Skip to main content Accessibility help

We use cookies to distinguish you from other users and to provide you with a better experience on our websites. Close this message to accept cookies or find out how to manage your cookie settings.

Close cookie message
×
Hostname: page-component-cd9895bd7-jkksz Total loading time: 0 Render date: 2024-12-28T13:40:47.892Z Has data issue: false hasContentIssue false

Chapter 30 - The Potential Future Applications of In Vitro Spermatogenesis in the Clinical Laboratory

from Section 4 - Laboratory Evaluation and Treatment of Male Infertility

Published online by Cambridge University Press:  06 December 2023

By
Swati Sharma  and
Stefan Schlatt
Edited by
Douglas T. Carrell ,
Alexander W. Pastuszak  and
James M. Hotaling
Douglas T. Carrell
Affiliation:
Utah Center for Reproductive Medicine
Alexander W. Pastuszak
Affiliation:
University of Utah
James M. Hotaling
Affiliation:
Utah Center for Reproductive Medicine
Get access

Summary

In this review we reflect on the many attempts highlighting key achievements in the field of in vitro spermatogenesis made so far. The research in this field is at a crucial juncture. The evolving technologies (like biofabricated 3D organoids, 3D bioprinting, microfluidics, or organ-on-a-chip) may offer excellent tools and in vitro testicular model systems to advance our understanding and bridge the existing knowledge gaps. Each of these culture systems offers unique advantages and may complement each other to address the common goal of achieving primate spermatogenesis in vitro. There are various possibilities and future scenarios for applying in vitro spermatogenesis as a tool for research and clinical applications in the future.

Keywords

in vitro spermatogenesismale infertilitygamete productionARTtesticular functionfertility restoration
Type
Chapter
Information
Men's Reproductive and Sexual Health Throughout the Lifespan
An Integrated Approach to Fertility, Sexual Function, and Vitality
 , pp. 229 - 243
Publisher: Cambridge University Press
Print publication year: 2023

