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Home / Equine Research Bank / Reprod Biol Endocrinol / Effect of holding equine oocytes in meiosis inhibitor-free m...
Reproductive biology and endocrinology : RB&E2014; 12; 99; doi: 10.1186/1477-7827-12-99

Effect of holding equine oocytes in meiosis inhibitor-free medium before in vitro maturation and of holding temperature on meiotic suppression and mitochondrial energy/redox potential.

Abstract: Evaluation of mitochondrial function offers an alternative to evaluate embryo development for assessment of oocyte viability, but little information is available on the relationship between mitochondrial and chromatin status in equine oocytes. We evaluated these parameters in immature equine oocytes either fixed immediately (IMM) or held overnight in an Earle's/Hank's' M199-based medium in the absence of meiotic inhibitors (EH treatment), and in mature oocytes. We hypothesized that EH holding may affect mitochondrial function and that holding temperature may affect the efficiency of meiotic suppression. Methods: Experiment 1 - Equine oocytes processed immediately or held in EH at uncontrolled temperature (22 to 27°C) were evaluated for initial chromatin configuration, in vitro maturation (IVM) rates and mitochondrial energy/redox potential. Experiment 2 - We then investigated the effect of holding temperature (25°C, 30°C, 38°C) on initial chromatin status of held oocytes, and subsequently repeated mitochondrial energy/redox assessment of oocytes held at 25°C vs. immediately-evaluated controls. Results: EH holding at uncontrolled temperature was associated with advancement of germinal vesicle (GV) chromatin condensation and with meiotic resumption, as well as a lower maturation rate after IVM. Holding did not have a significant effect on mitochondrial distribution within chromatin configurations. Independent of treatment, oocytes having condensed chromatin had a significantly higher proportion of perinuclear/pericortical mitochondrial distribution than did other GV configurations. Holding did not detrimentally affect oocyte energy/redox parameters in viable GV-stage oocytes. There were no significant differences in chromatin configuration between oocytes held at 25°C and controls, whereas holding at higher temperature was associated with meiosis resumption and loss of oocytes having the condensed chromatin GV configuration. Holding at 25°C was not associated with progression of mitochondrial distribution pattern and there were no significant differences in oocyte energy/redox parameters between these oocytes and controls. Conclusions: Mitochondrial distribution in equine GV-stage oocytes is correlated with chromatin configuration within the GV. Progression of chromatin configuration and mitochondrial status during holding are dependent on temperature. EH holding at 25°C maintains meiotic arrest, viability and mitochondrial potential of equine oocytes. This is the first report on the effects of EH treatment on oocyte mitochondrial energy/redox potential.
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Publication Date: 2014-10-11 PubMed ID: 25306508PubMed Central: PMC4209075DOI: 10.1186/1477-7827-12-99Google Scholar: Lookup
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  • Journal Article
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  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research deals with the effect of holding equine oocytes (horse egg cells) in a specific medium before in vitro maturation, and the impact of different holding temperatures on meiotic suppression and mitochondrial function. The study suggests that these factors may affect whether or not these oocytes are viable.

Research Process

  • The scientists examined how holding immature equine oocytes in an Earle’s/Hank’s M199-based medium, devoid of meiotic inhibitors, impacts both chromatin and mitochondrial status, specifically the mitochondrial energy/redox potential.
  • The oocytes were either fixed immediately or held overnight with the experiment divided into two parts.
  • In the first experiment, the oocytes were held at an uncontrolled temperature (22 to 27°C) while in the second experiment, they were held at controlled temperatures of 25°C, 30°C, and 38°C.
  • After each experiment, the oocytes were evaluated for initial chromatin configuration, in vitro maturation (IVM) rates, and mitochondrial energy/redox potential.

Findings

  • The results showed that holding the oocytes at an uncontrolled temperature led to advancement of germinal vesicle (GV) chromatin condensation and meiotic resumption, and a reduced maturation rate post in vitro maturation (IVM).
  • The holding process did not significantly affect the distribution of mitochondria within various chromatin configurations.
  • Oocytes demonstrating the condensed form of chromatin had a higher proportion of mitochondria located near the nucleus (perinuclear) or towards the outer layer (pericortical) compared to other GV configurations.
  • The holding did not negatively impact the energy/redox parameters of viable GV-stage oocytes.

