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Vol. 63 No. 8: Issue 8

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Morphokinetic analysis of human embryo development and its relationship to the female age: a retrospective time-lapse imaging study

https://doi.org/10.14715/cmb/2017.63.8.18
N. Akhter
Department of Animal Sciences, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan
M. Shahab
Department of Animal Sciences, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan

Corresponding Author(s) : N. Akhter

qancer_75@yahoo.com

Cellular and Molecular Biology, Vol. 63 No. 8: Issue 8

Abstract

Time-lapse imaging technique has provided embryologists with a unique chance of studying the embryo morphokineticsto select the most viable embryos for implantation in the field of IVF (In vitro fertilization).The conventional method of morphological evaluations has proved that maternal age affects the human embryo quality. This retrospective study carried out at Islamabad Clinic Serving Infertile Couples, Islamabad, Pakistan mainly focuses on the effect of female age on human embryo morphokinetics. A total number of 200 patients undergoing ICSI treatment cycles at the clinic were selected for the study and divided into five age groups (< 26, 26-30, 31-35, 36-40, and > 40 years). Embryo culture was done at 37°C, 6% CO2 and 5% oxygen for 5-6 days. Ten time-points were selected for kinetic analysis. The number of retrieved, matured, fertilized and cleaved oocytes showed highly significant difference (P≤ 0.0001) when compared among different age groups. There was no significant difference in average morphokinetic time-points among young versus old women.Whereas timely cleaved embryos showed significant difference in tPNa i.e. time for pronuclear appearance (P≤ 0.001), t4 and t5 i.e. time for 4 and 5-cell cleavage (P≤ 0.05)among different age groups. The clinical pregnancy rates showed a decline with increasing age. These results indicate the effect of female age on time-lapse embryo morphokinetic parameters. In future the addition of time-lapse analysis in routine IVF can help to improve the success rate by selecting the most viable embryos for uterine transfer.