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Anawalt, BD. Approach to male infertility and induction of spermatogenesis. J Clin Endocrinol Metab. 2013;98(9):3532–3542.Google Scholar
Levine, H, Jorgensen, N, Martino-Andrade, A, et al. Temporal trends in sperm count: a systematic review and meta regression analysis. Hum Reprod Update. 2017;23:646–659.Google Scholar
Sharpe, RM, Franks, S. Environment, lifestyle, and infertility: an inter-generational issue. Nat Cell Biol. 2002;s33–s40.CrossRefGoogle Scholar
Van Batavia, JP, Kolon, TF. Fertility in disorders of sex development. J Pediatr Urol. 2016;12(6):418–425.Google Scholar
Gomes, NL, Chetty, T, Jorgensen, A, Mitchell, RT. Disorders of sex development: novel regulators, impacts on fertility, and options for fertility preservation. Int J Mol Sci. 2020;21(7):2282.Google Scholar
Picton, HM, Wyns, C, Anderson, RA, et al. ESHRE task force on fertility preservation in severe diseases: a European perspective on testicular tissue cryopreservation for fertility preservation in prepubertal and adolescent boys. Hum Reprod. 2015;30(11):2463–2475.CrossRefGoogle Scholar
Weblink: www.urologyhealth.org/urologic-conditions/male-infertilityGoogle Scholar
Gassei, K, Orwig, KE. Experimental methods to preserve male fertility and treat male infertility. Fertil Steril. 2016;105(2):256–266.Google Scholar
Horwich, A, Shipley, J, Huddart, R. Testicular germ-cell cancer. Lancet. 2006;367:754–765.CrossRefGoogle ScholarPubMed
Rajpert-De Meyts, E, McGlynn, KA, Okamoto, K, Jewett, MAS, Bokemeyer, C. Testicular germ cell tumours. Lancet. 2016;387:1762–1774.Google Scholar
Sharma, S, Wistuba, J, Pock, T, Schlatt, S, Neuhaus, N. Spermatogonial stem cells: updates from specification to clinical relevance. Hum Reprod Update. 2019;25(3):275–297.Google Scholar
Goossens, E, Jahnukainen, K, Mitchell, RT, et al. Fertility preservation in boys: recent developments and new insights. Hum Reprod Open. 2020;2020(3):hoaa016.Google Scholar
Oncofertility Consortium. Homepage. https://oncofertility.northwestern.edu/Google Scholar
Schlatt, S, Ehmcke, J. Regulation of spermatogenesis: an evolutionary biologist’s perspective. Semin Cell Dev Biol. 2014;29:2–16.CrossRefGoogle ScholarPubMed
Tung, PS, Fritz, IB. Interactions of Sertoli cells with myoid cells in vitro. Biol Reprod. 1980;23:207–217.Google Scholar
Hadley, MA, Byers, SW, Suárez-Quian, CA, Kleinman, HK, Dym, M. Extracellular matrix regulates Sertoli cell differentiation, testicular cord formation, and germ cell development in vitro. J Cell Biol. 1985;101:1511–1522.Google Scholar
Kierszenbaum, AL, Crowell, JA, Shabanowitz, RB, DePhilip, RM, Tres, LL. Protein secretory patterns of rat Sertoli and peritubular cells are influenced by culture conditions. Biol Reprod. 1986;35:239–251.CrossRefGoogle ScholarPubMed
Tung, PS, Fritz, IB. Extracellular matrix components and testicular peritubular cells influence the rate and pattern of Sertoli cell migration in vitro. Dev Biol. 1986; 113:119–134.Google Scholar
Tung, PS, Fritz, IB. Morphogenetic restructuring and formation of basement membranes by Sertoli cells and testis peritubular cells in co-culture: inhibition of the morphogenetic cascade by cyclic AMP derivatives and by blocking direct cell contact. Dev Biol. 1987;120:139–153.CrossRefGoogle ScholarPubMed
Hofmann, MC, Narisawa, S, Hess, RA, Millan, JL. Immortalization of germ cells and somatic testicular cells using the SV40 large T antigen. Exp Cell Res. 1992;201:417–435.Google Scholar
Richardson, LL, Kleinman, HK, Dym, M. Basement membrane gene expression by Sertoli and peritubular myoid cells in vitro in the rat. Biol Reprod. 1995;52:320–330.Google Scholar
Schlatt, S, de Kretser, DM, Loveland, KL. Discriminative analysis of rat Sertoli and peritubular cells and their proliferation in vitro: evidence for follicle-stimulating hormonemediated contact inhibition of Sertoli cell mitosis. Biol Reprod. 1996;55:227–235.CrossRefGoogle ScholarPubMed
