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Chapter 2 - Development of ICSI in Human Assisted Reproduction

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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Since the establishment of in vitro fertilization, it became quickly apparent that approximately half of the couples treated presented with a dysfunctional male gamete. To alleviate this issue, intracytoplasmic sperm injection (ICSI) was introduced to treat men with compromised semen parameters or azoospermia, and more recently high sperm chromatin fragmentation or sperm-linked oocyte activation deficiency. Because of its success, ICSI has been extended for cases with low egg yield, oocyte cryopreservation, and often for preimplantation genetic testing. Due to its versatility and reliability, ICSI has become the most popular ART and will be invaluable for emerging technologies such as in vitro gametogenesis and heritable genome editing. In this chapter, we discuss the development of ICSI, its current applications, and ongoing research that will contribute to the future of reproductive medicine.

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. 11 - 24
Publisher: Cambridge University Press
Print publication year: 2021

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Pincus, G, Enzmann, EV. Can mammalian eggs undergo normal development in vitro? Proceedings of the National Academy of Sciences of the United States of America 1934;20:121122.CrossRefGoogle ScholarPubMed
Menkin, MF, Rock, J. In vitro fertilization and cleavage of human ovarian eggs. American Journal of Obstetrics and Gynecology 1948;55:440452.CrossRefGoogle ScholarPubMed
Steptoe, PC, Edwards, RG. Birth after the reimplantation of a human embryo. Lancet 1978;2: 366.CrossRefGoogle ScholarPubMed
Palermo, G, Van Steirteghem, A. Enhancement of acrosome reaction and subzonal insemination of a single spermatozoon in mouse eggs. Molecular Reproduction and Development 1991;30:339345.CrossRefGoogle ScholarPubMed
Society for Assisted Reproductive Technology. National Summary Report, 2018, Scholar
Steptoe, PC, Edwards, RG. Reimplantation of a human embryo with subsequent tubal pregnancy. Lancet 1976;307:880882.CrossRefGoogle Scholar
Cohen, J, Edwards, R, Fehilly, C, Fishel, S, Hewitt, J, Purdy, J, Rowland, G, Steptoe, P, Webster, J. In vitro fertilization: a treatment for male infertility. Fertility and Sterility 1985;43:422432.CrossRefGoogle ScholarPubMed
Cohen, J, Malter, H, Wright, G, Kort, H, Massey, J, Mitchell, D. Partial zona dissection of human oocytes when failure of zona pellucida penetration is anticipated. Human Reproduction 1989;4:435442.CrossRefGoogle ScholarPubMed
Svalander, P, Wikland, M, Jakobsson, A-H, Forsberg, A-S. Subzonal insemination (SUZI) or in vitro fertilization (IVF) in microdroplets for the treatment of male-factor infertility. Journal of Assisted Reproduction and Genetics 1994;11:149155.CrossRefGoogle ScholarPubMed
Kiessling, AA, Loutradis, D, McShane, PM, Jackson, KV. Fertilization in trypsin-treated oocytes. Annals of the New York Academy of Sciences 1988;541:614620.CrossRefGoogle ScholarPubMed
Gordon, JW, Talansky, BE. Assisted fertilization by zona drilling: A mouse model for correction of oligospermia. Journal of Experimental Zoology 1986;239:347354.CrossRefGoogle ScholarPubMed
Tucker, MJ, Bishop, FM, Cohen, J, Wiker, SR, Wright, G. Routine application of partial zona dissection for male factor infertility. Human Reproduction 1991;6:676681.CrossRefGoogle ScholarPubMed
Ng, SC, Bongso, A, Ratnam, SS, Sathananthan, H, Chan, CL, Wong, PC, Hagglund, L, Anandakumar, C, Wong, YC, Goh, VH. Pregnancy after transfer of sperm under zona. Lancet 1988;2:790.CrossRefGoogle ScholarPubMed
Palermo, G, Joris, H, Devroey, P, Van Steirteghem, AC. Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte. Lancet 1992;340:1718.CrossRefGoogle ScholarPubMed