Keywords

Human embryo Time-lapse imaging Female age Blastocyst Early cleavage Morphokinetics Implantation.
Akhter, N., & Shahab, M. (2017). Morphokinetic analysis of human embryo development and its relationship to the female age: a retrospective time-lapse imaging study. Cellular and Molecular Biology, 63(8), 84–92. https://doi.org/10.14715/cmb/2017.63.8.18
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References
  1. Mio Y. Morphological analysis of human embryonic development using time-lapse cinematography. JMamm Ova Res. 2006;23:27–35.
  2. Ebner T, Yaman C, Moser M, Sommergruber M, Feichtinger O, Tews G. Prognostic value of first polar body morphology on fertilization rate and embryo quality in intracytoplasmic sperm injection. Hum Reprod. 2000;15(2):427–30.
  3. Scott LA, Smith S. The successful use of pronuclear embryo transfers the day following oocyte retrieval. Hum Reprod. 1998;13(4):1003–13.
  4. Van Montfoort AP, Dumoulin JC, Kesters AD, Evers JL. Early cleavage is a valuable addition to existing embryo selection parameters: a study using single embryo transfers. Hum Reprod. 2004;19(9):2103–8.
  5. Chamayou S, Patrizio P, Storaci G, Tomaselli V, Alecci C, Ragolia C, Crescenzo C, Guglielmino A: The use of morphokinetic parameters to select all embryos with full capacity to implant. J Assist Reprod Genet 2013, 30:703–710.
  6. Conaghan J, Chen AA, Willman SP, Ivani K, Chenette PE, Boostanfar R, Baker VL, Adamson GD, Abusief ME, Gvakharia M, Loewke KE, Shen S: Improving embryo selection using a computer-automated time-lapse image analysis test plus day 3 morphology: results from a prospective multicenter trial. Fertil Steril 2013, 100:412–419. e415.
  7. Cruz M, Gadea B, Garrido N, Pedersen KS, Martinez M, Perez-Cano I, Munoz M, Meseguer M: Embryo quality, blastocyst and ongoing pregnancy rates in oocyte donation patients whose embryos were monitored by time-lapse imaging. J Assist Reprod Genet 2011, 28:569–573.
  8. Dal Canto M, Coticchio G, Mignini Renzini M, De Ponti E, Novara PV, Brambillasca F, Comi R, Fadini R: Cleavage kinetics analysis of human embryos predicts development to blastocyst and implantation. Reprod Biomed Online 2012, 25:474–480.
  9. Kirkegaard K, Kesmodel US, Hindkjaer JJ, Ingerslev HJ: Time-lapse parameters as predictors of blastocyst development and pregnancy outcome in embryos from good prognosis patients: a prospective cohort study. Hum Reprod 2013, 28:2643–2651.
  10. Meseguer M, Herrero J, Tejera A, Hilligsoe KM, Ramsing NB, Remohi J: The use of morphokinetics as a predictor of embryo implantation. Hum Reprod 2011, 26:2658–2671.
  11. Wong CC, Loewke KE, Bossert NL, Behr B, De Jonge CJ, Baer TM, Reijo Pera RA: Non-invasive imaging of human embryos before embryonic genome activation predicts development to the blastocyst stage. Nat Biotechnol 2010, 28:1115–1121.
  12. Kirkegaard K, Hindkjaer JJ, Ingerslev HJ. Human embryonic development after blastomere removal: a time-lapse analysis. Hum Reprod 2012;27:97–105.
  13. Kirkegaard K, Agerholm IE, Ingerslev HJ. Time-lapse monitoring as a tool for clinical embryo assessment. Hum Reprod 2012;27:1277–85.
  14. Ciray HN, Aksoy T, Goktas C, Ozturk B, Bahceci M. Time-lapse evaluation of human embryo development in single versus sequential culture media – a sibling oocyte study. J Assist Reprod Genet 2012;29:891–900.
  15. Muñoz M, Cruz M, Humaidan P, Garrido N, Pérez-Cano I, Meseguer M. The type of GnRH analogue used during controlled ovarian stimulation influences early embryo developmental kinetics: a time-lapse study. Eur J Obstet Gynecol Reprod Biol 2013;168:167–172.
  16. Lemmen JG, Agerholm I, Ziebe S. Kinetic markers of human embryo quality using time-lapse recordings of IVF/ICSI-fertilized oocytes. Reprod Biomed Online 2008;17:385–391.
  17. Bellver J, Mifsud A, Grau N, Privitera L, Meseguer M. Similar morphokinetic patterns in embryos derived from obese and normoweight infertile women: a time-lapse study. Hum Reprod 2013;28:794–800.
  18. Fréour T, Dessolle L, Lammers J, Lattes S, Barrière P. Comparison of embryo morphokinetics after in vitro fertilization-intracytoplasmic sperm injection in smoking and nonsmoking women. Fertil Steril 2013; 99:1944–1950.
