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Prospective randomized multicentre comparison on sibling oocytes comparing G-Series media system with antioxidants versus standard G-Series media system

Published:February 05, 2020DOI:https://doi.org/10.1016/j.rbmo.2020.01.026

      Abstract

      Research question

      Does the inclusion of three antioxidants (A3), acetyl-l-carnitine (ALC), N-acetyl-l-cysteine (NAC) and alpha-lipoic acid (ALA) improve human embryo development and pregnancy potential?

      Design

      Prospective randomized multicentre comparison of sibling oocytes. A total of 1563 metaphase II oocytes from 133 patients in two IVF centres. Day 3 embryo and day 5/6 blastocyst quality were assessed. Good embryo quality on day 3 was defined as 8 to 10 cells with even cells and low fragmentation; good quality blastocysts as 3BB or greater. Clinical outcome was assessed on transfers of fresh or vitrified–warmed blastocyst on day 5.

      Results

      Of the two-pronuclei, 40.7% (G-Series) and 50.2% (G-Series with A3 group) resulted in good quality embryos on day 3 (P < 0.05). The implantation rate by fetal sac was 39.2% and 50.6%, and by fetal heartbeat was 37.8% and 47.1% for the G-Series and G-Series with A3 group, respectively. When stratified by female patient age, patients 35–40 years had an implantation rate by fetal sac and heart of 23.5% in the G-Series compared with 57.5% (P < 0.05) and 50.0% (P < 0.05) in the A3 group. The ongoing pregnancies in patients 35–40 years were significantly higher in the A3 group (50%) compared with the control (25.8%) (P < 0.05).

      Conclusions

      The presence of antioxidants during IVF and embryo culture for patients 35–40 years resulted in a significant increase in implantation and pregnancy rate. Supplementation of antioxidants to IVF and culture media may therefore improve the viability of human embryos in assisted reproductive technologies, plausibly through the reduction of oxidative stress.