Temperature Specific Findings

  • When analyzing the effects of fixed holding temperatures, there were no significant differences in chromatin configuration between oocytes held at 25°C and the control group.
  • Holding the oocytes at higher temperatures resulted in the initiation of meiosis and a loss of oocytes presenting the condensed chromatin GV configuration.
  • Keeping the oocytes at 25°C did not lead to progression in the mitochondrial distribution pattern, and there were no significant differences in the oocyte energy/redox parameters between the 25°C samples and the control group.

Conclusions

  • The results of this research imply that in equine GV-stage oocytes, their chromatin configuration within the GV is correlated to the distribution of mitochondria.
  • Also, the progression of chromatin configuration and mitochondrial status while holding these oocytes is temperature dependent.
  • Lastly, it was found that holding at 25°C maintains meiotic arrest, as well as the viability and mitochondrial potential of equine oocytes, indicating that 25°C could be a suitable temperature for holding oocytes in the absence of meiotic inhibitors.

Cite This Article

APA
Martino NA, Dell'Aquila ME, Filioli Uranio M, Rutigliano L, Nicassio M, Lacalandra GM, Hinrichs K. (2014). Effect of holding equine oocytes in meiosis inhibitor-free medium before in vitro maturation and of holding temperature on meiotic suppression and mitochondrial energy/redox potential. Reprod Biol Endocrinol, 12, 99. https://doi.org/10.1186/1477-7827-12-99

Publication

Reproductive biology and endocrinology : RB&E
ISSN: 1477-7827
NlmUniqueID: 101153627
Country: England
Language: English
Volume: 12
Pages: 99
PII: 99

Researcher Affiliations

Martino, Nicola A
  • Veterinary Clinics and Animal Productions Unit-Dipartimento dell'Emergenza e Trapianti D'Organo (DETO), Università di Bari Aldo Moro, Str, Prov, Casamassima Km 3°, Valenzano 70010, Bari, Italy. nicmartino@libero.it.
Dell'Aquila, Maria E
    Filioli Uranio, Manuel
      Rutigliano, Lucia
        Nicassio, Michele
          Lacalandra, Giovanni M
            Hinrichs, Katrin

              MeSH Terms

              • Abattoirs
              • Animals
              • Cell Survival
              • Chromatin Assembly and Disassembly
              • Cold Temperature / adverse effects
              • Culture Media
              • Energy Metabolism
              • Female
              • Horses / physiology
              • In Vitro Oocyte Maturation Techniques / veterinary
              • Meiosis
              • Microscopy, Confocal / veterinary
              • Microscopy, Fluorescence / veterinary
              • Mitochondria / metabolism
              • Oocytes / cytology
              • Oocytes / metabolism
              • Oogenesis
              • Oxidation-Reduction
              • Reactive Oxygen Species / metabolism

