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Chapter 10 - Examining the Safety of ICSI Using Animal Models

Published online by Cambridge University Press:  02 December 2021

Gianpiero D. Palermo
Cornell Institute of Reproductive Medicine, New York
Zsolt Peter Nagy
Reproductive Biology Associates, Atlanta, GA
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There is no doubt that intracytoplasmic sperm injection (ICSI) has been a major breakthrough in treating male infertility, accounting for 70–80% of all the cycles performed worldwide. Surprisingly, there were very few animal studies conducted before the first babies were born, in part because of technical challenges that were experienced in most animal systems. Technological advancements were required to develop appropriate animal models for assessing the safety of ICSI. These studies identified many cytoskeletal changes that occurred in the oocyte cytoplasm after ICSI versus natural conception or even IVF, raising concerns about the long-term health of ICSI offspring. This chapter summarizes the animal models that have contributed to our understanding of the cellular and molecular aspects of ICSI, as well as providing models to investigate both the developmental origins of adult disease and transgenerational implications as they relate to ICSI.

Manual of Intracytoplasmic Sperm Injection in Human Assisted Reproduction
With Other Advanced Micromanipulation Techniques to Edit the Genetic and Cytoplasmic Content of the Oocyte
, pp. 95 - 102
Publisher: Cambridge University Press
Print publication year: 2021

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Palermo, G., et al., Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte. Lancet, 1992. 340(8810): pp. 17–8.CrossRefGoogle ScholarPubMed
Van Steirteghem, A.C., et al., Higher success rate by intracytoplasmic sperm injection than by subzonal insemination. Report of a second series of 300 consecutive treatment cycles. Hum Reprod, 1993. 8(7): pp. 1055–60.CrossRefGoogle ScholarPubMed
Fauser, B.C.J.M. Towards the global coverage of a unified registry of IVF outcomes. Reprod Biomed Online, 2019. 38: pp. 133–7.CrossRefGoogle ScholarPubMed
Nyboe Andersen, A., Carlsen, E., and Loft, A., Trends in the use of intracytoplasmatic sperm injection marked variability between countries. Hum Reprod Update, 2008. 14(6): pp. 593604.CrossRefGoogle ScholarPubMed
Vernaeve, V., et al., Pregnancy outcome and neonatal data of children born after ICSI using testicular sperm in obstructive and non-obstructive azoospermia. Hum Reprod, 2003. 18(10): pp. 2093–7.CrossRefGoogle ScholarPubMed
Palermo, G.D., Neri, Q.V., and Rosenwaks, Z., To ICSI or not to ICSI. Semin Reprod Med, 2015. 33(2): pp. 92102.Google ScholarPubMed
Merchant, R., Gandhi, G., and Allahbadia, G.N., In vitro fertilization/intracytoplasmic sperm injection for male infertility. Indian J Urol, 2011. 27(1): pp. 121–32.Google ScholarPubMed
O’Neill, C.L., et al., Development of ICSI. Reproduction, 2018. 156(1): pp. F51F58.CrossRefGoogle ScholarPubMed
Schatten, G., et al., Cell and molecular biological challenges of ICSI: ART before science? J Law Med Ethics, 1998. 26(1): pp. 2937.CrossRefGoogle ScholarPubMed
Salamone, D.F., Canel, N.G., and Rodriguez, M.B., Intracytoplasmic sperm injection in domestic and wild mammals. Reproduction, 2017. 154(6): pp. F111F124.CrossRefGoogle ScholarPubMed
Hosoi, Y. and Iritani, A., Rabbit microfertilization. Mol Reprod Dev, 1993. 36(2): pp. 282–4.CrossRefGoogle ScholarPubMed
Kimura, Y. and Yanagimachi, R., Intracytoplasmic sperm injection in the mouse. Biol Reprod, 1995. 52(4): pp. 709–20.CrossRefGoogle ScholarPubMed