      Keywords

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      References

        • Agarwal A.
        • Aponte-Mellado A.
        • Premkumar B.J.
        • Shaman A.
        • Gupta S.
        The effects of oxidative stress on female reproduction: a review.
        Reprod. Biol. Endocrinol. 2012; 10: 49
        • Agarwal A.
        • Majzoub A.
        Role of Antioxidants in Assisted Reproductive Techniques.
        World J. Mens Health. 2017; 35: 77-93
        • Aitken R.J.
        Reactive oxygen species as mediators of sperm capacitation and pathological damage.
        Mol. Reprod. Dev. 2017; 84: 1039-1052
        • Aitken R.J.
        • Baker M.A.
        • Sawyer D.
        Oxidative stress in the male germ line and its role in the aetiology of male infertility and genetic disease.
        Reprod. BioMed. Online. 2003; 7: 65-70
        • Aliciguzel Y.
        • Aslan M.
        N-acetyl cysteine, L-cysteine, and beta-mercaptoethanol augment selenium-glutathione peroxidase activity in glucose-6-phosphate dehydrogenase-deficient human erythrocytes.
        Clin. Exp. Med. 2004; 4: 50-55
        • Alpha Scientists in Reproductive Medicine and ESHRE Special Interest Group Embryology
        Istanbul consensus workshop on embryo assessment: proceedings of an expert meeting.
        Reprod. BioMed. Online. 2011; 22: 632-646
        • Andrae U.
        • Singh J.
        • Ziegler-Skylakakis K.
        Pyruvate and related alpha-ketoacids protect mammalian cells in culture against hydrogen peroxide-induced cytotoxicity.
        Toxicol. Lett. 1985; 28: 93-98
        • Awonuga A.O.
        • Yang Y.
        • Rappolee D.A.
        When stresses collide.
        Biology of Reproduction. 2013; 89: 1-2
        • Banihani S.
        • Sharma R.
        • Bayachou M.
        • Sabanegh E.
        • Agarwal A
        Human sperm DNA oxidation, motility and viability in the presence of L-carnitine duringin vitro incubation and centrifugation.
        Andrologia. 2012; 44: 505-512
        • Bavister BD.
        Culture of preimplantation embryos: facts and artefacts.
        Hum. Reprod. Update. 1995; 1: 91-148
        • Belli M.
        • Zhang L.
        • Liu X.
        • Donjacour A.
        • Ruggeri E.
        • Palmerini M.G.
        • Nottola S.A.
        • Macchiarelli G.
        • Rinaudo P.
        Oxygen concentration alters mitochondrial structure and function inin vitro fertilized preimplantation mouse embryos.
        Hum. Reprod. 2019; 34: 601-611
        • Ben-Meir A.
        • Burstein E.
        • Borrego-Alvarez A.
        • Chong J.
        • Wong E.
        • Yavorska T.
        • Naranian T.
        • Chi M.
        • Wang Y.
        • Bentov Y.
        • Alexis J.
        • Meriano J.
        • Sung H-K.
        • Gasser D.L.
        • Moley K.H.
        • Hekimi S.
        • Casper R.F.
        • Juriscova A
        Coenzyme Q10 restores oocyte mitochondrial function and fertility during reproductive ageing.
        Ageing Cell. 2015; 14: 887-895
        • Bilska A.
        • Wlodek L.
        Lipoic acid–the drug of the future?.
        Pharmacol. Rep. 2005; 57: 570-577
        • Bontekoe S.
        • Mantikou E.
        • van Wely M.
        • Seshadri S.
        • Repping S.
        • Mastenbroek S.
        Low oxygen concentrations for embryo culture in assisted reproductive technologies.
        The Cochrane database of systematic reviews. 2012; 7CD008950
        • Bui A.D.
        • Sharma R.
        • Henkel R.
        • Agarwal A.
        Reactive oxygen species impact on sperm DNA and its role in male infertility.
        Andrologia. 2018; 50: e13012
        • Caamano J.N.
        • Ryoo Z.Y.
        • Youngs C.R.
        Promotion of development of bovine embryos producedin vitro by addition of cysteine and beta-mercaptoethanol to a chemically defined culture system.
        J. Dairy Sci. 1998; 81: 369-374
        • de Matos D.G.
        • Furnus C.C.
        • Moses D.F.
        • Martinez A.G.
        • Matkovic M.
        Stimulation of glutathione synthesis ofin vitro matured bovine oocytes and its effect on embryo development and freezability.
        Mol. Reprod. Dev. 1996; 45: 451-457
        • de Matos D.G.
        • Nogueira D.
        • Cortvrindt R.
        • Herrera C.
        • Adriaenssens T.
        • Pasqualini R.S.
        • Smitz J.