              References

              This article includes 48 references
              1. Choi YH, Love LB, Varner DD, Hinrichs K. Holding immature equine oocytes in the absence of meiotic inhibitors: effect on germinal vesicle chromatin and blastocyst development after intracytoplasmic sperm injection.. Theriogenology 2006 Sep 1;66(4):955-63.
                doi: 10.1016/j.theriogenology.2006.01.064pubmed: 16574209google scholar: lookup
              2. Choi YH, Love CC, Varner DD, Hinrichs K. Equine blastocyst development after intracytoplasmic injection of sperm subjected to two freeze-thaw cycles.. Theriogenology 2006 Mar 1;65(4):808-19.
                doi: 10.1016/j.theriogenology.2005.04.035pubmed: 16095679google scholar: lookup
              3. Choi YH, Love LB, Varner DD, Hinrichs K. Effect of holding technique and culture drop size in individual or group culture on blastocyst development after ICSI of equine oocytes with low meiotic competence.. Anim Reprod Sci 2007 Nov;102(1-2):38-47.
                doi: 10.1016/j.anireprosci.2006.09.028pubmed: 17088030google scholar: lookup
              4. Ribeiro BI, Love LB, Choi YH, Hinrichs K. Transport of equine ovaries for assisted reproduction.. Anim Reprod Sci 2008 Oct;108(1-2):171-9.
                doi: 10.1016/j.anireprosci.2007.08.001pubmed: 17888596google scholar: lookup
              5. Choi YH, Hartman DL, Fissore RA, Bedford-Guaus SJ, Hinrichs K. Effect of sperm extract injection volume, injection of PLCzeta cRNA, and tissue cell line on efficiency of equine nuclear transfer.. Cloning Stem Cells 2009 Jun;11(2):301-8.
                doi: 10.1089/clo.2008.0077pubmed: 19508114google scholar: lookup
              6. Hinrichs K, Choi YH, Norris JD, Love LB, Bedford-Guaus SJ, Hartman DL, Velez IC. Evaluation of foal production following intracytoplasmic sperm injection and blastocyst culture of oocytes from ovaries collected immediately before euthanasia or after death of mares under field conditions.. J Am Vet Med Assoc 2012 Oct 15;241(8):1070-4.
                doi: 10.2460/javma.241.8.1070pubmed: 23039983google scholar: lookup
              7. Choi YH, Norris JD, Velez IC, Jacobson CC, Hartman DL, Hinrichs K. A viable foal obtained by equine somatic cell nuclear transfer using oocytes recovered from immature follicles of live mares.. Theriogenology 2013 Mar 15;79(5):791-6.e1.
                doi: 10.1016/j.theriogenology.2012.12.005pubmed: 23312717google scholar: lookup
              8. Foss R, Ortis H, Hinrichs K. Effect of potential oocyte transport protocols on blastocyst rates after intracytoplasmic sperm injection in the horse.. Equine Vet J Suppl 2013 Dec;(45):39-43.
                doi: 10.1111/evj.12159pubmed: 24304402google scholar: lookup
              9. Hinrichs K, Choi YH, Love LB, Varner DD, Love CC, Walckenaer BE. Chromatin configuration within the germinal vesicle of horse oocytes: changes post mortem and relationship to meiotic and developmental competence.. Biol Reprod 2005 May;72(5):1142-50.
                doi: 10.1095/biolreprod.104.036012pubmed: 15647456google scholar: lookup
              10. Sirard MA, Richard F, Mayes M. Controlling meiotic resumption in bovine oocytes: a review.. Theriogenology 1998 Jan 15;49(2):483-97.
                doi: 10.1016/S0093-691X(97)00420-2pubmed: 10732029google scholar: lookup
              11. Lonergan P, Khatir H, Carolan C, Mermillod P. Bovine blastocyst production in vitro after inhibition of oocyte meiotic resumption for 24 h.. J Reprod Fertil 1997 Mar;109(2):355-65.
                doi: 10.1530/jrf.0.1090355pubmed: 9155746google scholar: lookup
              12. Love CC, Love LB, Varner DD, Hinrichs K. Effect of holding at room temperature on initial chromatin configuration and in vitro maturation rate of equine oocytes.. Theriogenology 2002 May;57(8):1973-9.
                doi: 10.1016/S0093-691X(02)00646-5pubmed: 12066858google scholar: lookup