van der Wee, K, Hofmann, MC. An in vitro tubule assay identifies HGF as a morphogen for the formation of seminiferous tubules in the postnatal mouse testis. Exp Cell Res. 1999;252:175–185.Google Scholar
Hoeben, E, Swinnen, JV, Heyns, W, Verhoeven, G. Heregulins or neu differentiation factors and the interactions between peritubular myoid cells and Sertoli cells. Endocrinology. 1999;140:2216–2223.Google Scholar
El Ramy, R, Verot, A, Mazaud, S, Odet, F, Magre, S, Le Magueresse-Battistoni, B. Fibroblast growth factor (FGF) 2 and FGF9 mediate mesenchymal–epithelial interactions of peritubular and Sertoli cells in the rat testis. J Endocrinol. 2005;187:135–147.CrossRefGoogle ScholarPubMed
Gassei, K, Schlatt, S, Ehmcke, J. De novo morphogenesis of seminiferous tubules from dissociated immature rat testicular cells in xenografts. J Androl. 2006;27:611–618.Google Scholar
Marcon, L, Zhang, X, Hales, BF, Nagano, MC, Robaire, B. Development of a short term fluorescence-based assay to assess the toxicity of anticancer drugs on rat stem/progenitor spermatogonia in vitro. Biol Reprod. 2010;83:228–237.Google Scholar
Mincheva, M, Sandhowe-Klaverkamp, R, Wistuba, J, et al. Reassembly of adult human testicular cells: can testis cord-like structures be created in vitro? Mol Hum Reprod. 2018;24(2):55–63.CrossRefGoogle ScholarPubMed
von Kopylow, K, Schulze, W, Salzbrunn, A, et al. Mol Hum Reprod. 2018; 24(3):123–134.CrossRefGoogle Scholar
Mincheva, M, Wistuba, J, Brenker, C, Schlatt, S. Challenging human somatic testicular cell reassembly by protein kinase inhibtion: setting up a functional in vitro test system. Sci Rep. 2020;10(1):8935.Google Scholar
Iwanami, Y, Kobayashi, T, Kato, M, Hirabayashi, M, Hochi, S. Characteristics of rat round spermatids differentiated from spermatogonial cells during co-culture with Sertoli cells, assessed by flow cytometry, microinsemination and RT-PCR. Theriogenology. 2006;65:288–298.CrossRefGoogle ScholarPubMed
Xie, B, Qin, Z, Huang, B, et al. In vitro culture and differentiation of buffalo (Bubalus bubalis) spermatogonia. Reprod Domest Anim. 2010;45:275–282CrossRefGoogle ScholarPubMed
Tres, LL, Kierszenbaum, AL. Viability of rat spermatogenic cells in vitro is facilitated by their coculture with Sertoli cells in serum-free hormone-supplemented medium. Proc Natl Acad Sci USA. 1983;80:3377–3381.Google Scholar
Tesarik, J, Greco, E, Rienzi, L, et al. Differentiation of spermatogenic cells during in-vitro culture of testicular biopsy samples from patients with obstructive azoospermia: effect of recombinant follicle stimulating hormone. Hum Reprod. 1998a;13:2772–2781.Google Scholar
Tesarik, J, Guido, M, Mendoza, C, Greco, E. Human spermatogenesis in vitro: respective effects of follicle-stimulating hormone and testosterone on meiosis, spermiogenesis, and Sertoli cell apoptosis. J Clin Endocrinol Metab. 1998b;83:4467–4473.Google Scholar
Tesarik, J, Balaban, B, Isiklar, A, et al. In-vitro spermatogenesis resumption in men with maturation arrest: relationship with in-vivo blocking stage and serum FSH. Hum Reprod. 2000;15:1350–1354.Google Scholar
Sousa, M, Cremades, N, Alves, C, Silva, J, Barros, A. Developmental potential of human spermatogenic cells co-cultured with Sertoli cells. Hum Reprod. 2002;17:161–172.Google Scholar
Tanaka, A, Nagayoshi, M, Awata, S, Mawatari, Y, Tanaka, I, Kusunoki, H. Completion of meiosis in human primary spermatocytes through in vitro coculture with Vero cells. Fertil Steril. 2003;79:795–801.Google Scholar
Vigier, M, Weiss, M, Perrard, MH, Godet, M, Durand, P. The effects of FSH and of testosterone on the completion of meiosis and the very early steps of spermiogenesis of the rat: an in vitro study. J Mol Endocrinol. 2004;33:729–742.Google Scholar
Nagao, Y. Viability of meiotic prophase spermatocytes of rats is facilitated in primary culture of dispersed testicular cells on collagen gel by supplementing epinephrine or norepinephrine: evidence that meiotic prophase spermatocytes complete meiotic divisions in vitro. In Vitro Cell Dev Biol. 1989;25:1088–1098.Google Scholar
