NDLI: Does soaking temperature during controlled slow freezing of pre-pubertal mouse testes influence course of in vitro spermatogenesis?

Content Provider	Springer Nature Link
Author	Arkoun, Brahim Dumont, Ludovic Milazzo, Jean Pierre Rondani, Christine Bironneau, Amandine Wils, Julien Rives, Nathalie
Copyright Year	2015
Abstract	The banking of testicular tissue before highly gonadotoxic treatment is a prerequisite for the preservation of fertility in pre-pubertal boys not yet producing sperm. The aim of the current study is to evaluate the impact of a soaking temperature performed at −7 °C, −8 °C or −9 °C on the ability of frozen-thawed mouse spermatogonial stem cells (SSCs) to generate haploid germ cells after in vitro maturation. Testes of 6.5-day-old post-partum CD-1 mice were cryopreserved by using a controlled slow freezing protocol with soaking at −7 °C, −8 °C or −9 °C. Frozen-thawed pre-pubertal testicular tissues were cultured in vitro on agarose gel for 30 days. Histological evaluations were performed and flagellated late spermatids were counted after mechanical dissection of the cultured tissues. The differentiation of frozen SSCs into elongated spermatids was more efficient after treatment at −9 °C than at −7 °C and −8 °C. After dissection, flagellated late spermatids were observed by using Shorr staining. The number of flagellated late spermatids was significantly decreased after slow freezing when compared with a fresh tissue control. Therefore, the soaking temperature during slow freezing of pre-pubertal mouse testicular tissue might positively influence the course of in vitro spermatogenesis. Our slow freezing protocol with a soaking temperature at −9 °C was the optimal condition in terms of the achievement of in vitro spermatogenesis with a higher production of elongated spermatids, although the effectiveness of the maturation process was reduced compared with the fresh tissue control.
Starting Page	661
Ending Page	674
Page Count	14
File Format	PDF
ISSN	0302766X
Journal	Cell and Tissue Research
Volume Number	364
Issue Number	3
e-ISSN	14320878
Language	English
Publisher	Springer Berlin Heidelberg
Publisher Date	2015-12-29
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Controlled slow freezing In vitro spermatogenesis Soaking temperature Spermatogonial stem cells Testicular tissue Human Genetics Proteomics Molecular Medicine
Content Type	Text
Resource Type	Article
Subject	Cell Biology Histology Pathology and Forensic Medicine

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