              13. Zeng HT, Ren Z, Guzman L, Wang X, Sutton-McDowall ML, Ritter LJ, De Vos M, Smitz J, Thompson JG, Gilchrist RB. Heparin and cAMP modulators interact during pre-in vitro maturation to affect mouse and human oocyte meiosis and developmental competence.. Hum Reprod 2013 Jun;28(6):1536-45.
                doi: 10.1093/humrep/det086pubmed: 23559189google scholar: lookup
              14. Dell'Aquila ME, Albrizio M, Maritato F, Minoia P, Hinrichs K. Meiotic competence of equine oocytes and pronucleus formation after intracytoplasmic sperm injection (ICSI) as related to granulosa cell apoptosis.. Biol Reprod 2003 Jun;68(6):2065-72.
                doi: 10.1095/biolreprod.102.009852pubmed: 12606481google scholar: lookup
              15. Mortensen CJ, Choi YH, Ing NH, Kraemer DC, Vogelsang MM, Hinrichs K. Heat shock protein 70 gene expression in equine blastocysts after exposure of oocytes to high temperatures in vitro or in vivo after exercise of donor mares.. Theriogenology 2010 Aug;74(3):374-83.
                doi: 10.1016/j.theriogenology.2010.02.020pubmed: 20416934google scholar: lookup
              16. Colleoni S, Barbacini S, Necci D, Duchi R, Lazzari G, Galli C. Application of ovum pick-up, intracytoplasmic sperm injection and embryo culture in equine practice. Proceedings of the American Association of Equine Practitioners 1–5 December 2007; Orlando, Florida. American Association of Equine Practitioners, Lexington. 2007.
              17. Choi YH, Roasa LM, Love CC, Varner DD, Brinsko SP, Hinrichs K. Blastocyst formation rates in vivo and in vitro of in vitro-matured equine oocytes fertilized by intracytoplasmic sperm injection.. Biol Reprod 2004 May;70(5):1231-8.
                doi: 10.1095/biolreprod.103.023903pubmed: 14695908google scholar: lookup
              18. Jacobson CC, Choi YH, Hayden SS, Hinrichs K. Recovery of mare oocytes on a fixed biweekly schedule, and resulting blastocyst formation after intracytoplasmic sperm injection.. Theriogenology 2010 May;73(8):1116-26.
                doi: 10.1016/j.theriogenology.2010.01.013pubmed: 20202674google scholar: lookup
              19. Choi YH, Varner DD, Love CC, Hartman DL, Hinrichs K. Production of live foals via intracytoplasmic injection of lyophilized sperm and sperm extract in the horse.. Reproduction 2011 Oct;142(4):529-38.
                doi: 10.1530/REP-11-0145pubmed: 21846810google scholar: lookup
              20. Alonso A, Baca Castex C, Ferrante A, Pinto M, Castañeira C, Trasorras V, Gambarotta MC, Losinno L, Miragaya M. In vitro equine embryo production using air-dried spermatozoa, with different activation protocols and culture systems.. Andrologia 2015 May;47(4):387-94.
                pubmed: 24684246doi: 10.1111/and.12273google scholar: lookup
              21. Galli C, Duchi R, Colleoni S, Lagutina I, Lazzari G. Ovum pick up, intracytoplasmic sperm injection and somatic cell nuclear transfer in cattle, buffalo and horses: from the research laboratory to clinical practice.. Theriogenology 2014 Jan 1;81(1):138-51.
                doi: 10.1016/j.theriogenology.2013.09.008pubmed: 24274418google scholar: lookup
              22. Zaniboni A, Merlo B, Zannoni A, Bernardini C, Lavitrano M, Forni M, Mari G, Bacci ML. Expression of fluorescent reporter protein in equine embryos produced through intracytoplasmic sperm injection mediated gene transfer (ICSI-MGT).. Anim Reprod Sci 2013 Feb;137(1-2):53-61.
                doi: 10.1016/j.anireprosci.2012.12.010pubmed: 23312469google scholar: lookup
              23. Smits K, Govaere J, Hoogewijs M, Piepers S, Van Soom A. A pilot comparison of laser-assisted vs piezo drill ICSI for the in vitro production of horse embryos.. Reprod Domest Anim 2012 Feb;47(1):e1-3.
                doi: 10.1111/j.1439-0531.2011.01814.xpubmed: 21950451google scholar: lookup
              24. Smits K, Govaere J, Peelman LJ, Goossens K, de Graaf DC, Vercauteren D, Vandaele L, Hoogewijs M, Wydooghe E, Stout T, Van Soom A. Influence of the uterine environment on the development of in vitro-produced equine embryos.. Reproduction 2012 Feb;143(2):173-81.
                doi: 10.1530/REP-11-0217pubmed: 22089531google scholar: lookup
