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Strains matter: Success of murine in vitro spermatogenesis is dependent on genetic background
Joana M D Portela1, Callista L Mulder1, Saskia K M van Daalen1
1Center for Reproductive Medicine, Amsterdam UMC, Amsterdam Reproduction and Development Research Institute, University of Amsterdam, Meibergdreef 9, 1105 AZ, Amsterdam, the Netherlands.
Abstract:
The current strategy to preserve fertility of male prepubertal cancer patients consists of cryopreservation of a testicular tissue biopsy containing spermatogonial stem cells (SSCs). While in humans, fertility restoration strategies from prepubertal testicular tissues are still under investigation and have not yet resulted in complete germ cell differentiation, in mice various studies have described production of sperm and offspring through testicular organ culture and transplantation of in vitro propagated SSCs. Organ culture has shown to be successful in generating mature spermatozoa when using testicular fragments from various mouse strains, including CD1 and C57BL/6 J. Conversely, in vitro proliferation of SSCs from C57BL/6 J mice is highly inefficient when compared to other strains such as DBA2 or hybrid mice of C57BL/6 J and DBA2 with 75% C57BL/6 J background (B6D2F2). In this study, we investigated in vitro spermatogenesis by organ culture using testicular tissue from C57BL/6 J and B6D2F2 mice. Whereas spermatogenesis was initiated and completed in C57BL/6 J fragments, it could not be effectively supported in B6D2F2 testicular tissue. While maturation of Sertoli cells and Leydig cells functionality appeared to be identical between the two strains, in B6D2F2 tissue spermatogenesis did not proceed past the spermatocyte step, followed by a rapid decline of the number of all germ cells in the fragments. This suggests that the spermatogenic potential in vitro is dependent on specialized sites in the genome and therefore the organ culture conditions suboptimal for some strains of mice.
Insights
In vitro organ culture successfully supported spermatogenesis in C57BL/6J mouse testicular tissue. However, B6D2F2 mouse tissue showed limited germ cell development, indicating strain-specific limitations for fertility restoration research.
Area of Science:
- Reproductive Biology
- Developmental Biology
- Stem Cell Science
Background:
- Current fertility preservation for prepubertal cancer patients involves cryopreserving testicular tissue with spermatogonial stem cells (SSCs).
- While human applications are under investigation, mouse models show promise for fertility restoration via testicular organ culture and SSC transplantation.
- Organ culture has successfully produced sperm from various mouse strains, but SSC proliferation varies significantly between strains.
Purpose of the Study:
- To investigate the efficacy of in vitro testicular organ culture for supporting spermatogenesis in C57BL/6J and B6D2F2 mouse strains.
- To identify potential strain-specific factors influencing in vitro spermatogenesis and germ cell development.
Main Methods:
- Testicular tissue fragments from C57BL/6J and B6D2F2 mice were cultured in vitro.
- Spermatogenesis progression, germ cell numbers, and Sertoli/Leydig cell function were assessed.
Main Results:
- Complete spermatogenesis was achieved in C57BL/6J testicular fragments.
- In B6D2F2 tissue, spermatogenesis arrested at the spermatocyte stage, with a subsequent decline in germ cell populations.
- Sertoli and Leydig cell maturation appeared comparable between the two strains.
Conclusions:
- In vitro organ culture supports spermatogenesis effectively in C57BL/6J mice but not in B6D2F2 mice.
- The success of in vitro spermatogenesis is influenced by the mouse strain, suggesting genetic factors impact germ cell development in organ culture.
- Optimizing organ culture conditions may be necessary for specific mouse strains to advance fertility restoration strategies.
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