The influence of preincubation time of prepared sperm before IVF on fertilization, embryo developmental competence and the reproductive outcomes
Abstract
Objectives: It has been provided that if incubation time of prepared sperm can affect sperm motility and DNA fragment, but little is known about the influence of sperm preincubation time (SI) on the sperm’s fertilizing ability, subsequent embryonic development and pregnancy outcomes in in vitro fertilization (IVF). The aim of this study was to explore the association of SI with fertilization rate, embryo development and clinical outcomes in IVF, further, to find an optimal preincubation time for prepared sperm before insemination in IVF.
Material and methods: This retrospective cohort study included a total of 1453 infertile couples undergoing IVF in our center performed from January 2016 to January 2019. Sperm were preincubated at 37℃ 6% CO2 for different times before insemination. Preincubation time associated with fertilization rate (FR), 2PN rate, D3 good quality embryo rate, fresh embryo implantation rate (IR), blastocyst formation rate, cumulative pregnancy rate (CPR), cumulative ongoing pregnancy rate (COPR), cumulative live birth rate (CLBR), newborn health and gender ratio were analyzed by chi-square analysis.
Results: FR and 2PN rate of SI more than four hours SI groups (> 4 h SI group) decreased significantly compared with other SI groups (p < 0.01). There were no significant differences of the D3 high quality embryo rate among five SI groups. The blastocyst formation rate of > 4 h SI group was significantly lower than that of 2–3 h SI group (45.5% vs 56.1%, p < 0.05); and 1–2 h SI group also had significant difference with 2–3 h and 3–4 h SI group (48.9% vs 56.1% and 54.6%, p < 0.05). There were a significant decrease of fresh IR and CPR in ≤1 h SI group compared with 1–2 h SI group (19.6% vs. 38.0%, p < 0.05; 62.7% vs 73.7%, p < 0.05); ≤ 1 h SI group also have the lowest CLBR (45.6%), it had statistic differences with 1–2 SI group and 3–4 SI group (45.6% vs 63.2%, p < 0.01; 45.6% vs 61.2%, p < 0.05).
Conclusions: The sperm preincubated time at 37℃ 6% CO2 before insemination could influence sperm fertilizing ability, blastocyst formation, embryo implantation and CLBR in IVF cycles. The best time for prepared sperm preincubation at 37℃ is one to four hours before insemination in IVF.
Keywords: spermpreincubation timeIVFfertilization rateembryo developmentcumulative live birth
References
- Whitaker M, Fraser LR, Weir BJ. Cell messengers at fertilization. . J Reprod Fertil. 1990; 42(Suppl): 1–254.
- Guraya S. Cellular and molecular biology of capacitation and acrosome reaction in spermatozoa. International Review of Cytology. 2000; 199: 1–64.
- Marı́n-Briggiler C, Tezón J, Miranda P, et al. Effect of incubating human sperm at room temperature on capacitation-related events. Fertility and Sterility. 2002; 77(2): 252–259.
- Jeulin C, Serres C, Jouannet P. The effects of centrifugation, various synthetic media and temperature on the motility and vitality of human spermatozoa. Reprod Nutr Dev. 1982; 22(1A): 81–91.
- Lachaud C, Tesarik J, Cañadas ML, et al. Apoptosis and necrosis in human ejaculated spermatozoa. Hum Reprod. 2004; 19(3): 607–610.
- Scott L, Smith S. Mouse in vitro fertilization, embryo development and viability, and human sperm motility in substances used for human sperm preparation for assisted reproduction. Fertil Steril. 1997; 67(2): 372–381.
- Calamera JC, Fernandez PJ, Buffone MG, et al. Effects of long-term in vitro incubation of human spermatozoa: functional parameters and catalase effect. Andrologia. 2001; 33(2): 79–86.
- Nabi A, Khalili MA, Halvaei I, et al. Prolonged incubation of processed human spermatozoa will increase DNA fragmentation. Andrologia. 2014; 46(4): 374–379.
- Zhang XD, Chen MY, Gao Y, et al. The effects of different sperm preparation methods and incubation time on the sperm DNA fragmentation. Hum Fertil (Camb). 2011; 14(3): 187–191.
- Mallett PJ, Stock CE, Fraser LR. Acrosome loss in human sperm incubated in vitro under capacitating conditions. Int J Androl. 1985; 8(5): 357–364.