Hewitson, L.C., et al., Microtubule and chromatin configurations during rhesus intracytoplasmic sperm injection: successes and failures. Biol Reprod, 1996. 55(2): pp. 271–80.CrossRefGoogle ScholarPubMed
Dozortsev, D., et al., Intracytoplasmic sperm injection in the rat. Zygote, 1998. 6(2): pp. 143–7.CrossRefGoogle ScholarPubMed
Gomez, M.C., et al., Sheep oocyte activation after intracytoplasmic sperm injection (ICSI). Reprod Fertil Dev, 1998. 10(2): pp. 197205.CrossRefGoogle Scholar
Pope, C.E., et al., Development of embryos produced by intracytoplasmic sperm injection of cat oocytes. Anim Reprod Sci, 1998. 53(1–4): pp. 221–36.CrossRefGoogle ScholarPubMed
Kolbe, T. and Holtz, W., Intracytoplasmic injection (ICSI) of in vivo or in vitro matured oocytes with fresh ejaculated or frozen-thawed epididymal spermatozoa and additional calcium-ionophore activation in the pig. Theriogenology, 1999. 52(4): pp. 671–82.CrossRefGoogle ScholarPubMed
Cochran, R., et al., Production of live foals from sperm-injected oocytes harvested from pregnant mares. J Reprod Fertil Suppl, 2000(56): pp. 503–12.Google Scholar
Yamauchi, Y., Yanagimachi, R., and Horiuchi, T., Full-term development of golden hamster oocytes following intracytoplasmic sperm head injection. Biol Reprod, 2002. 67(2): pp. 534–9.CrossRefGoogle ScholarPubMed
Yanagimachi, R., Intracytoplasmic sperm injection experiments using the mouse as a model. Hum Reprod, 1998. 13 Suppl 1: pp. 8798.CrossRefGoogle ScholarPubMed
Singh Parmer, M., et al., Intracytoplasmic sperm injection (ICSI) and its applications in veterinary sciences: An overview. Sci Int, 2013. 1(8): pp. 266270.Google Scholar
Oikawa, T., et al., Evaluation of activation treatments for blastocyst production and birth of viable calves following bovine intracytoplasmic sperm injection. Anim Reprod Sci, 2005. 86(3–4): pp. 187–94.CrossRefGoogle ScholarPubMed
Garcia-Rosello, E., et al., Intracytoplasmic sperm injection in livestock species: an update. Reprod Domest Anim, 2009. 44(1): pp. 143–51.CrossRefGoogle ScholarPubMed
Li, M.W., et al., Long-term storage of mouse spermatozoa after evaporative drying. Reproduction, 2007. 133(5): pp. 919–29.CrossRefGoogle ScholarPubMed
Li, M.W., et al., Damage to chromosomes and DNA of rhesus monkey sperm following cryopreservation. J Androl, 2007. 28(4): pp. 493501.CrossRefGoogle ScholarPubMed
Ramalho-Santos, J., et al., ICSI choreography: fate of sperm structures after monospermic rhesus ICSI and first cell cycle implications. Hum Reprod, 2000. 15(12): pp. 2610–20.CrossRefGoogle ScholarPubMed
Sutovsky, P., et al., Intracytoplasmic sperm injection for Rhesus monkey fertilization results in unusual chromatin, cytoskeletal, and membrane events, but eventually leads to pronuclear development and sperm aster assembly. Hum Reprod, 1996. 11(8): pp. 1703–12.CrossRefGoogle ScholarPubMed
Ramalho-Santos, J., et al., SNAREs in mammalian sperm: possible implications for fertilization. Dev Biol, 2000. 223(1): pp. 5469.CrossRefGoogle ScholarPubMed
Hewitson, L., et al., Unique checkpoints during the first cell cycle of fertilization after intracytoplasmic sperm injection in rhesus monkeys. Nat Med, 1999. 5(4): pp. 431–3.CrossRefGoogle ScholarPubMed
Motoishi, M., et al., Examination of the safety of intracytoplasmic injection procedures by using bovine zygotes. Hum Reprod, 1996. 11(3): pp. 618–20.CrossRefGoogle ScholarPubMed
Yanagimachi, R., Intracytoplasmic injection of spermatozoa and spermatogenic cells: its biology and applications in humans and animals. Reprod Biomed Online, 2005. 10(2): pp. 247–88.CrossRefGoogle Scholar
