title: Retinol improves in vitro differentiation of pre-pubertal mouse spermatogonial stem cells into sperm during the first wave of spermatogenesis.
abstract: Testicular tissue freezing has been proposed for fertility preservation in pre-pubertal boys. Thawed frozen testicular tissue must undergo a maturation process to restore sperm production. The purpose of the current study was to evaluate the ability of retinol to improve the in vitro differentiation of pre-pubertal mouse spermatogonial stem cells into sperm. Testes from pre-pubertal mice, aged 2.5 and 6.5 days post-partum, were cultured on agarose gel at a gas-liquid interphase for 34, 38 and 60 days (D) and for 16, 30 and 36 D respectively. Assessment of basal medium (BM) supplemented with retinol (RE) alone, FSH/LH alone or a combination of both, was performed. Stereological analyses and tissue lesion scoring were performed at the culture time points indicated above. Sperm production was quantified at D30 and D34 after mechanical dissection of the testicular tissues. FSH/LH significantly increased the percentage of round spermatids at D30 and D38, when compared to BM alone. However, RE significantly increased the percentages of round but also elongated spermatids at D30 and D34. Moreover, RE significantly increased the number of spermatozoa per milligram of tissue at D30 and D34 when compared to BM. Therefore, RE improved the in vitro production of spermatids and spermatozoa from pre-pubertal SSCs during the first wave of spermatogenesis. The use of RE could be a useful tool for in vitro spermatogenesis from pre-pubertal human 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: 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: Ludovic
authors.initials: L
authors.lastname: Dumont
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: Agathe
authors.initials: A
authors.lastname: Way
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: Biochemistry Laboratory, Rouen University Hospital, Institute for Biomedical Research, University of Rouen, Rouen, France.
authors.forename: Julien
authors.initials: J
authors.lastname: Wils
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: 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: D000375:Aging; D000818:Animals; D002454:Cell Differentiation; D015925:Cryopreservation; D005640:Follicle Stimulating Hormone / Q000378:metabolism; D007986:Luteinizing Hormone / Q000378:metabolism; D008297:Male; D051379:Mice; D013091:Spermatogenesis / Q000187:drug effects*; D013737:Testis / Q000166:cytology*; D014801:Vitamin A / Q000494:pharmacology*
chemical_list: D014801:Vitamin A; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone