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Cryopreservation of adult primate testes

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Abstract

For primates that may unexpectedly die, spermatozoa recovered from the epididymis can be used for assisted reproductive technologies (ART). However, in cases of unsuccessful epididymal sperm retrieval, adult testes are the only source of sperm. This study reports cryopreservation of testicular tissue of three adult primates (mandrill, chimpanzee, and marmoset). For successful cryopreservation, optimization of concentrations of commonly used cryoprotectant such as dimethyl sulfoxide (DMSO) and combinations of foetal bovine serum (FBS) and DMSO is essential. Testicular tissues were cryopreserved in 10 % DMSO (D10) with 80 % FBS (D10S80) or 20 % DMSO (D20) with 20 % FBS (D20S20). The cryopreserved testes were evaluated for efficacy of cryopreservation by assessing the viability and acrosome status of retrieved sperm, expression of spermatozoa- and spermatid-specific proteins (PRM2 and TNP1) by immunohistochemistry, and expression of pro- and anti-apoptosis- (BAX/BCL2 ratio), early apoptosis- (Annexin V), and cell proliferation-specific (PCNA) proteins by western blot analysis. The results suggest that mandrill testis can be effectively cryopreserved in D10S80, marmoset testis in D10S80 and D20S20, whereas chimpanzee testis in D20.

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Acknowledgments

We thank the authorities of Nehru Zoological Park, Hyderabad, India, for providing the testis samples. This work was supported by the Council for Scientific and Industrial Research (CSIR), Government of India (GOI) (grant number BSC0207), and Central Zoo Authority (CZA), GOI (grant number GAP0389).

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Authors and Affiliations

  1. Laboratory for the Conservation of Endangered Species, Centre for Cellular and Molecular Biology, Council for Scientific and Industrial Research, Uppal Road, Hyderabad, 500 007, India

    Lavanya Pothana, Naresh Kumar Venna, Lalitha Devi, Anju Singh, Ipsita Chatterjee & Sandeep Goel

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  1. Lavanya Pothana
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  2. Naresh Kumar Venna
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  3. Lalitha Devi
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  4. Anju Singh
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  5. Ipsita Chatterjee
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  6. Sandeep Goel
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Corresponding author

Correspondence to Sandeep Goel.

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Supplementary figure 1

Sections of normal adult human testis stained with (A) TNP1 and (B) PRM2 antibodies as positive control (GIF 224 kb)

High Resolution image (TIF 4225 kb)

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Pothana, L., Venna, N.K., Devi, L. et al. Cryopreservation of adult primate testes. Eur J Wildl Res 62, 619–626 (2016). https://doi.org/10.1007/s10344-016-1024-y

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  • DOI: https://doi.org/10.1007/s10344-016-1024-y

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