Magueresse-Battistoni, BL, Gérard, N, Jégou, B. Pachytene spermatocytes can achieve meiotic process in vitro. Biochem Biophys Res Commun. 1991;179(2):1115–1121.CrossRefGoogle ScholarPubMed
Cremades, N, Bernabeu, R, Barros, A, Sousa, M. In-vitro maturation of round spermatids using co-culture on Vero cells. Hum Reprod. 1999;14:1287–1293.CrossRefGoogle ScholarPubMed
Enders, GC, Henson, JH, Millette, CF. Sertoli cell binding to isolated testicular basement membrane. J Cell Biol. 1986;103:1109–1119.Google Scholar
Hadley, MA, Weeks, BS, Kleinman, HK, Dym, M. Laminin promotes formation of cord-like structures by Sertoli cells in vitro. Dev Biol. 1990;140:318–327.Google Scholar
Gassei, K, Ehmcke, J, Schlatt, S. Initiation of testicular tubulogenesis is controlled by neurotrophic tyrosine receptor kinases in a three-dimensional Sertoli cell aggregation assay. Reproduction. 2008;136:459–469.CrossRefGoogle Scholar
Yu, X, Hong, S, Moreira, EG, Faustman, EM. Improving in vitro Sertoli cell/gonocyte co-culture model for assessing male reproductive toxicity: lessons learned from comparisons of cytotoxicity versus genomic responses to phthalates. Toxicol Appl Pharmacol. 2009;239:325–336.CrossRefGoogle ScholarPubMed
Gassei, K, Ehmcke, J, Wood, MA, Walker, WH, Schlatt, S. Immature rat seminiferous tubules reconstructed in vitro express markers of Sertoli cell maturation after xenografting into nude mouse hosts. Mol Hum Reprod. 2010;16:97–110.Google Scholar
Pan, F, Chi, LF, Schlatt, S. Effects of nanostructures and mouse embryonic stem cells on in vitro morphogenesis of rat testicular cords. PLoS ONE. 2013;8:e60054.Google Scholar
Wegner, S, Hong, S, Yu, X, Faustman, EM. Preparation of rodent testis co-cultures. Curr Protoc Toxicol. 2013;Chapter 16:Unit 16.10.Google Scholar
Reuter, K, Ehmcke, J, Stukenborg, JB, et al. Reassembly of somatic cells and testicular organogenesis in vitro. Tissue Cell. 2014;46:86–96.Google Scholar
Potter, SJ, DeFalco, T. Using ex vivo upright droplet cultures of whole fetal organs to study developmental processes during mouse organogenesis. J Vis Exp. 2015;e53262.Google Scholar
Harris, S, Hermsen, SAB, Yu, XZ, Hong, SW, Faustman, EM. Comparison of toxicogenomic responses to phthalate ester exposure in an organotypic testis co-culture model and responses observed in vivo. Reprod Toxicol. 2015;58:149–159.CrossRefGoogle Scholar
Harris, S, Shubin, SP, Wegner, S, et al. The presence of macrophages and inflammatory responses in an in vitro testicular co-culture model of male reproductive development enhance relevance to in vivo conditions. Toxicol In Vitro. 2016;36:210–215.Google Scholar
Sakib, S, Uchida, A, Valenzuela-Leon, P, et al. Formation of organotypic testicular organoids in microwell culture. Biol Reprod. 2019;100(6):1648–1660.Google Scholar
Edmonds, ME, Woodruff, TK. Testicular organoid formation is a property of immature somatic cells, which self-assemble and exhibit long-term hormone-responsive endocrine function. Biofabrication. 2020;12(4):045002.Google Scholar
Baert, Y, Ruetschle, I, Cools, W, et al. A multi-organ chip co-culture of liver and testis equivalents: a first step toward a systemic male reprotoxicity model. Hum Reprod. 2020;35(5):1029–1044.Google Scholar
Zenzes, MT, Engel, W. The capacity of testicular cells of the postnatal rat to reorganize into histotypic structures. Differentiation. 1981;20:157–161.Google Scholar
Lee, DR, Kaproth, MT, Parks, JE. In vitro production of haploid germ cells from fresh or frozen-thawed testicular cells of neonatal bulls. Biol Reprod. 2001;65:873–878.Google Scholar
Lee, DR, Kim, K-S, Yang, YH, et al. Isolation of male germ stem cell-like cells from testicular tissue of non-obstructive azoospermic patients and differentiation into haploid male germ cells in vitro. Hum Reprod. 2006a;21:471–476.Google Scholar
Lee, JH, Kim, HJ, Kim, H, Lee, SJ, Gye, MC. In vitro spermatogenesis by threedimensional culture of rat testicular cells in collagen gel matrix. Biomaterials. 2006b;27:2845–2853.Google Scholar
Legendre, A, Froment, P, Desmots, S, Lecomte, A, Habert, R, Lemazurier, E. An engineered 3D blood-testis barrier model for the assessment of reproductive toxicity potential. Biomaterials. 2010;31:4492–4505.Google Scholar