        Capacity of adult and prepubertal mouse oocytes to undergo embryo development in the presence of cysteamine.
        Mol. Reprod. Dev. 2003; 64: 214-218
        • Eichenlaub-Ritter U
        Oocyte ageing and its cellualr basis.
        Int. J. Dev. Biol. 2012; 56: 841-852
        • Franasiak J.M.
        • Formann E.J.
        • Hong K.H.
        • Werner M.D.
        • Upham K.M.
        • Treff N.R.
        • Scott R.T.
        Aneuploidy across individual chromosomes at the embryonic level in trophectoderm biopsies: changes with patient age and chromosome structure.
        J Assist Reprod. Genet. 2014; 31: 1501-1509
        • Gardiner C.S.
        • Salmen J.J.
        • Brandt C.J.
        • Stover S.K.
        Glutathione is present in reproductive tract secretions and improves development of mouse embryos after chemically induced glutathione depletion.
        Biol. Reprod. 1998; 59: 431-436
        • Gardner D.K.
        • Lane M.
        Towards a single embryo transfer.
        Reprod. Biomed. Online. 2003; 6: 470-481
        • Gardner D.K.
        • Kelley R.L.
        Impact of the IVF laboratory environment on human preimplantation embryo phenotype.
        J. Dev. Orig. Health Dis. 2017; 8: 418-435
        • Gardner D.K.
        • Lane M.
        Ex vivo early embryo development and effects on gene expression and imprinting.
        Reprod. Fertil. Dev. 2005; 17: 361-370
        • Gardner D.K.
        • Lane M
        Embryo culture systems.
        in: Gardner D.K. Simon C. Handbook ofin vitro fertilization. 4th Edition. CRC press, Boca raton2017: 205-244
        • Gardner D.K.
        • Lane M.
        Culture systems for the human embryo.
        in: Gardner D.K. Weissman A. Howles C. Shoham Z. Textbook of Assisted Reproductive Techniques. 5th Edition. CRC Press, Boca Raton, Florida2018: 200-224
        • Gardner D.K.
        • Lane M.
        • Stevens J.
        • Schlenker T.
        • Schoolcraft W.B
        Blastocyst score affects implantation and pregnancy outcome: towards a single blastocyst transfer.
        Fertil. Steril. 2000; 73: 1155-1158
        • Gardner D.K.
        • Schoolcraft W.B.
        • Wagley L.
        • Schlenker T.
        • Stevens J.
        • Hesla J.
        A prospective randomized trial of blastocyst culture and transfer in in-vitro fertilization.
        Hum. Reprod. 1998; 13: 3434-3440
        • Gaspar R.C.
        • Arnold D.R.
        • Correa C.A.
        • da Rocha Jr., C.V.
        • Penteado J.C.
        • Del Collado M.
        • Vantini R.
        • Garcia J.M.
        • Lopes F.L.
        Oxygen tension affects histone remodeling ofin vitro-produced embryos in a bovine model.
        Theriogenology. 2015; 83: 1408-1415
        • Ghosh J.
        • Coutifaris C.
        • Sapienza C.
        • Mainigi M.
        Global DNA methylation levels are altered by modifiable clinical manipulations in assisted reproductive technologies.
        Clin. Epigenetics. 2017; 9: 14
        • Hamanaka R.B.
        • Chandel N.C
        Mitochondrial reactive oxygen species regulate cellualr signaling and dictate biological outcomes.
        Trends Biochem. Sci. 2010; 35: 505-513
        • Harvey A.J.
        • Kind K.L.
        • Pantaleon M.
        • Armstrong D.T.
        • Thompson J.G.
        Oxygen-regulated gene expression in bovine blastocysts.
        Biol. Reprod. 2004; 71: 1108-1119
        • Igarashi H.
        • Takahashi T.
        • Nagase S.
        Oocyte ageing underlies female reproductive ageing: biological mechanisms and therapeutic strategies.
        Reprod. Med. Biol. 2015; 14: 159-169
        • Katz-Jaffe M.G
        • Linck D.W.
        • Schoolcraft W.B.
        • Gardner D.K.
        A proteomic analysis of mammalian preimplantation embryonic development.
        Reproduction. 2005; 130: 899-905
        • Kelley R.L.
        • Gardner D.K.
        Combined effects of individual culture and atmospheric oxygen on preimplantation mouse embryosin vitro.
        Reprod. BioMed. Online. 2016; 33: 537-549
        • Kelley R.L.
        • Gardner DK.
        In vitro culture of individual mouse preimplantation embryos: the role of embryo density, microwells, oxygen, timing and conditioned media.
        Reprod. BioMed. Online. 2017; 34: 441-454
        • Kelley R.L.
        • Gardner D.K
        Individual culture and atmospheric oxygen during culture affect mouse preimplantation embryo metabolism and post-implantation development.
        Reprod. BioMed. Online. 2019; 39: 3-18
        • Khurana N.K.
        • Wales R.G.
        Effects of oxygen concentration on the metabolism of [U-14C]glucose by mouse morulae and early blastocystsin vitro.
        Reprod. Fertil. Dev. 1989; 1: 99-106
        • Kind K.L.
        • Collett R.A.
        • Harvey A.J.
        • Thompson J.G.
        Oxygen-regulated expression of GLUT-1, GLUT-3, and VEGF in the mouse blastocyst.
        Mol. Reprod. Dev. 2005; 70: 37-44
        • Kirkegaard K.
        • Hindkjaer J.J.
        • Ingerslev H.J.
        Effect of oxygen concentration on human embryo development evaluated by time-lapse monitoring.
        Fertil. Steril. 2013; 99 (e734): 738-744
        • Kouridakis K.
        • Gardner D.K.
        Pyruvate in embryo culture embryo media acts as an antioxidant.
        in: Proc. Fert. Soc. Aus., Melbourne, Victoria1995: 29 (Abstract)
        • Lees J.G.
        • Gardner D.K.
        • Harvey A.J
        Pluripotent stem cell metabolism and mitochondria: Beyond ATP.
        Stem. Cells Int. 2017; 20172874283
        • Legge M.
        • Sellens M.H.
        Free radical scavengers ameliorate the 2-cell block in mouse embryo culture.
        Hum. Reprod. 1991; 6: 867-871
        • Lewin J.
        • Wells D.
        Preimplantation genetic diagnosis for infertility.
        in: Gardner D.K. Weissman A. Howles C. Shoham Z. Textbook of Assisted Reproductive Techniques. 5th Edition. CRC Press, Boca Raton, Florida2018: 350-358
        • Li W.
        • Goossens K.
        • Van Poucke M.
        • Forier K.
        • Braeckmans K.
        • Van Soom A.
        • Peelman L.J.
        High oxygen tension increases global methylation in bovine 4-cell embryos and blastocysts but does not affect general retrotransposon expression.
        Reprod. Fertil. Dev. 2016; 28: 948-959
        • Lim J.
        • Lious S.S.
        • Hansel W.
        Intracytoplasmic glutathione concentration and beta-mercaptoethanol in preimplantation development of bovine embryo.
        Theriogenology. 1996; 46 (–429): 429
        • Lindenau A.
        • Fischer B.
        Effect of oxygen concentration in the incubator's gas phase on the development of cultured preimplantation rabbit embryos.
        Theriogenology. 1994; 41: 889-898
        • Liu Z.
        • Foote R.H.
        Effects of amino acids on the development of in-vitro matured/in-vitro fertilization bovine embryos in a simple protein-free medium.
        Hum. Reprod. 1995; 10: 2985-2991
        • Meintjes M.
        • Chantilis S.J.
        • Douglas J.D.
        • Rodriguez A.J.
        • Guerami A.R.
        • Bookout D.M.
        • Barnett B.D.
        • Madden J.D.
        A controlled randomized trial evaluating the effect of lowered incubator oxygen tension on live births in a predominantly blastocyst transfer program.
        Hum. Reprod. 2009; 24: 300-307
        • Morbeck D.E.
        • Krisher R.L.
        • Herrick J.R.
        • Baumann N.A.
        • Matern D.
        • Moyer T.
        Composition of commercial media used for human embryo culture.
        Fertil. Steril. 2014; 102 (e759): 759-766
        • Nasr-Esfahani M.H.
        • Johnson M.H.
        Quantitative analysis of cellular glutathione in early preimplantation mouse embryos developingin vivo andin vitro.
        Hum. Reprod. 1992; 7: 1281-1290
        • Ng K.Y.B.
        • Mingels R.
        • Morgan H.
        • Macklon N.
        • Cheong Y.
        In vivo oxygen, temperature and pH dynamics in the female reproductive tract and their importance in human conception: a systematic review.
        Hum. Reprod. Update. 2018; 24: 15-34
        • Noda Y.
        • Matsumoto H.
        • Umaoka Y
        • Tatsumi K.
        • Kishi J.
        • Mori T.
        Involvement of superoxide radicals in the mouse two-cell block.
        Mol. Reprod. Dev. 1991; 28: 356-360
        • O'Fallon J.V.
        • Wright R.W.
        Pyruvate revisited: a non-metabolic role for pyruvate in preimplantation embryo development.
        Theriogenology. 1995; 43: 288
      1. Patel, R., Rinker, L., Peng, J., Chilian, W.M. Reactive oxygen species: The good and the bad. 2018 http://dx.doi.org/10.5772/intechopen.71547

