaign: International Embryo Technology Society; 2022 [cited 2023 Jan 30]. Available from:

Wan PC, Hao ZD, Zhou P, Wu Y, Yang L, Cui MS, Liu SR, Zeng SM. Effects of SOF and CR1 media on developmental competence and cell apoptosis of ovine in vitro fertilization embryos. Anim Reprod Sci. 2009;114(1-3):279-88. PMid:19008058.

Wang DC, Huang JC, Lo NW, Chen LR, Mermillod P, Ma WL, Chiang HI, Ju JC. Sonic Hedgehog promotes in vitro oocyte maturation and term development of embryos in Taiwan native goats. Theriogenology. 2017;103:52-8. PMid:28779609.

Wang L, Jiang X, Wu Y, Lin J, Zhang L, Yang N, Huang J. Effect of milrinone on the developmental competence of growing lamb oocytes identified with brilliant cresyl blue. Theriogenology. 2016;86(8):2020-7. PMid:27481813.

Wang Z, Lin P, Yu S. Effects of ghrelin on developmental competence and gene expression of in vitro fertilized ovine embryos. Theriogenology. 2013;79(4):695-701. PMid:23290751.

Widayati DT, Pangestu M. Effect of follicle-stimulating hormone on Bligon goat oocyte maturation and embryonic development post in vitro fertilization. Vet World. 2020;13(11):2443-6. PMid:33363339.

Yanagimachi R. Mysteries and unsolved problems of mammalian fertilization and related topics. Biol Reprod. 2022;106(4):644-75. PMid:35292804.

Zhang L, Jiang S, Wozniak PJ, Yang X, Godke RA. Cumulus cell function during bovine oocyte maturation, fertilization, and embryo development in vitro. Mol Reprod Dev. 1995;40(3):338-44. PMid:7772344.

Zhou P, Wu YG, Li Q, Lan GC, Wang G, Gao D, Tan JH. The interactions between cysteamine, cystine and cumulus cells increase the intracellular glutathione level and developmental capacity of goat cumulus-denuded oocytes. Reproduction. 2008;135(5):605-11. PMid:18411409.

Zhu J, Moawad AR, Wang CY, Li HF, Ren JY, Dai YF. Advances in in vitro production of sheep embryos. Int J Vet Sci Med. 2018;6(Suppl 1):S15-26. PMid:30761316.

Submitted date:

Accepted date:

654bd66fa953954b9e71b0b2 animreprod Articles
Links & Downloads

Anim Reprod

Share this page
Page Sections