Improving Freezing Protocols and Organotypic Culture: A Histological Study on Rat Prepubertal Testicular Tissue

Testicular tissue freezing before gonadotoxic treatments allows the preservation of fertility for children suffering from cancer. Recently, the testis organ culture method was presented as a relevant method to restore the fertility of these patients. However, the yield of spermatozoa production is l...

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Bibliographic Details
Published in:Annals of biomedical engineering 2021-01, Vol.49 (1), p.203-218
Main Authors: Saulnier, Justine, Oblette, Antoine, Delessard, Marion, Dumont, Ludovic, Rives, Aurélie, Rives, Nathalie, Rondanino, Christine
Format: Article
Language:English
Subjects:
Animals
Biochemistry
Biological and Medical Physics
Biomedical and Life Sciences
Biomedical Engineering and Bioengineering
Biomedicine
Biophysics
Cell culture
Cell death
Cell differentiation
Cell Proliferation
Classical Mechanics
Cryopreservation
Fertility
Freezing
Gametocytes
Gels
Germ cells
Life Sciences
Male
Organ culture
Organ Culture Techniques
Original Article
Pachytene
Rats
Rats, Wistar
Sexual Maturation
Spermatogenesis
Spermatozoa
Testes
Testis
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Summary:Testicular tissue freezing before gonadotoxic treatments allows the preservation of fertility for children suffering from cancer. Recently, the testis organ culture method was presented as a relevant method to restore the fertility of these patients. However, the yield of spermatozoa production is low in the mouse model and no gamete has been obtained in vitro in the rat model. Here, we assess different cryopreservation protocols and culture conditions to improve the efficiency of in vitro maturation of rat prepubertal testes. Testes from male rats aged 5 or 8 days post - partum were cultured onto agarose gels of different percentages. After determining the best culture conditions, different cryopreservation protocols were assessed. Finally, testicular tissues were cultured with media of various compositions and analyzed at different time points. Our results show that the cryopreservation protocols allow the preservation of tissue architecture, cell proliferation, with no or moderate increase of cell death. In vitro spermatogenesis did not proceed beyond the pachytene spermatocyte stage. Only 2 of the 6 tested media allowed the survival of differentiated germ cells over the 45-day culture period. In conclusion, this study highlights the necessity to further improve the organ culture method before applying it into the clinics.
ISSN:0090-6964
1573-9686
DOI:10.1007/s10439-020-02535-8