title: Effects of vitamin A on in vitro maturation of pre-pubertal mouse spermatogonial stem cells.
abstract: Testicular tissue cryopreservation is the only potential option for fertility preservation in pre-pubertal boys exposed to gonadotoxic treatment. Completion of spermatogenesis after in vitro maturation is one of the future uses of harvested testicular tissue. The purpose of the current study was to evaluate the effects of vitamin A on in vitro maturation of fresh and frozen-thawed mouse pre-pubertal spermatogonial stem cells in an organ culture system. Pre-pubertal CD1 mouse fresh testes were cultured for 7 (D7), 9 (D9) and 11 (D11) days using an organ culture system. Basal medium was supplemented with different concentrations of retinol (Re) or retinoic acid (RA) alone or in combination. Seminiferous tubule morphology (tubule diameter, intra-tubular cell type), intra-tubular cell death and proliferation (PCNA antibody) and testosterone level were assessed at D7, D9 and D11. Pre-pubertal mouse testicular tissue were frozen after a soaking temperature performed at -7 °C, -8 °C or -9 °C and after thawing, were cultured for 9 days, using the culture medium preserving the best fresh tissue functionality. Retinoic acid at 10(-6)M and retinol at 3.3.10(-7)M, as well as retinol 10(-6)M are favourable for seminiferous tubule growth, maintenance of intra-tubular cell proliferation and germ cell differentiation of fresh pre-pubertal mouse spermatogonia. Structural and functional integrity of frozen-thawed testicular tissue appeared to be well-preserved after soaking temperature at -8 °C, after 9 days of organotypic culture using 10(-6)M retinol. RA and Re can control in vitro germ cell proliferation and differentiation. Re at a concentration of 10(-6)M maintains intra-tubular cell proliferation and the ability of spermatogonia to initiate spermatogenesis in fresh and frozen pre-pubertal mouse testicular tissue using a soaking temperature at -8 °C. Our data suggested a possible human application for in vitro maturation of cryopreserved pre-pubertal testicular tissue.
authors.affiliation: EA 4308 "Gametogenesis and Gamete Quality", Reproductive Biology Laboratory - CECOS, Rouen University Hospital, Institute for Biomedical Research, University of Rouen, Rouen, France.
authors.forename: Albanne
authors.initials: A
authors.lastname: Travers
authors.affiliation: EA 4308 "Gametogenesis and Gamete Quality", Reproductive Biology Laboratory - CECOS, Rouen University Hospital, Institute for Biomedical Research, University of Rouen, Rouen, France.
authors.forename: Brahim
authors.initials: B
authors.lastname: Arkoun
authors.affiliation: EA 4308 "Gametogenesis and Gamete Quality", Reproductive Biology Laboratory - CECOS, Rouen University Hospital, Institute for Biomedical Research, University of Rouen, Rouen, France.
authors.forename: Athmane
authors.initials: A
authors.lastname: Safsaf
authors.affiliation: EA 4308 "Gametogenesis and Gamete Quality", Reproductive Biology Laboratory - CECOS, Rouen University Hospital, Institute for Biomedical Research, University of Rouen, Rouen, France.
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Milazzo
authors.affiliation: EA 4308 "Gametogenesis and Gamete Quality", Reproductive Biology Laboratory - CECOS, Rouen University Hospital, Institute for Biomedical Research, University of Rouen, Rouen, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Absyte
authors.affiliation: EA 4308 "Gametogenesis and Gamete Quality", Reproductive Biology Laboratory - CECOS, Rouen University Hospital, Institute for Biomedical Research, University of Rouen, Rouen, France.
authors.forename: Amandine
authors.initials: A
authors.lastname: Bironneau
authors.affiliation: EA 4308 "Gametogenesis and Gamete Quality", Reproductive Biology Laboratory - CECOS, Rouen University Hospital, Institute for Biomedical Research, University of Rouen, Rouen, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Perdrix
authors.affiliation: EA 4308 "Gametogenesis and Gamete Quality", Department of Urology, Rouen University Hospital, Institute for Biomedical Research, University of Rouen, Rouen, France.
authors.forename: Louis
authors.initials: L
authors.lastname: Sibert
authors.affiliation: EA 4308 "Gametogenesis and Gamete Quality", Reproductive Biology Laboratory - CECOS, Rouen University Hospital, Institute for Biomedical Research, University of Rouen, Rouen, France.
authors.forename: Bertrand
authors.initials: B
authors.lastname: Macé
authors.affiliation: Biochemistry Laboratory, Rouen University Hospital, Institute for Biomedical Research, University of Rouen, Rouen, France.
authors.forename: Bruno
authors.initials: B
authors.lastname: Cauliez
authors.affiliation: EA 4308 "Gametogenesis and Gamete Quality", Reproductive Biology Laboratory - CECOS, Rouen University Hospital, Institute for Biomedical Research, University of Rouen, Rouen, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Rives
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D016923:Cell Death / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D008297:Male; D008540:Meiosis / Q000187:drug effects / Q000502:physiology; D051379:Mice; D012741:Sexual Maturation / Q000187:drug effects*; D013091:Spermatogenesis / Q000187:drug effects / Q000502:physiology; D013093:Spermatogonia / Q000166:cytology* / Q000187:drug effects*; D013737:Testis / Q000187:drug effects / Q000378:metabolism; D013997:Time Factors; D014801:Vitamin A / Q000494:pharmacology*
chemical_list: D014801:Vitamin A