Palermo, G, Rosenwaks, Z. Assisted fertilization for male-factor infertility. Seminars in Reproductive Medicine 1995;13(01):3952.CrossRefGoogle Scholar
Hiramoto, Y. An analysis of the mechanism of fertilization by means of enucleation of sea urchin eggs. Experimental Cell Research 1962;28:323334.CrossRefGoogle ScholarPubMed
Hiramoto, Y. Microinjection of the live spermatozoa into sea urchin eggs. Experimental Cell Research 1962;27:416426.CrossRefGoogle ScholarPubMed
Uehara, T, Yanagimachi, R. Activation of hamster eggs by pricking. Journal of Experimental Zoology 1977;199:269274.CrossRefGoogle ScholarPubMed
Uehara, T, Yanagimachi, R. Behavior of nuclei of testicular, caput and cauda epididymal spermatozoa injected into hamster eggs. Biology of Reproduction 1977;16:315321.CrossRefGoogle ScholarPubMed
Goto, K, Kinoshita, A, Takuma, Y, Ogawa, K. Fertilisation of bovine oocytes by the injection of immobilised, killed spermatozoa. The Veterinary Record 1990;127:517520.Google ScholarPubMed
Iritani, A, Utsumi, K, Miyake, M, Hosoi, Y, Saeki, K. In vitro fertilization by a routine method and by micromanipulation. Annals of the New York Academy of Sciences 1988;541:583590.CrossRefGoogle ScholarPubMed
Lanzendorf, SE, Maloney, MK, Veeck, LL, Slusser, J, Hodgen, GD, Rosenwaks, Z. A preclinical evaluation of pronuclear formation by microinjection of human spermatozoa into human oocytes. Fertility and Sterility 1988;49:835842.CrossRefGoogle ScholarPubMed
Palermo, G, Joris, H, Devroey, P, Van Steirteghem, AC. Induction of acrosome reaction in human spermatozoa used for subzonal insemination. Human Reproduction 1992;7:248254.CrossRefGoogle ScholarPubMed
Lin, TP. Microinjection of mouse eggs. Science 1966;151:333337.CrossRefGoogle ScholarPubMed
Pereira, N, Cozzubbo, T, Cheung, S, Palermo, GD. Lessons learned in andrology: from intracytoplasmic sperm injection and beyond. Andrology 2016;4:757760.CrossRefGoogle ScholarPubMed
Palermo, G, Joris, H, Derde, MP, Camus, M, Devroey, P, Van Steirteghem, A. Sperm characteristics and outcome of human assisted fertilization by subzonal insemination and intracytoplasmic sperm injection. Fertility and Sterility 1993;59:826835.CrossRefGoogle ScholarPubMed
Palermo, GD, Neri, QV, Schlegel, PN, Rosenwaks, Z. Intracytoplasmic sperm injection (ICSI) in extreme cases of male infertility. PLoS One 2014;9:e113671.CrossRefGoogle ScholarPubMed
Palermo, GD, Neri, QV, Rosenwaks, Z. To ICSI or not to ICSI. Seminars in Reproductive Medicine 2015;33:92102.Google ScholarPubMed
Palermo, GD, Neri, QV, Takeuchi, T, Rosenwaks, Z. ICSI: where we have been and where we are going. Seminars in Reproductive Medicine 2009;27:191201.CrossRefGoogle ScholarPubMed
Palermo, G, Munne, S, Cohen, J. The human zygote inherits its mitotic potential from the male gamete. Human Reproduction 1994;9:12201225.CrossRefGoogle ScholarPubMed
Palermo, GD, Colombero, LT, Schattman, GL, Davis, OK, Rosenwaks, Z. Evolution of pregnancies and initial follow-up of newborns delivered after intracytoplasmic sperm injection. JAMA 1996;276:18931897.CrossRefGoogle ScholarPubMed
Practice Committees of the American Society for Reproductive Medicine, Society for Assisted Reproductive Technology. Intracytoplasmic sperm injection (ICSI) for non-male factor infertility: a committee opinion. Fertility and Sterility 2012;98:13951399.CrossRefGoogle Scholar
Palermo, GD, Cohen, J, Alikani, M, Adler, A, Rosenwaks, Z. Intracytoplasmic sperm injection: a novel treatment for all forms of male factor infertility. Fertility and Sterility 1995;63:12311240.CrossRefGoogle ScholarPubMed
Schlegel, PN, Cohen, J, Goldstein, M, Alikani, M, Adler, A, Gilbert, BR, Palermo, GD, Rosenwaks, Z. Cystic fibrosis gene mutations do not affect sperm function during in vitro fertilization with micromanipulation for men with bilateral congenital absence of vas deferens. Fertility and Sterility 1995;64:421426.CrossRefGoogle Scholar