  19. Kirkegaard K, Hindkjaer JJ, Ingerslev HJ. Effect of oxygen concentration on human embryo development evaluated by time-lapse monitoring. Fertil Steril 2013a;99:738–744.
  20. Ahlström A, Westin C, Reismer E, Wikland M, Hardarson T. Trophectoderm morphology: an important parameter for predicting live birth after single balstocyst transfer. Hum Reprod. 2011; 26: 3289-3296.
  21. Ahlström A, Westin C, Wikland M, Hardarson T. prediction of live birth in frozen-thawed single blastocyst transfer cycles by pre-freeze and post-thaw morphology. Hum Reprod. 2013; 28: 1199-1209.
  22. Bos-Mikich A, Mattos ALG, Ferrari AN. Early cleavage of human embryos: an effective method for predicting IVF/ ICSI outcome. Hum Reprod. 2001; 16: 2658-2661.
  23. Van Kooij RJ, Looman CW, Habbema JD, Dorland M, Velde ER. Age-dependent decrease in embryo implantation rate after in vitro fertilization. 1996; Fertil Steril. 66; 769-775.
  24. Lass A, Croucher C, Duffy S, Dawson K, Margara R, Winston RM. One thousand initiated cycles of in vitro fertilization in women > or = 40 years of age. Fertil Steril. 1998; 70: 1030-1034.
  25. Schoolcraft WB, Gardner DK, Lane M, Schlenker T, Hamilton F. Blastocyst culture and transfer: analysis of results and parameters affecting outcome in two in vitro fertilization programs. Fertil Steril. 1999; 72: 604-609.
  26. Schoolcraft L, Alvero R, Leondires M, Miller B. The morphology of human pronuclear embryos is positively related to blastocyst development and implantation. Hum Reprod. 2000; 15, 2394-2403.
  27. Wittemer C, Bettahar-LeLebugle K, Ohl J, Rongiéres C, Nisand I, Gerlinger P. Zygote evaluation: an efficient tool for embryo selection. Hum Reprod. 2000; 15; 2591-2597.
  28. Gardner DK, Schoocraft WB. In vitro culture of human blastocysts: In: Janson R, Mortimer D (eds). Towards Reproductive certainty: Infertility and Genetics beyond 1999. Camforth; Parthenon Press 199.
  29. Desai NN, Goldstein J, Rowland DY, Goldfarb JM: Morphological evaluation of human embryos and derivation of an embryo quality scoring system specific for day 3 embryos: a preliminary study. Hum Reprod 2000, 15:2190–2196.
  30. Machtinger R, Racowsky C: Morphological systems of human embryo assessment and clinical evidence. Reprod Biomed Online 2013, 26:210–221.
  31. Desai N, Kinzer D, Loeb A, Goldfarb J: Use of Synthetic Serum Substitute and alpha-minimum essential medium for the extended culture of human embryos to the blastocyst stage. Hum Reprod 1997, 12:328–335.
  32. Malizia BA, Hacker MR, Penzias AS. Cumulative live-birth rates after in vitro fertilization. N Engl J Med 2009;360:236-43.
  33. Marcus SF, Brinsden PR. In-vitro fertilization and embryo transfer in women aged 40 years and over. Hum Reprod Update 1996;2: 459-68.
  34. Dain L, Auslander R, Dirnfeld M. The effect of paternal age on as¬sisted reproduction outcome. Fertil Steril 2011;95:1-8.
  35. Dew JE, Don RA, Hughes GJ, Johnson TC, Steigrad SJ. The influ¬ence of advanced age on the outcome of assisted reproduction. J Assist Reprod Genet 1998;15:210-4.
  36. Ziebe S, Loft A, Petersen JH, Andersen AG, Lindenberg S, Peters¬en K, et al. Embryo quality and developmental potential is com¬promised by age. Acta Obstet Gynecol Scand 2001;80:169-74.
  37. Szamatowicz M, Grochowski D. Fertility and infertility in aging women. Gynecol Endocrinol 1998;12:407-13.
  38. Pantos K, Athanasiou V, Stefanidis K, Stavrou D, Vaxevanoglou T, Chronopoulou M. Influence of advanced age on the blastocyst development rate and pregnancy rate in assisted reproductive technology. Fertil Steril 1999;71:1144-6.
  39. Ventura SJ, Mosher WD, Curtin SC, Abma JC, Henshaw S. Trends in pregnancies and pregnancy rates by outcome: estimates for the United States, 1976-96. Vital Health Stat 21 2000;(56):1-47.
  40. Ron-El R, Raziel A, Strassburger D, Schachter M, Kasterstein E, Friedler S. Outcome of assisted reproductive technology in wom¬en over the age of 41. Fertil Steril 2000;74:471-5.
  41. Devroey P, Godoy H, Smitz J, Camus M, Tournaye H, Derde MP, et al. Female age predicts embryonic implantation after ICSI: a case-controlled study. Hum Reprod 1996;11:1324-7.