Yokonishi, T, Sato, T, Katagiri, K, Komeya, M, Kubota, Y, Ogawa, T. In vitro reconstruction of mouse seminiferous tubules supporting germ cell differentiation. Biol Reprod. 2013;89:11–16.Google Scholar
Zhang, J, Hatakeyama, J, Eto, K, Abe, S. Reconstruction of a seminiferous tubule-like structure in a 3 dimensional culture system of re-aggregated mouse neonatal testicular cells within a collagen matrix. Gen Comp Endocrinol. 2014;205:121–132.Google Scholar
Reda, A, Hou, M, Landreh, L, et al. In vitro spermatogenesis: optimal culture conditions for testicular cell survival, germ cell differentiation, and steroidogenesis in rats. Front Endocrinol (Lausanne). 2014;5:21.Google Scholar
Huleihel, M, Nourashrafeddin, S, Plant, TM. Application of three-dimensional culture systems to study mammalian spermatogenesis, with an emphasis on the rhesus monkey (Macaca mulatta). Asian J Androl. 2015;17:972–980.Google Scholar
Baert, Y, De Kock, J, Alves-Lopes, JP, Söder, O, Stukenborg, J-B, Goossens, E. Primary human testicular cells self-organize into organoids with testicular properties. Stem Cell Reports. 2017;1:30–38.Google Scholar
Baert, Y, Goossens, E. Preparation of scaffolds from decellularized testicular matrix. Methods Mol Biol. 2018;1577:121–127.Google Scholar
Baert, Y, Rombaut, C, Goossens, E. Scaffold-based and scaffold-free testicular organoids from primary human testicular cells. Methods Mol Biol. 2019;1576:283–290.Google Scholar
Baert, Y, Stukenborg, JB, Landreh, M, et al. Derivation and characterization of a cytocompatible scaffold from human testis. Hum Reprod. 2015;30:256–267.Google Scholar
Alves-Lopes, JP, Soder, O, Stukenborg, JB. Testicular organoid generation by a novel in vitro three-layer gradient system. Biomaterials. 2017;130:76–89.Google Scholar
Lee, J-H, Gye, MC, Choi, KW, et al. In vitro differentiation of germ cells from nonobstructive azoospermic patients using three-dimensional culture in a collagen gel matrix. Fertil Steril. 2007;87:824–833.Google Scholar
Lee, JH, Oh, JH, Lee, JH, Kim, MR, Min, CK. Evaluation of in vitro spermatogenesis using poly(D,L-lactic-co-glycolic acid) (PLGA)-based macroporous biodegradable scaffolds. J Tissue Eng Regen Med. 2011;5:130–137.Google Scholar
Zhang, X, Wang, L, Zhang, X, et al. The use of KnockOut serum replacement (KSR) in three dimensional rat testicular cells coculture model: an improved male reproductive toxicity testing system. Food Chem Toxicol. 2017;106:487–495.Google Scholar
Stukenborg, JB, Wistuba, J, Luetjens, CM, et al. Coculture of spermatogonia with somatic cells in a novel three-dimensional soft-agar-culture-system. J Androl. 2008;29:312–329.Google Scholar
Stukenborg, J-B, Schlatt, S, Simoni, M, et al. New horizons for in vitro spermatogenesis? An update on novel three-dimensional culture systems as tools for meiotic and post-meiotic differentiation of testicular germ cells. Mol Hum Reprod. 2009;15:521–529.Google Scholar
Pendergraft, SS, Sadri-Ardekani, H, Atala, A, Bishop, CE. Three-dimensional testicular organoid: a novel tool for the study of human spermatogenesis and gonadotoxicity in vitro dagger. Biol Reprod. 2017;96:720–732.Google Scholar
Elhija, MA, Lunenfeld, E, Schlatt, S, Huleihel, M. Differentiation of murine male germ cells to spermatozoa in a soft agar culture system. Asian J Androl. 2012;14:285–293.Google Scholar
Alves-Lopes, JP, Stukenborg, JB. Testicular organoids: a new model to study the testicular microenvironment in vitro? Hum Reprod Update. 2018;24(2):176–191.Google Scholar
Sharma, S, Venzac, B, Burgers, T, Le Gac, S, Schlatt, S. Microfluidics in male reproduction: is ex vivo culture of primate testis tissue a future strategy for ART or toxicology research? Mol Hum Reprod. 2020;26(3):179–192.Google Scholar
Sakib, S, Goldsmith, T, Voigt, A, Dobrinski, I. Testicular organoids to study cell-cell interactions in the mammalian testis. Andrology. 2019; 8(4):835–841.Google Scholar