Schlegel, PN, Palermo, GD, Alikani, M, Adler, A, Reing, AM, Cohen, J, Rosenwaks, Z. Micropuncture retrieval of epididymal sperm with in vitro fertilization: importance of in vitro micromanipulation techniques. Urology 1995;46:238241.CrossRefGoogle ScholarPubMed
Chan, PT, Palermo, GD, Veeck, LL, Rosenwaks, Z, Schlegel, PN. Testicular sperm extraction combined with intracytoplasmic sperm injection in the treatment of men with persistent azoospermia postchemotherapy. Cancer 2001;92:16321637.3.0.CO;2-I>CrossRefGoogle ScholarPubMed
Palermo, GD, Schlegel, PN, Sills, ES, Veeck, LL, Zaninovic, N, Menendez, S, Rosenwaks, Z. Births after intracytoplasmic injection of sperm obtained by testicular extraction from men with nonmosaic Klinefelter’s syndrome. The New England Journal of Medicine 1998;338:588590.CrossRefGoogle ScholarPubMed
O’Neill, CL, Parrella, A, Keating, D, Cheung, S, Rosenwaks, Z, Palermo, GD. A treatment algorithm for couples with unexplained infertility based on sperm chromatin assessment. Journal of Assisted Reproduction and Genetics 2018;35:19111917.CrossRefGoogle ScholarPubMed
Xie, P, Keating, D, Parrella, A, Cheung, S, Rosenwaks, Z, Goldstein, M, Palermo, GD. Sperm genomic integrity by TUNEL varies throughout the male genital tract. Journal of Urology 2020;203:802808.CrossRefGoogle Scholar
Yanagida, K, Katayose, H, Yazawa, H, Kimura, Y, Sato, A, Yanagimachi, H, Yanagimachi, R. Successful fertilization and pregnancy following ICSI and electrical oocyte activation. Human Reproduction 1999;14:13071311.CrossRefGoogle ScholarPubMed
Hoshi, K, Yanagida, K, Sato, A. Pretreatment of hamster oocytes with Ca2+ ionophore to facilitate fertilization by ooplasmic micro-injection. Human Reproduction 1992;7:871875.CrossRefGoogle ScholarPubMed
Yanagida, K, Morozumi, K, Katayose, H, Hayashi, S, Sato, A. Successful pregnancy after ICSI with strontium oocyte activation in low rates of fertilization. Reproductive Biomedicine Online 2006;13:801806.CrossRefGoogle ScholarPubMed
Tavalaee, M, Nomikos, M, Lai, FA, Nasr-Esfahani, MH. Expression of sperm PLCzeta and clinical outcomes of ICSI-AOA in men affected by globozoospermia due to DPY19L2 deletion. Reproductive Biomedicine Online 2018;36:348355.CrossRefGoogle ScholarPubMed
Wolny, YM, Fissore, RA, Wu, H, Reis, MM, Colombero, LT, Ergun, B, Rosenwaks, Z, Palermo, GD. Human glucosamine-6-phosphate isomerase, a homologue of hamster oscillin, does not appear to be involved in Ca2+ release in mammalian oocytes. Molecular Reproduction and Development 1999;52:277287.3.0.CO;2-0>CrossRefGoogle Scholar
Heindryckx, B, Van der Elst, J, De Sutter, P, Dhont, M. Treatment option for sperm- or oocyte-related fertilization failure: assisted oocyte activation following diagnostic heterologous ICSI. Human Reproduction 2005;20:22372241.CrossRefGoogle ScholarPubMed
Bonte, D, Ferrer-Buitrago, M, Dhaenens, L, Popovic, M, Thys, V, De Croo, I, De Gheselle, S, Steyaert, N, Boel, A, Vanden Meerschaut, F et al. Assisted oocyte activation significantly increases fertilization and pregnancy outcome in patients with low and total failed fertilization after intracytoplasmic sperm injection: a 17-year retrospective study. Fertility and Sterility 2019;112:266274.CrossRefGoogle ScholarPubMed
Gilchrist, RB, Lane, M, Thompson, JG. Oocyte-secreted factors: regulators of cumulus cell function and oocyte quality. Human Reproduction Update 2008;14:159177.CrossRefGoogle ScholarPubMed
Alikani, M, Palermo, G, Adler, A, Bertoil, M, Blake, M, Cohen, J. Intracytoplasmic sperm injection in dysmorphic human oocytes. Zygote 1995;3:283288.CrossRefGoogle ScholarPubMed
Serhal, PF, Ranieri, DM, Kinis, A, Marchant, S, Davies, M, Khadum, IM. Oocyte morphology predicts outcome of intracytoplasmic sperm injection. Human Reproduction 1997;12:12671270.CrossRefGoogle ScholarPubMed