Mansour, R., Intracytoplasmic sperm injection: a state of the art technique. Hum Reprod Update, 1998. 4(1): pp. 4356.CrossRefGoogle ScholarPubMed
Rubino, P., et al., The ICSI procedure from past to future: a systematic review of the more controversial aspects. Hum Reprod Update, 2016. 22(2): pp. 194227.Google ScholarPubMed
Asch, R., et al., The stages at which human fertilization arrests: microtubule and chromosome configurations in inseminated oocytes which failed to complete fertilization and development in humans. Hum Reprod, 1995. 10(7): pp. 1897–906.CrossRefGoogle ScholarPubMed
Simerly, C.R., et al., Fertilization and cleavage axes differ in primates conceived by conventional (IVF) versus intracytoplasmic sperm injection (ICSI). Sci Rep, 2019. 9(1): p. 15282.CrossRefGoogle Scholar
Bras, M, et al., The use of a mouse zygote quality control system for training purposes and toxicity determination in an ICSI programme. Hum Reprod, 1994. 9: pp. 2324.Google Scholar
Strehler, E., et al., Detrimental effects of polyvinylpyrrolidone on the ultrastructure of spermatozoa (Notulae seminologicae 13). Hum Reprod, 1998. 13(1): pp. 120–3.CrossRefGoogle Scholar
Kato, Y. and Nagao, Y., Effect of PVP on sperm capacitation status and embryonic development in cattle. Theriogenology, 2009. 72(5): pp. 624–35.CrossRefGoogle ScholarPubMed
Hewitson, L., et al., Fertilization and embryo development to blastocysts after intracytoplasmic sperm injection in the rhesus monkey. Hum Reprod, 1998. 13(12): pp. 3449–55.CrossRefGoogle ScholarPubMed
Hewitson, L., et al., Rhesus offspring produced by intracytoplasmic injection of testicular sperm and elongated spermatids. Fertil Steril, 2002. 77(4): pp. 794801.CrossRefGoogle ScholarPubMed
Chan, A.W., et al., Foreign DNA transmission by ICSI: injection of spermatozoa bound with exogenous DNA results in embryonic GFP expression and live rhesus monkey births. Mol Hum Reprod, 2000. 6(1): pp. 2633.CrossRefGoogle ScholarPubMed
Feuer, S.K., Camarano, L., and Rinaudo, P.F., ART and health: clinical outcomes and insights on molecular mechanisms from rodent studies. Mol Hum Reprod, 2013. 19(4): pp. 189204.CrossRefGoogle ScholarPubMed
Fauser, B.C., et al., Health outcomes of children born after IVF/ICSI: a review of current expert opinion and literature. Reprod Biomed Online, 2014. 28(2): pp. 162–82.CrossRefGoogle ScholarPubMed
Hansen, M., et al., Assisted reproductive technologies and the risk of birth defects–a systematic review. Hum Reprod, 2005. 20(2): pp. 328–38.CrossRefGoogle ScholarPubMed
Manipalviratn, S., DeCherney, A., and Segars, J., Imprinting disorders and assisted reproductive technology. Fertil Steril, 2009. 91(2): pp. 305–15.CrossRefGoogle ScholarPubMed
Belva, F., Bonduelle, M., and Tournaye, H., Endocrine and reproductive profile of boys and young adults conceived after ICSI. Curr Opin Obstet Gynecol, 2019. 31(3): pp. 163169.CrossRefGoogle Scholar
Sanchez-Calabuig, M.J., et al., Potential health risks associated to ICSI: insights from animal models and strategies for a safe procedure. Front Public Health, 2014. 2: p. 241.CrossRefGoogle ScholarPubMed
Vrooman, L.A. and Bartolomei, M.S., Can assisted reproductive technologies cause adult-onset disease? Evidence from human and mouse. Reprod Toxicol, 2017. 68: pp. 7284.CrossRefGoogle ScholarPubMed
Yamagata, K., Suetsugu, R., and Wakayama, T., Assessment of chromosomal integrity using a novel live-cell imaging technique in mouse embryos produced by intracytoplasmic sperm injection. Hum Reprod, 2009. 24(10): pp. 2490–9.CrossRefGoogle ScholarPubMed