Komeya, M, Sato, T, Ogawa, T. In vitro spermatogenesis: a century-long research journey, still half way around. Reprod Med Biol. 2018;17(4):407–420.Google Scholar
Gargus, ES, Rogers, HB, McKinnon, KE, Edmonds, ME, Woodruff, TK. Engineered reproductive tissues. Nat Biomed Eng. 2020;4(4):381–393.Google Scholar
Szczepny, A, Hogarth, CA, Young, J, Loveland, KL. Identification of Hedgehog signaling outcomes in mouse testis development using a hanging drop‐culture system. Biol Reprod. 1999;80:258–263.Google Scholar
Jørgensen, A, Young, J, Nielsen, JE, et al. Hanging drop cultures of human testis and testis cancer samples: a model used to investigate activin treatment effects in a preserved niche. Br J Cancer. 2014;110:2604–2614.Google Scholar
Jørgensen, A, Nielsen, JE, Perlman, S, et al. Ex vivo culture of human fetal gonads: manipulation of meiosis signalling by retinoic acid treatment disrupts testis development. Hum Reprod. 2015;30:2351–2363.Google Scholar
Champy, CH. De la méthode de culture des tissus. VI. Le testicule. Arch Zool Exptl Gen. 1920;60:461–500.Google Scholar
Martinovitch, PN. The development in vitro of the mammalian gonad. Ovary and ovogenesis. Proc R Soc B Biol Sci. 1938;125:232–249.Google Scholar
Trowell, OA. The culture of mature organs in a synthetic medium. Exp Cell Res. 1959;16:118–147.CrossRefGoogle Scholar
Steinberger, E, Steinberger, A, Perloff, WH. Studies on growth in organ culture of testicular tissue from rats of various ages. Anat Rec. 1964a;148:581–589.Google Scholar
Steinberger, E, Steinberger, A, Perloff, WH. Initiation of spermatogenesis in vitro. Endocrinology. 1964b;74:788–792.Google Scholar
Steinberger, A, Steinberger, E, Perloff, WH. Mammalian testis in organ culture. Exp Cell Res. 1964c;36:19–27.Google Scholar
Steinberger, A, Steinberger, E. Differentiation of rat seminiferous epithelium in organ culture. J Reprod Fertil. 1965;9:243–248.Google Scholar
Steinberger, A, Steinberger, E. Factors affecting spermatogenesis in organ cultures of mammalian testes. J Reprod Fertil. 1967;Suppl. 2:117–124.Google Scholar
Matte, R, Sasaki, M. Autoradiographic evidence of human male germ‐cell differentiation in vitro. Cytologia. 1971;36:298–303.Google Scholar
Boitani, C, Politi, MG, Menna, T. Spermatogonial cell proliferation in organ culture of immature rat testis. Biol Reprod. 1993;48:761–767.Google Scholar
Schlatt, S, Zhengwei, Y, Meehan, T, de Kretser, DM, Loveland, KL. Application of morphometric techniques to postnatal rat testes in organ culture: insights into testis growth. Cell Tissue Res. 1999;298:335–343.Google Scholar
Meehan, T, Schlatt, S, O’Bryan, M, de Kretser, DM, Loveland, KL. Regulation of germ cell and Sertoli cells development by activin, follistatin and FSH. Dev Biol. 2000;220:225–237.Google Scholar
Suzuki, S, Sato, K. The fertilising ability of spermatogenic cells derived from cultured mouse immature testicular tissue. Zygote. 2003;11:307–316.Google Scholar
Lambrot, R, Coffigny, H, Pairault, C, et al. Use of organ culture to study the human fetal testis development: effect of retinoic acid. J Clin Endocrinol Metab. 2006;91:2696–2703.Google Scholar
Roulet, V, Denis, H, Staub, C, et al. Human testis in organotypic culture: application for basic or clinical research. Hum Reprod. 2006;21:1564–1575.Google Scholar
Gohbara, A, Katagiri, K, Sato, T, et al. In vitro murine spermatogenesis in an organ culture system. Biol Reprod. 2010;83:261–267.Google Scholar
Sato, T, Katagiri, K, Gohbara, A, et al. In vitro production of functional sperm in cultured neonatal mouse testes. Nature. 2011a;471:504–507.Google Scholar
Sato, T, Katagiri, K, Yokonishi, T, et al. In vitro production of fertile sperm from murine spermatogonial stem cell lines. Nat Commun. 2011b;2:472.Google Scholar
Dumont, L, Oblette, A, Rondanino, C, et al. Vitamin A prevents round spermatid nuclear damage and promotes the production of motile sperm during in vitro maturation of vitrified prepubertal mouse testicular tissue. Mol Hum Reprod. 2016;22:819–832.Google Scholar