Xia, P. Intracytoplasmic sperm injection: correlation of oocyte grade based on polar body, perivitelline space and cytoplasmic inclusions with fertilization rate and embryo quality. Human Reproduction 1997;12:17501755.CrossRefGoogle ScholarPubMed
Kahraman, S, Yakin, K, Donmez, E, Samli, H, Bahce, M, Cengiz, G, Sertyel, S, Samli, M, Imirzalioglu, N. Relationship between granular cytoplasm of oocytes and pregnancy outcome following intracytoplasmic sperm injection. Human Reproduction 2000;15:23902393.CrossRefGoogle ScholarPubMed
Meriano, JS, Alexis, J, Visram-Zaver, S, Cruz, M, Casper, RF. Tracking of oocyte dysmorphisms for ICSI patients may prove relevant to the outcome in subsequent patient cycles. Human Reproduction 2001;16:21182123.CrossRefGoogle ScholarPubMed
Yakin, K, Balaban, B, Isiklar, A, Urman, B. Oocyte dysmorphism is not associated with aneuploidy in the developing embryo. Fertility and Sterility 2007;88:811816.CrossRefGoogle Scholar
Swain, JE, Pool, TB. ART failure: oocyte contributions to unsuccessful fertilization. Human Reproduction Update 2008;14:431446.CrossRefGoogle ScholarPubMed
Eichenlaub-Ritter, U, Schmiady, H, Kentenich, H, Soewarto, D. Recurrent failure in polar body formation and premature chromosome condensation in oocytes from a human patient: indicators of asynchrony in nuclear and cytoplasmic maturation. Human Reproduction 1995;10:23432349.CrossRefGoogle ScholarPubMed
Parrella, A, Irani, M, Keating, D, Chow, S, Rosenwaks, Z, Palermo, GD. High proportion of immature oocytes in a cohort reduces fertilization, embryo development, pregnancy and live birth rates following ICSI. Reproductive Biomedicine Online 2019;39:580587.CrossRefGoogle Scholar
Kazem, R, Thompson, LA, Srikantharajah, A, Laing, MA, Hamilton, MP, Templeton, A. Cryopreservation of human oocytes and fertilization by two techniques: in-vitro fertilization and intracytoplasmic sperm injection. Human Reproduction 1995;10:26502654.CrossRefGoogle ScholarPubMed
Walls, ML, Ryan, JP, Keelan, JA, Hart, R. In vitro maturation is associated with increased early embryo arrest without impairing morphokinetic development of useable embryos progressing to blastocysts. Human Reproduction 2015;30:18421849.CrossRefGoogle ScholarPubMed
Hatirnaz, S, Ata, B, Hatirnaz, ES, Dahan, MH, Tannus, S, Tan, J, Tan, SL. Oocyte in vitro maturation: A systematic review. Turkish Journal of Obstetrics and Gynecology 2018;15:112125.CrossRefGoogle Scholar
Shu, YM, Zeng, HT, Ren, Z, Zhuang, GL, Liang, XY, Shen, HW, Yao, SZ, Ke, PQ, Wang, NN. Effects of cilostamide and forskolin on the meiotic resumption and embryonic development of immature human oocytes. Human Reproduction 2008;23:504513.CrossRefGoogle ScholarPubMed
Kirkegaard, K, Hindkjaer, JJ, Ingerslev, HJ. Human embryonic development after blastomere removal: a time-lapse analysis. Human Reproduction 2012;27:97105.CrossRefGoogle ScholarPubMed
Fragouli, E. Next generation sequencing for preimplantation genetic testing for aneuploidy: friend or foe? Fertility and Sterility 2018;109:606607.CrossRefGoogle ScholarPubMed
Coates, A, Kung, A, Mounts, E, Hesla, J, Bankowski, B, Barbieri, E, Ata, B, Cohen, J, Munne, S. Optimal euploid embryo transfer strategy, fresh versus frozen, after preimplantation genetic screening with next generation sequencing: a randomized controlled trial. Fertility and Sterility 2017;107:723730.e723.CrossRefGoogle ScholarPubMed
Forman, EJ, Hong, KH, Ferry, KM, Tao, X, Taylor, D, Levy, B, Treff, NR, Scott, RT, Jr. In vitro fertilization with single euploid blastocyst transfer: a randomized controlled trial. Fertility and Sterility 2013;100:100107.e101.CrossRefGoogle ScholarPubMed
Palmerola, K, Vitez, S, Amrane, S, Fischer, C, Forman, E. Minimizing mosaicism: assessing the impact of fertilization method on rate of mosaicism after next-generation sequencing (NGS) preimplantation genetic testing for aneuploidy (PGT-A). Journal of Assisted Reproduction and Genetics 2018;36.Google Scholar