        • Parsanathan R.
        • Jain S.K.
        L-Cysteinein vitro can restore cellular glutathione and inhibits the expression of cell adhesion molecules in G6PD-deficient monocytes.
        Amino. Acids. 2018; 50: 909-921
        • Payne S.R.
        • Munday R.
        • Thompson J.G.
        Addition of superoxide dismutase and catalase does not necessarily overcome developmental retardation of one-cell mouse embryos during in-vitro culture.
        Reprod. Fertil. Dev. 1992; 4: 167-174
        • Perkins A.T.
        • Das T.M.
        • Panzera L.C.
        • Bickel S.E.
        Oxidative stress in oocytes during midprophase induces premature loss of cohesion and chromosome segration errors.
        Proc. Natl. Acad. Sci. USA. 2016; 113: E6823-E6830
        • Perkins A.T.
        • Greig M.M.
        • Sontakke A.A.
        • Peloquin A.S.
        • McPeek M.A.
        • Bickel S.E.
        Increased levels of superoxide dismutase supress meiotic segregation errors in ageing oocytes.
        Chromosma. 2019; 128: 215-222
        • Pool T.B.
        Recent advances in the production of viable human embryosin vitro.
        Reprod. BioMed. Online. 2002; 4: 294-302
        • Rinaudo P.F.
        • Giritharan G.
        • Talbi S.
        • Dobson A.T.
        • Schultz R.M.
        Effects of oxygen tension on gene expression in preimplantation mouse embryos.
        Fertil. Steril. 2006; 86: 1252-1265
        • Sasaki H.
        • Hamatani T.
        • Kamijo S.
        • Iwai M.
        • Kobanawa M.
        • Ogawa S.
        • Miyado K.
        • Tanaka M.
        Impact of oxidative stress on age-associated decline in oocyte developmental competence.
        Front Endocrinol. 2019; 10: 811
        • Showell M.G.
        • Mackenzie-Proctor R.
        • Brown J.
        • Yazdani A.
        • Stankiewicz M.T.
        • Hart RJ.
        Antioxidants for male subfertility.
        Cochrane Database Syst. Rev. 2014; CD007411
        • Showell M.G.
        • Mackenzie-Proctor R.
        • Jordan V.
        • Hart R.J.
        Antioxidants for female subfertility.
        Cochrane Database Syst. Rev. 2017; 7CD007807
        • Swain J.E.
        Optimal human embryo culture.
        Semin. Reprod. Med. 2015; 33: 103-117
        • Swain J.E.
        • Carrell D.
        • Cobo A.
        • Meseguer M.
        • Rubio C.
        • Smith G.D.
        Optimizing the culture environment and embryo manipulation to help maintain embryo developmental potential.
        Fertil. Steril. 2016; 105: 571-587
        • Takahashi M.
        • Nagai T.
        • Hamano S.
        • Kuwayama M.
        • Okamura N.
        • Okano A.
        Effect of thiol compounds onin vitro development and intracellular glutathione content of bovine embryos.
        Biol. Reprod. 1993; 49: 228-232
        • Truong T.
        • Gardner D.K.
        Antioxidants improve IVF outcome and subsequent embryo development in the mouse.
        Hum. Reprod. 2017; 32: 2404-2413
        • Truong T.T.
        • Soh Y.M.
        • Gardner D.K
        Antioxidants improve mouse preimplantation embryo development and viability.
        Hum. Reprod. 2016; 31: 1445-1454
        • Umaoka Y.
        • Noda Y.
        • Narimoto K.
        • Mori T.
        Effects of oxygen toxicity on early development of mouse embryos.
        Mol. Reprod. Dev. 1992; 31: 28-33
        • Wade J.J.
        • MacLachlan V.
        • Kovacs G.
        The success rate of IVF has significantly improved over the last decade.
        Aust. N. Z. J. Obstet. Gynaecol. 2015; 55: 473-476
        • Wale P.L.
        • Gardner D.K.
        Time-lapse analysis of mouse embryo development in oxygen gradients.
        Reprod. BioMed. Online. 2010; 21: 402-410
        • Wale P.L.
        • Gardner D.K.
        Oxygen regulates amino acid turnover and carbohydrate uptake during the preimplantation period of mouse embryo development.
        Biol. Reprod. 2012; 87: 24
        • Wale P.L.
        • Gardner D.K.
        Oxygen affects the ability of mouse blastocysts to regulate ammonium.
        Biol. Reprod. 2013; 89: 75
        • Wale P.L.
        • Gardner D.K.
        The effects of chemical and physical factors on mammalian embryo culture and their importance for the practice of assisted human reproduction.
        Hum. Reprod. Update. 2016; 22: 2-22
        • Yoshida M.
        • Ishigaki K.
        • Nagai T.
        • Chikyu M.
        • Pursel V.G.
        Glutathione concentration during maturation and after fertilization in pig oocytes: relevance to the ability of oocytes to form male pronucleus.
        Biol. Reprod. 1993; 49: 89-94

      Biography

      David Gardner is a Distinguished Professor in the School of BioSciences, University of Melbourne, and the Scientific Director of Melbourne IVF. David has worked in embryology and IVF for over 35 years and maintains an active research programme in embryo physiology and culture, biomarkers, AI, metaboloepigenetics and blastocyst–endometrial signalling.
      Key message
      In a prospective trial, antioxidants in the embryo culture media improved human embryo development in culture and improved transfer outcomes in patients 35 to 40 years old.