Liu, F, Cai, C, Wu, X, et al. Effect of KnockOut serum replacement on germ cell development of immature testis tissue culture. Theriogenology. 2016;85:193–199.Google Scholar
Perrard, MH, Sereni, N, Schluth-Bolard, C, et al. Complete human and rat ex vivo spermatogenesis from fresh or frozen testicular tissue. Biol Reprod. 2016;95:89.Google Scholar
Komeya, M, Kimura, H, Nakamura, H, et al. Long-term ex vivo maintenance of testis tissues producing fertile sperm in a microfluidic device. Sci Rep. 2016;6:21472.Google Scholar
Komeya, M, Hayashi, K, Nakamura, H, et al. Pumpless microfluidic system driven by hydrostatic pressure induces and maintains mouse spermatogenesis in vitro. Sci Rep. 2017;7(1):15459.Google Scholar
Reda, A, Albalushi, H, Montalvo, SC, et al. Knock-out serum replacement and melatonin effects on germ cell differentiation in murine testicular explant cultures. Ann Biomed Eng. 2017;45:1783–1794.Google Scholar
Rondanino, C, Maouche, A, Dumont, L, Oblette, A, Rives, N. Establishment, maintenance and functional integrity of the blood-testis barrier in organotypic cultures of fresh and frozen/thawed prepubertal mouse testes. Mol Hum Reprod. 2017;23:304–320.Google Scholar
Stukenborg, J-B, Alves-Lopes, JP, Kurek, M, et al. Spermatogonial quantity in human prepubertal testicular tissue collected for fertility preservation prior to potentially sterilizing therapy. Hum Reprod. 2018;33(9):1677–1683.Google Scholar
Medrano, JV, Vilanova-Pérez, T, Fornés-Ferrer, V, et al. Influence of temperature, serum, and gonadotropin supplementation in short- and long-term organotypic culture of human immature testicular tissue. Fertil Steril. 2018;110(6):1045–1057.Google Scholar
Yamanaka, H, Komeya, M, Nakamura, H, et al. A monolayer microfluidic device supporting mouse spermatogenesis with improved visibility. Biochem BioPhys Res Commun. 2018;500(4):885–891.Google Scholar
Portela, JMD, de Winter-Korver, CM, van Daalen, SKM, et al. Assessment of fresh and cryopreserved testicular tissues from (pre)pubertal boys during organ culture as a strategy for in vitro spermatogenesis. Hum Reprod. 2019;34(12):2443–2455.Google Scholar
Ghatnekar, R, Lima‐De‐faria, A, Rubin, S, Menander, K. Development of human male meiosis in vitro. Hereditas. 1974;78:265–271.Google Scholar
Eddy, EM, Kahri, AI. Cell associations and surface features in cultures of juvenile rat seminiferous tubules. Anat Rec. 1976;185:333–357.Google Scholar
Aizawa, S, Nishimune, Y. In‐vitro differentiation of type A spermatogonia in mouse cryptorchid testis. J Reprod Fertil. 1979;56:99–104.Google Scholar
Curtis, D. In vitro differentiation of diakinesis figures in human testis. Hum Genet. 1981;59:406–411.Google Scholar
Nishimune, YM, Osaka, M. In vitro differentiation mouse cryptorchid of type a spermatogonia from testes in serum‐free. Biol Reprod. 1983;28:1217–1223.Google Scholar
Parvinen, M, Wright, WW, Phillips, DM, Mather, NA, Musto, NA, Bardin, CW. Spermatogenesis in vitro: completion of meiosis and early spermiogenesis. Endocrinology. 1983;112:1150–1152.Google Scholar
Toppari, J, Mali, P, Eerola, E. Rat spermatogenesis in vitro traced by quantitative flow cytometry. J Histochem Cytochem. 1986;34:1029–1035.Google Scholar
Kim, KJ, Kim, BG, Kim, YH, et al. In vitro spermatogenesis using bovine testis tissue culture techniques. Tissue Eng Regener Med. 2015;12:314–323.Google Scholar
Arkoun, B, Dumont, L, Milazzo, JP, et al. Retinol improves in vitro differentiation of pre-pubertal mouse spermatogonial stem cells into sperm during the first wave of spermatogenesis. PLoS ONE. 2015;10:e0116660.Google Scholar
Dumont, L, Arkoun, B, Jumeau, F, Milazzo, J-F, Bironneau, A, Liot, D, Wils, J, Rondanino, C, Rives, N. Assessment of the optimal vitrification protocol for pre-pubertal mice testes leading to successful in vitro production of flagellated spermatozoa. Andrology. 2015; 3(3):611–625.Google Scholar
Sato, T, Katagiri, K, Kojima, K, Komeya, M, Yao, M, Ogawa, T. In vitro spermatogenesis in explanted adult mouse testis tissues. PLoS ONE. 2015;10:e0130171.Google Scholar