Jun, SH, O’Leary, T, Jackson, KV, Racowsky, C. Benefit of intracytoplasmic sperm injection in patients with a high incidence of triploidy in a prior in vitro fertilization cycle. Fertility and Sterility 2006;86:825829.CrossRefGoogle Scholar
Zegers-Hochschild, F, Adamson, GD, de Mouzon, J, Ishihara, O, Mansour, R, Nygren, K, Sullivan, E, van der Poel, S, International Committee for Monitoring Assisted Reproductive Technology, World Health Organization. The International Committee for Monitoring Assisted Reproductive Technology (ICMART) and the World Health Organization (WHO) Revised Glossary on ART Terminology, 2009. Human reproduction 2009;24:26832687.CrossRefGoogle ScholarPubMed
Centers for Disease Control and Prevention. Key Statistics from the National Survey of Family Growth. 20 June 2017, Scholar
World Health Organization. WHO Laboratory Manual for the Examination and Processing of Human Semen. 5th ed., 2010. World Health Organization, Geneva.Google Scholar
Vassena, R, Eguizabal, C, Heindryckx, B, Sermon, K, Simon, C, van Pelt, AM, Veiga, A, Zambelli, F, ESHRE special interest group Stem Cells. Stem cells in reproductive medicine: ready for the patient? Human Reproduction 2015;30:2014–2021.Google Scholar
Hayashi, K, Ogushi, S, Kurimoto, K, Shimamoto, S, Ohta, H, Saitou, M. Offspring from oocytes derived from in vitro primordial germ cell-like cells in mice. Science 2012;338:971975.CrossRefGoogle ScholarPubMed
Hayashi, K, Ohta, H, Kurimoto, K, Aramaki, S, Saitou, M. Reconstitution of the mouse germ cell specification pathway in culture by pluripotent stem cells. Cell 2011;146:519532.CrossRefGoogle ScholarPubMed
Neuhaus, N, Schlatt, S. Stem cell-based options to preserve male fertility. Science 2019;363:12831284.CrossRefGoogle ScholarPubMed
Matoba, S, Zhang, Y. Somatic Cell Nuclear Transfer Reprogramming: Mechanisms and Applications. Cell Stem Cell 2018;23:471485.CrossRefGoogle ScholarPubMed
Pelosi, E, Forabosco, A, Schlessinger, D. Germ cell formation from embryonic stem cells and the use of somatic cell nuclei in oocytes. Annals of the New York Academy of Sciences 2011;1221:1826.CrossRefGoogle ScholarPubMed
Sato, T, Katagiri, K, Gohbara, A, Inoue, K, Ogonuki, N, Ogura, A, Kubota, Y, Ogawa, T. In vitro production of functional sperm in cultured neonatal mouse testes. Nature 2011;471:504507.CrossRefGoogle ScholarPubMed
Hayashi, K, Hikabe, O, Obata, Y, Hirao, Y. Reconstitution of mouse oogenesis in a dish from pluripotent stem cells. Nature Protocols 2017;12:17331744.CrossRefGoogle Scholar
Christin, JR, Beckert, MV. Origins and applications of CRISPR-mediated genome editing. The Einstein Journal of Biology and Medicine 2016;31:25.CrossRefGoogle Scholar
Makarova, KS, Grishin, NV, Shabalina, SA, Wolf, YI, Koonin, EV. A putative RNA-interference-based immune system in prokaryotes: computational analysis of the predicted enzymatic machinery, functional analogies with eukaryotic RNAi, and hypothetical mechanisms of action. Biology Direct 2006;1:7.CrossRefGoogle ScholarPubMed
Ma, H, Marti-Gutierrez, N, Park, S-W, Wu, J, Lee, Y, Suzuki, K, Koski, A, Ji, D, Hayama, T, Ahmed, R et al. Correction of a pathogenic gene mutation in human embryos. Nature 2017;548:413.CrossRefGoogle ScholarPubMed
Adashi, E, Cohen, IG. Heritable genome editing—edited eggs and sperm to the rescue? JAMA 2019;322:17541755.CrossRefGoogle ScholarPubMed
Wang, J, Parrella, A, Xie, P, Rosenwaks, Z, Palermo, GD. A step toward gene remodeling of mammalian spermatozoa by CRISPR-Cas9. Fertility and Sterility 2019;112:e262.CrossRefGoogle Scholar
Stein, R. Scientists attempt controversial experiment to edit DNA in human sperm using CRISPR. 2019. Scholar

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