  42. Dor J, Seidman DS, Ben-Shlomo I, Levran D, Ben-Rafael Z, Mashiach S. Cumulative pregnancy rate following in-vitro fertil¬ization: the significance of age and infertility aetiology. Hum Re¬prod 1996;11:425-8.
  43. Widra EA, Gindoff PR, Smotrich DB, Stillman RJ. Achieving multi¬ple-order embryo transfer identifies women over 40 years of age with improved in vitro fertilization outcome. Fertil Steril 1996;65:103-8.
  44. Maheshwari A, Hamilton M, Bhattacharya S. Effect of female age on the diagnostic categories of infertility. Hum Reprod 2008;23: 538-42.
  45. Wood C, Calderon I, Crombie A. Age and fertility: results of assist¬ed reproductive technology in women over 40 years. J Assist Re¬prod Genet 1992;9:482-4.
  46. Abdalla HI, Burton G, Kirkland A, Johnson MR, Leonard T, Brooks AA, et al. Age, pregnancy and miscarriage: uterine versus ovari¬an factors. Hum Reprod 1993;8:1512-7.
  47. De Croo I, Van der Elst J, Everaert K, De Sutter P, Dhont M. Fertil¬ization, pregnancy and embryo implantation rates after ICSI in cases of obstructive and non-obstructive azoospermia. Hum Re¬prod 2000;15:1383-8.
  48. Schieve LA, Peterson HB, Meikle SF, Jeng G, Danel I, Burnett NM, et al. Live-birth rates and multiple-birth risk using in vitro fertil¬ization. JAMA 1999;282:1832-8.
  49. Silber SJ, Nagy Z, Devroey P, Camus M, Van Steirteghem AC. The effect of female age and ovarian reserve on pregnancy rate in male infertility: treatment of azoospermia with sperm retrieval and intracytoplasmic sperm injection. Hum Reprod 1997;12: 2693-700.
  50. Lee SH, Song H, Park YS, Koong MK, Song IO, Jun JH. Poor sperm quality affects clinical outcomes of intracytoplasmic sperm in¬jection in fresh and subsequent frozen-thawed cycles: potential paternal effects on pregnancy outcomes. Fertil Steril 2009;91: 798-804.
  51. Munne S, Alikani M, Tomkin G, Grifo J, Cohen J. Embryo mor¬phology, developmental rates, and maternal age are correlated with chromosome abnormalities. Fertil Steril 1995;64:382-91.
  52. Martin RH. Meiotic errors in human oogenesis and spermato¬genesis. Reprod Biomed Online 2008;16:523-31.
  53. Flamigni C. Egg donation to women over 40 years of age. Hum Reprod 1993;8:1343-4.
  54. Borini A, Bianchi L, Violini F, Maccolini A, Cattoli M, Flamigni C. Oocyte donation program: pregnancy and implantation rates in women of different ages sharing oocytes from single donor. Fer¬til Steril 1996;65:94-7.
  55. Ramsey JE, Greer I, Sattar N. Obesity and reproduction. BMJ 2006;333: 1159–62.
  56. Comstock IA, Kim S, Behr B, Lathi RB. Increased body mass index negatively impacts blastocyst formation rate in normal responders undergoing in vitro fertilization. J Assist Reprod Genet 2015;32:1299–304
  57. Yan J, Wu K, Tang R, et al. Effect of maternal age on the outcomes of in vitro fertilization and embryo transfer (IVF-ET). Science China Life Sci 2012;55:694–8.
  58. Preutthipan S, Amso N, Curtis P, Shaw RW. Effect of maternal age on clinical outcome in women undergoing in vitro fertilization and embryo transfer (IVF-ET). J Med Assoc Thailand¼Chotmaihet Thangphaet 1996;79:347–52.
  59. Eichenlaub-Ritter U. Genetics of oocyte ageing. Maturitas 1998;30: 143–69.
  60. Fragouli E, Wells D, Delhanty JDA. Chromosome abnormalities in the human oocyte. Cytogenet Genome Res 2011;133:107–18.
  61. Eichenlaub-Ritter U, Vogt E, Yin H, Gosden R. Spindles, mitochondria and redox potential in ageing oocytes. Reprod Biomed Online 2004;8:45–58.
  62. Akarsu S., Gode F., Isik A. Z., Celenk H., Tamer F. B. and Erkilinc S. Comparison of the morphokinetic parameters of embryos according to ovarian reserve in IVF cycles. Gynecol Endocrinol. 2017, 17: 1-4.
  63. Hlinka D1, Kać¾atová B, Uhrinová I, Dolinská S, Rutarová J, Rezáćová J, Lazarovská S, Dudáš M. Time-lapse cleavage rating predicts human embryo viability. Physiol Res. 2012;61(5):513-25.
  64. Herrero J, Tejera A, Albert C, Vidal C, de Los Santos MJ, Meseguer M: A time to look back: analysis of morphokinetic characteristics of human embryo development. Fertil Steril. 2013, 100: 1602-1609
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Author Biography

N. Akhter, Department of Animal Sciences, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan
Department of Animal Sciences, PhD Scholar
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