              25. Pereyra-Bonnet F, Fernández-Martín R, Olivera R, Jarazo J, Vichera G, Gibbons A, Salamone D. A unique method to produce transgenic embryos in ovine, porcine, feline, bovine and equine species.. Reprod Fertil Dev 2008;20(7):741-9.
                doi: 10.1071/RD07172pubmed: 18842176google scholar: lookup
              26. Dumollard R, Marangos P, Fitzharris G, Swann K, Duchen M, Carroll J. Sperm-triggered [Ca2+] oscillations and Ca2+ homeostasis in the mouse egg have an absolute requirement for mitochondrial ATP production.. Development 2004 Jul;131(13):3057-67.
                doi: 10.1242/dev.01181pubmed: 15163630google scholar: lookup
              27. Pikó L, Chase DG. Role of the mitochondrial genome during early development in mice. Effects of ethidium bromide and chloramphenicol.. J Cell Biol 1973 Aug;58(2):357-78.
                doi: 10.1083/jcb.58.2.357pmc: PMC2109054pubmed: 4738106google scholar: lookup
              28. Larsson NG, Wang J, Wilhelmsson H, Oldfors A, Rustin P, Lewandoski M, Barsh GS, Clayton DA. Mitochondrial transcription factor A is necessary for mtDNA maintenance and embryogenesis in mice.. Nat Genet 1998 Mar;18(3):231-6.
                doi: 10.1038/ng0398-231pubmed: 9500544google scholar: lookup
              29. Hansford RG. Physiological role of mitochondrial Ca2+ transport.. J Bioenerg Biomembr 1994 Oct;26(5):495-508.
                doi: 10.1007/BF00762734pubmed: 7896765google scholar: lookup
              30. Turrens JF. Mitochondrial formation of reactive oxygen species.. J Physiol 2003 Oct 15;552(Pt 2):335-44.
                doi: 10.1113/jphysiol.2003.049478pmc: PMC2343396pubmed: 14561818google scholar: lookup
              31. Funato Y, Michiue T, Asashima M, Miki H. The thioredoxin-related redox-regulating protein nucleoredoxin inhibits Wnt-beta-catenin signalling through dishevelled.. Nat Cell Biol 2006 May;8(5):501-8.
                doi: 10.1038/ncb1405pubmed: 16604061google scholar: lookup
              32. Liu H, Colavitti R, Rovira II, Finkel T. Redox-dependent transcriptional regulation.. Circ Res 2005 Nov 11;97(10):967-74.
                doi: 10.1161/01.RES.0000188210.72062.10pubmed: 16284189google scholar: lookup
              33. Rahman I, Marwick J, Kirkham P. Redox modulation of chromatin remodeling: impact on histone acetylation and deacetylation, NF-kappaB and pro-inflammatory gene expression.. Biochem Pharmacol 2004 Sep 15;68(6):1255-67.
                doi: 10.1016/j.bcp.2004.05.042pubmed: 15313424google scholar: lookup
              34. Banrezes B, Sainte-Beuve T, Canon E, Schultz RM, Cancela J, Ozil JP. Adult body weight is programmed by a redox-regulated and energy-dependent process during the pronuclear stage in mouse.. PLoS One 2011;6(12):e29388.
                doi: 10.1371/journal.pone.0029388pmc: PMC3247262pubmed: 22216268google scholar: lookup
              35. Raval J, Lyman S, Nitta T, Mohuczy D, Lemasters JJ, Kim JS, Behrns KE. Basal reactive oxygen species determine the susceptibility to apoptosis in cirrhotic hepatocytes.. Free Radic Biol Med 2006 Dec 1;41(11):1645-54.
                doi: 10.1016/j.freeradbiomed.2006.07.023pmc: PMC1773006pubmed: 17145552google scholar: lookup
              36. Wakefield SL, Lane M, Schulz SJ, Hebart ML, Thompson JG, Mitchell M. Maternal supply of omega-3 polyunsaturated fatty acids alter mechanisms involved in oocyte and early embryo development in the mouse.. Am J Physiol Endocrinol Metab 2008 Feb;294(2):E425-34.
                doi: 10.1152/ajpendo.00409.2007pubmed: 18073322google scholar: lookup
              37. Martino NA, Lacalandra GM, Filioli Uranio M, Ambruosi B, Caira M, Silvestre F, Pizzi F, Desantis S, Accogli G, Dell'Aquila ME. Oocyte mitochondrial bioenergy potential and oxidative stress: within-/between-subject, in vivo versus in vitro maturation, and age-related variations in a sheep model.. Fertil Steril 2012 Mar;97(3):720-8.e1.
                doi: 10.1016/j.fertnstert.2011.12.014pubmed: 22260855google scholar: lookup
              38. Dumollard R, Duchen M, Sardet C. Calcium signals and mitochondria at fertilisation.. Semin Cell Dev Biol 2006 Apr;17(2):314-23.
