Cryoprotective role of pentoxifylline on proliferation and differentiation of spermatogonial stem cell following transplantation into azoospermic torsion mouse model
Cryoprotective role of pentoxifylline on proliferation and differentiation of spermatogonial stem cell following transplantation into azoospermic torsion mouse model
mehrnoush malekzadeh,1mansour homayoun,2Tayebeh Rastegar,3,*
1. Department of Anatomy, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran 2. Department of Anatomy, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran 3. Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
Introduction: Preserving the spermatogonial stem cells (SSCs) in long periods of time during the treatment of male infertility using stem cell banking systems and transplantation is an important issue. Therefore, this study was conducted to develop an optimal cryopreservation protocol for SSCs using 10 mM pentoxifylline (PTX) as an antioxidant in basal freezing medium.
Methods: Testicular
torsion—a mouse model for long-term infertility—was used to transplant fresh SSCs (n = 6), fresh SSCs treated with PTX (n = 6), cryopreserved SSCs with basal freezing medium (n = 6), and cryopreserved SSCs treated with PTX (n = 6). Eight
weeks after germ cell transplantation, samples were assessed for proliferation, through evaluation of Ddx4 and Id4 markers, and differentiation via evaluation of C‐Kit and Sycp3, Tnp1, Tnp2, and Prm1 markers.
Results: According to morphological and flow
cytometry results, SSCs are able to form colonies and express Gfra1, Id4, α6‐integrin, and β1‐integrin markers. We found positive influence from PTX on proliferative and differentiative markers in SSCs transplanted to azoospermic mice. In the recipient testis, donor SSCs formed spermatogenic colonies and sperm.
Conclusion: Respecting these data, adding pentoxifylline is a
practical way to precisely cryopreserve germ cells enriched for SSCs in cryopreservation, and this procedure could become an efficient method to restore fertility in a clinical setup. However, more studies are needed to ensure its safety in the long term.