Barker, D.J., The developmental origins of adult disease. J Am Coll Nutr, 2004. 23(6 Suppl): pp. 588S595S.CrossRefGoogle ScholarPubMed
Scott, K.A., et al., Glucose parameters are altered in mouse offspring produced by assisted reproductive technologies and somatic cell nuclear transfer. Biol Reprod, 2010. 83(2): pp. 220–7.CrossRefGoogle ScholarPubMed
Rexhaj, E., et al., Mice generated by in vitro fertilization exhibit vascular dysfunction and shortened life span. J Clin Invest, 2013. 123(12): pp. 5052–60.CrossRefGoogle ScholarPubMed
Yu, Y., et al., Microinjection manipulation resulted in the increased apoptosis of spermatocytes in testes from intracytoplasmic sperm injection (ICSI) derived mice. PLoS One, 2011. 6(7): p. e22172.CrossRefGoogle ScholarPubMed
Fernandez-Gonzalez, R., et al., Long-term effects of mouse intracytoplasmic sperm injection with DNA-fragmented sperm on health and behavior of adult offspring. Biol Reprod, 2008. 78(4): pp. 761–72.CrossRefGoogle ScholarPubMed
Schatten, G., The centrosome and its mode of inheritance: the reduction of the centrosome during gametogenesis and its restoration during fertilization. Dev Biol, 1994. 165(2): pp. 299335.CrossRefGoogle ScholarPubMed
Schatten, G., Simerly, C., and Schatten, H., Microtubule configurations during fertilization, mitosis, and early development in the mouse and the requirement for egg microtubule-mediated motility during mammalian fertilization. Proc Natl Acad Sci U S A, 1985. 82(12): pp. 4152–6.CrossRefGoogle ScholarPubMed
Simerly, CR, et al., Male infertility as a result of disorders in the paternally inherited human centrosome. South African J Sci 1997. 92: pp. 548557.Google Scholar
Terada, Y., et al., Human sperm aster formation after intracytoplasmic sperm injection with rabbit and bovine eggs. Fertil Steril, 2002. 77(6): pp. 1283–4.CrossRefGoogle ScholarPubMed
Terada, Y., Human sperm centrosomal function during fertilization, a novel assessment for male sterility. Hum Cell, 2004. 17(4): pp. 181–6.Google ScholarPubMed
Terada, Y., et al., Use of Mammalian eggs for assessment of human sperm function: molecular and cellular analyses of fertilization by intracytoplasmic sperm injection. Am J Reprod Immunol, 2004. 51(4): pp. 290–3.CrossRefGoogle ScholarPubMed
Terada, Y., et al., Atypical decondensation of the sperm nucleus, delayed replication of the male genome, and sex chromosome positioning following intracytoplasmic human sperm injection (ICSI) into golden hamster eggs: does ICSI itself introduce chromosomal anomalies? Fertil Steril, 2000. 74(3): pp. 454–60.CrossRefGoogle ScholarPubMed
Luetjens, C.M., Payne, C., and Schatten, G., Non-random chromosome positioning in human sperm and sex chromosome anomalies following intracytoplasmic sperm injection. Lancet, 1999. 353(9160): p. 1240.CrossRefGoogle ScholarPubMed
Zalenskaya, I.A. and Zalensky, A.O., Non-random positioning of chromosomes in human sperm nuclei. Chromosome Res, 2004. 12(2): pp. 163–73.CrossRefGoogle ScholarPubMed
Feichtinger, W., Obruca, A., and Brunner, M., Sex chromosomal abnormalities and intracytoplasmic sperm injection. Lancet, 1995. 346(8989): p. 1566.CrossRefGoogle ScholarPubMed
Kasai, T., Hoshi, K., and Yanagimachi, R., Effect of sperm immobilisation and demembranation on the oocyte activation rate in the mouse. Zygote, 1999. 7(3): pp. 187–93.CrossRefGoogle ScholarPubMed
Jones, E.L., Mudrak, O., and Zalensky, A.O., Kinetics of human male pronuclear development in a heterologous ICSI model. J Assist Reprod Genet, 2010. 27(6): pp. 277–83.Google Scholar