                doi: 10.1016/j.semcdb.2006.02.009pubmed: 16574440google scholar: lookup
              39. Torner H, Alm H, Kanitz W, Goellnitz K, Becker F, Poehland R, Bruessow KP, Tuchscherer A. Effect of initial cumulus morphology on meiotic dynamic and status of mitochondria in horse oocytes during IVM.. Reprod Domest Anim 2007 Apr;42(2):176-83.
                doi: 10.1111/j.1439-0531.2006.00749.xpubmed: 17348975google scholar: lookup
              40. Ambruosi B, Lacalandra GM, Iorga AI, De Santis T, Mugnier S, Matarrese R, Goudet G, Dell'aquila ME. Cytoplasmic lipid droplets and mitochondrial distribution in equine oocytes: Implications on oocyte maturation, fertilization and developmental competence after ICSI.. Theriogenology 2009 Apr 15;71(7):1093-104.
                doi: 10.1016/j.theriogenology.2008.12.002pubmed: 19167745google scholar: lookup
              41. Ambruosi B, Uranio MF, Sardanelli AM, Pocar P, Martino NA, Paternoster MS, Amati F, Dell'Aquila ME. In vitro acute exposure to DEHP affects oocyte meiotic maturation, energy and oxidative stress parameters in a large animal model.. PLoS One 2011;6(11):e27452.
                doi: 10.1371/journal.pone.0027452pmc: PMC3208636pubmed: 22076161google scholar: lookup
              42. Dell'Aquila ME, Masterson M, Maritato F, Hinrichs K. Influence of oocyte collection technique on initial chromatin configuration, meiotic competence, and male pronucleus formation after intracytoplasmic sperm injection (ICSI) of equine oocytes.. Mol Reprod Dev 2001 Sep;60(1):79-88.
                doi: 10.1002/mrd.1064pubmed: 11550271google scholar: lookup
              43. Zinchuk V, Grossenbacher-Zinchuk O. Recent advances in quantitative colocalization analysis: focus on neuroscience.. Prog Histochem Cytochem 2009;44(3):125-72.
                doi: 10.1016/j.proghi.2009.03.001pubmed: 19822255google scholar: lookup
              44. Vernunft A, Alm H, Tuchscherer A, Kanitz W, Hinrichs K, Torner H. Chromatin and cytoplasmic characteristics of equine oocytes recovered by transvaginal ultrasound-guided follicle aspiration are influenced by the developmental stage of their follicle of origin.. Theriogenology 2013 Jul 1;80(1):1-9.
                doi: 10.1016/j.theriogenology.2013.03.023pubmed: 23623162google scholar: lookup
              45. Hinrichs K, Schmidt AL. Meiotic competence in horse oocytes: interactions among chromatin configuration, follicle size, cumulus morphology, and season.. Biol Reprod 2000 May;62(5):1402-8.
                doi: 10.1095/biolreprod62.5.1402pubmed: 10775193google scholar: lookup
              46. Parfenov V, Potchukalina G, Dudina L, Kostyuchek D, Gruzova M. Human antral follicles: oocyte nucleus and the karyosphere formation (electron microscopic and autoradiographic data).. Gamete Res 1989 Feb;22(2):219-31.
                doi: 10.1002/mrd.1120220209pubmed: 2468591google scholar: lookup
              47. Hinrichs K, Williams KA. Relationships among oocyte-cumulus morphology, follicular atresia, initial chromatin configuration, and oocyte meiotic competence in the horse.. Biol Reprod 1997 Aug;57(2):377-84.
                doi: 10.1095/biolreprod57.2.377pubmed: 9241053google scholar: lookup
              48. Alm H, Choi YH, Love L, Heleil B, Torner H, Hinrichs K. Holding bovine oocytes in the absence of maturation inhibitors: kinetics of in vitro maturation and effect on blastocyst development after in vitro fertilization.. Theriogenology 2008 Oct 15;70(7):1024-9.
                doi: 10.1016/j.theriogenology.2008.05.062pubmed: 18644621google scholar: lookup

              Citations

              This article has been cited 2 times.
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              2. Salgado RM, Brom-de-Luna JG, Resende HL, Canesin HS, Hinrichs K. Lower blastocyst quality after conventional vs. Piezo ICSI in the horse reflects delayed sperm component remodeling and oocyte activation.. J Assist Reprod Genet 2018 May;35(5):825-840.
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