- Dolci S, Manna C, Jannini EA, et al. ATP-content and kinetics of acrosome reaction in human spermatozoa: influence of various culture media and incubation time. Andrologia. 1988; 20(2): 169–172.
- WHO laboratory manual for the examination and processing of human semen. 5th ed. Geneva. World Health Organization press. 2010.
- ALPHA Scientists In Reproductive Medicine, ESHRE Special Interest Group Embryology. Istanbul consensus workshop on embryo assessment: proceedings of an expert meeting. Reprod Biomed Online. 2011; 22(6): 632–646.
- Gardner DK, Schoolcraft WB. In vitro culture of the human blastocyst. . In: Jansen R, Mortimer D. ed. Towards Reproductive Certainty: Infertility and gGenetics Beyond. Parthenon Publishing, Carnforth 1999: 378–388.
- Zollner U, Martin S, Liebermann J, et al. Evaluation of a cut-off value for sperm motility after different hours of incubation to select the suitable reproductive technology (IVF or ICSI). Acta Obstet Gynecol Scand. 1999; 78(4): 326–331.
- Oleszczuk K, Giwercman A, Bungum M. Sperm chromatin structure assay in prediction of in vitro fertilization outcome. Andrology. 2016; 4(2): 290–296.
- Gosálvez J, Cortés-Gutierez E, López-Fernández C, et al. Sperm deoxyribonucleic acid fragmentation dynamics in fertile donors. Fertil Steril. 2009; 92(1): 170–173.
- de Lamirande E, Gagnon C. Capacitation-associated production of superoxide anion by human spermatozoa. Free Radic Biol Med. 1995; 18(3): 487–495.
- Lefièvre L, Jha KN, de Lamirande E, et al. Activation of protein kinase A during human sperm capacitation and acrosome reaction. J Androl. 2002; 23(5): 709–716.
- Harrison RAP. Rapid PKA-catalysed phosphorylation of boar sperm proteins induced by the capacitating agent bicarbonate. Mol Reprod Dev. 2004; 67(3): 337–352.
- O’Flaherty C, de Lamirande E, Gagnon C. Phosphorylation of the Arginine‑X‑X‑(Serine/ Threonine) motile in human sperm proteins during capacitation: modulation and protein kinase A dependency. Mol Hum Reprod. 2004; 10: 355–63.
- Leclerc P, de Lamirande E, Gagnon C. Regulation of protein-tyrosine phosphorylation and human sperm capacitation by reactive oxygen derivatives. Free Radic Biol Med. 1997; 22(4): 643–656.
- de Lamirande E, Gagnon C. Paradoxical effect of reagents for sulfhydryl and disulfide groups on human sperm capacitation and superoxide production. Free Radic Biol Med. 1998; 25(7): 803–817.
- de Lamirande E, Gagnon C. Redox control of changes in protein sulfhydryl levels during human sperm capacitation. Free Radic Biol Med. 2003; 35(10): 1271–1285.
- Xue LT, Wang RX, He B, et al. Effect of sperm DNA fragmentation on clinical outcomes for Chinese couples undergoing in vitro fertilization or intracytoplasmic sperm injection. J Int Med Res. 2016; 44(6): 1283–1291.
- Simon L, Murphy K, Shamsi MB, et al. Paternal influence of sperm DNA integrity on early embryonic development. Hum Reprod. 2014; 29(11): 2402–2412.
- Breznik BP, Kovačič BSc, Veljko Vlaisavljević V. Are sperm DNA fragmentation, hyperactivation, and hyaluronan- binding ability predictive for fertilization and embryo development in in vitro fertilization and intracytoplasmic sperm injection? Fertil Steril. 2013; 99(5): 1233–1241.
- Borges Jr E, Zanetti BF, Setti AS, et al. Sperm DNA fragmentation is correlated with poor embryo development, lower implantation rate, and higher miscarriage rate in reproductive cycles of non-male factor infertility. Ferti Steril. 2019; 112(3): 483–490.
- Zhao J, Zhang Q, Wang YG, et al. Whether sperm deoxyribonucleic acid fragmentation has an effect on pregnancy and miscarriage after in vitro fertilization /intracytoplasmic sperm injection: a systematic review and meta-analysis. Fertil Steril. 2014; 102: 998–1005e8.