Heindryckx, B., et al., Treatment option for sperm- or oocyte-related fertilization failure: assisted oocyte activation following diagnostic heterologous ICSI. Hum Reprod, 2005. 20(8): pp. 2237–41.CrossRefGoogle ScholarPubMed
Vanden Meerschaut, F., et al., Diagnostic and prognostic value of calcium oscillatory pattern analysis for patients with ICSI fertilization failure. Hum Reprod, 2013. 28(1): pp. 8798.CrossRefGoogle ScholarPubMed
Vanden Meerschaut, F., et al., Assisted oocyte activation following ICSI fertilization failure. Reprod Biomed Online, 2014. 28(5): p. 560–71.CrossRefGoogle ScholarPubMed
Swann, K. and Lai, F.A., PLCzeta and the initiation of Ca(2+) oscillations in fertilizing mammalian eggs. Cell Calcium, 2013. 53(1): pp. 5562.CrossRefGoogle ScholarPubMed
Vanden Meerschaut, F., et al., Assisted oocyte activation is not beneficial for all patients with a suspected oocyte-related activation deficiency. Hum Reprod, 2012. 27(7): pp. 1977–84.CrossRefGoogle Scholar
Amdani, S.N., et al., Sperm factors and oocyte activation: current controversies and considerations. Biol Reprod, 2015. 93(2): p. 50.CrossRefGoogle ScholarPubMed
Nomikos, M., et al., Phospholipase Czeta rescues failed oocyte activation in a prototype of male factor infertility. Fertil Steril, 2013. 99(1): pp. 7685.CrossRefGoogle Scholar
Yoon, S.Y., et al., Human sperm devoid of PLC, zeta 1 fail to induce Ca(2+) release and are unable to initiate the first step of embryo development. J Clin Invest, 2008. 118(11): pp. 3671–81.CrossRefGoogle ScholarPubMed
Nomikos, M., Swann, K., and Lai, F.A., Starting a new life: sperm PLC-zeta mobilizes the Ca2+ signal that induces egg activation and embryo development: an essential phospholipase C with implications for male infertility. Bioessays, 2012. 34(2): pp. 126–34.CrossRefGoogle ScholarPubMed
Sanusi, R., et al., Rescue of failed oocyte activation after ICSI in a mouse model of male factor infertility by recombinant phospholipase Czeta. Mol Hum Reprod, 2015. 21(10): pp. 783–91.CrossRefGoogle Scholar
Taylor, S.L., et al., Complete globozoospermia associated with PLCzeta deficiency treated with calcium ionophore and ICSI results in pregnancy. Reprod Biomed Online, 2010. 20(4): pp. 559–64.CrossRefGoogle ScholarPubMed
Chithiwala, Z.H., et al., Phospholipase C-zeta deficiency as a cause for repetitive oocyte fertilization failure during ovarian stimulation for in vitro fertilization with ICSI: a case report. J Assist Reprod Genet, 2015. 32(9): pp. 1415–9.CrossRefGoogle ScholarPubMed
Nazarian, H., et al., Effect of artificial oocyte activation on intra-cytoplasmic sperm injection outcomes in patients with lower percentage of sperm containing phospholipase Czeta: a randomized clinical trial. J Reprod Infertil, 2019. 20(1): pp. 39.Google ScholarPubMed
Vanden Meerschaut, F., et al., Neonatal and neurodevelopmental outcome of children aged 3–10 years born following assisted oocyte activation. Reprod Biomed Online, 2014. 28(1): pp. 5463.CrossRefGoogle ScholarPubMed
Nasr-Esfahani, M.H., Deemeh, M.R., and Tavalaee, M., Artificial oocyte activation and intracytoplasmic sperm injection. Fertil Steril, 2010. 94(2): pp. 520–6.CrossRefGoogle ScholarPubMed
van Blerkom, J., Cohen, J., and Johnson, M., A plea for caution and more research in the ‘experimental’ use of ionophores in ICSI. Reprod Biomed Online, 2015. 30(4): pp. 323–4.CrossRefGoogle ScholarPubMed
Nomikos, M., et al., Human PLCzeta exhibits superior fertilization potency over mouse PLCzeta in triggering the Ca(2+) oscillations required for mammalian oocyte activation. Mol Hum Reprod, 2014. 20(6): pp. 489–98.CrossRefGoogle ScholarPubMed

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