de Michele, F, Poels, J, Weerens, L, et al. Preserved seminiferous tubule integrity with spermatogonial survival and induction of Sertoli and Leydig cell maturation after long-term organotypic culture of prepubertal human testicular tissue. Hum Reprod. 2017a;32:32–45.Google Scholar
de Michele, F, Vermeulen, M, Wyns, C. Fertility restoration with spermatogonial stem cells. Curr Opin Endocrinol Diabetes Obes. 2017b;24:424–431.Google Scholar
Gholami, K, Vermeulen, M, Del Vento, F, de Michele, F, Giudice, MG, Wyns, C. The air-liquid interface culture of the mechanically isolated seminiferous tubules embedded in agarose or alginate improves in vitro spermatogenesis at the expense of attenuating their integrity. In Vitro Cell Dev Biol Anim. 2020;56(3):261–270.Google Scholar
Yokonishi, T, Sato, T, Komeya, M, et al. Offspring production with sperm grown in vitro from cryopreserved testis tissues. Nat Commun. 2014;5:4320.Google Scholar
Steinberger, A, Steinberger, E. In vitro culture of rat testicular cells. Exptl Cell Res. 1966;44:443–452.Google Scholar
Rassoulzadegan, M, Paquis-Flucklinger, V, Bertino, B, et al. Transmeiotic differentiation of male germ cells in culture. Cell. 1993; 75(5):997–1006.Google Scholar
Marh, J, Tres, LL, Yamazaki, Y, Yanagimachi, R, Kierszenbaum, AL. Mouse round spermatids developed in vitro from preexisting spermatocytes can produce normal offspring by nuclear injection into in vivo‐developed mature oocytes. Biol Reprod. 2003;69:169–176.Google Scholar
Hofmann, MC, Hess, RA, Goldberg, E, Millán, JL. Immortalized germ cells undergo meiosis in vitro. PNAS, 1994; 91(12): 5533–5537.Google Scholar
Hasegawa, H, Terada, Y, Ugajin, T, Yaegashi, N, Sato, K. A novel culture system for mouse spermatid maturation which produces elongating spermatids capable of inducing calcium oscillation during fertilization and embryonic development. J Assist Reprod Genet. 2010;27(9–10):565–570.Google Scholar
Izadyar, F, Den Ouden, K, Creemers, LB, Posthuma, G, Parvinen, M, De Rooij, DG. Proliferation and differentiation of bovine type A spermatogonia during long-term culture. Biol Reprod. 2003;68(1):272–281.Google Scholar
Grinspon, RP, Rey, RA. Molecular characterization of XX maleness. Int J Mol Sci. 2019;20:6089.Google Scholar
Steinberger, A, Steinberger, E, Perloff, WH. Growth of rat testes fragments in organ culture. Fed Proc. 1963;22:372.Google Scholar
Steinberger, A, Steinberger, E. Stimulatory effect of vitamins and glutamine on the differentiation of germ cells in rat testes organ culture grown in chemically defined media. Exp Cell Res. 1966;44:429–435.Google Scholar
Parvinen, M, Vanha-Perttula, T. Identification and enzyme quantification of the stages of the seminiferous epithelial wave in the rat. Anat Rec. 1972;174:435–449.Google Scholar
Alves-Lopes, JP, Söder, O, Stukenborg, JB. Use of a three-layer gradient system of cells for rat testicular organoid generation. Nat Protoc. 2018;13:248–259.Google Scholar

Save book to Kindle

To save this book to your Kindle, first ensure [email protected] is added to your Approved Personal Document E-mail List under your Personal Document Settings on the Manage Your Content and Devices page of your Amazon account. Then enter the ‘name’ part of your Kindle email address below. Find out more about saving to your Kindle.

Note you can select to save to either the @free.kindle.com or @kindle.com variations. ‘@free.kindle.com’ emails are free but can only be saved to your device when it is connected to wi-fi. ‘@kindle.com’ emails can be delivered even when you are not connected to wi-fi, but note that service fees apply.

Find out more about the Kindle Personal Document Service.

Available formats
×

Save book to Dropbox

To save content items to your account, please confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your account. Find out more about saving content to Dropbox.

Available formats
×

Save book to Google Drive

To save content items to your account, please confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your account. Find out more about saving content to Google Drive.

Available formats
×