title: In vitro production of functional sperm in cultured neonatal mouse testes.
abstract: Spermatogenesis is one of the most complex and longest processes of sequential cell proliferation and differentiation in the body, taking more than a month from spermatogonial stem cells, through meiosis, to sperm formation. The whole process, therefore, has never been reproduced in vitro in mammals, nor in any other species with a very few exceptions in some particular types of fish. Here we show that neonatal mouse testes which contain only gonocytes or primitive spermatogonia as germ cells can produce spermatids and sperm in vitro with serum-free culture media. Spermatogenesis was maintained over 2 months in tissue fragments positioned at the gas-liquid interphase. The obtained spermatids and sperm resulted in healthy and reproductively competent offspring through microinsemination. In addition, neonatal testis tissues were cryopreserved and, after thawing, showed complete spermatogenesis in vitro. Our organ culture method could be applicable through further refinements to a variety of mammalian species, which will serve as a platform for future clinical application as well as mechanistic understanding of spermatogenesis.
authors: Department of Urology, Yokohama City University Graduate School of Medicine, Yokohama 236-0004, Japan.
authors: Takuya
authors: T
authors: Sato
authors: Kumiko
authors: K
authors: Katagiri
authors: Ayako
authors: A
authors: Gohbara
authors: Kimiko
authors: K
authors: Inoue
authors: Narumi
authors: N
authors: Ogonuki
authors: Atsuo
authors: A
authors: Ogura
authors: Yoshinobu
authors: Y
authors: Kubota
authors: Takehiko
authors: T
authors: Ogawa
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D015925:Cryopreservation / Q000379:methods; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D005260:Female; D005298:Fertility / Q000502:physiology; D005307:Fertilization in Vitro; D007248:Infertility, Male / Q000517:prevention & control; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009924:Organ Culture Techniques / Q000379:methods*; D012098:Reproduction / Q000502:physiology; D013087:Spermatids / Q000187:drug effects / Q000254:growth & development / Q000502:physiology; D013091:Spermatogenesis / Q000187:drug effects; D013093:Spermatogonia / Q000187:drug effects / Q000254:growth & development / Q000502:physiology; D013094:Spermatozoa / Q000187:drug effects / Q000254:growth & development / Q000502:physiology*; D013737:Testis / Q000187:drug effects / Q000254:growth & development* / Q000502:physiology*
chemical_list: D016895:Culture Media, Serum-Free