Spermatogenesis Recovery Potentials after Transplantation of Adipose TissueDerived Mesenchymal Stem Cells Cultured with Growth Factors in Experimental Azoospermic Mouse Models

Objective Approximately 1% of the male population suffers from obstructive or nonobstructive azoospermia. Previous in vitro studies have successfully differentiated mesenchymal stem cells (MSCs) into germ cells. Because of immune modulating features, safety, and simple isolation, adipose tissuederived MSCs (ATMSCs) are good candidates for such studies. However, low availability is the main limitation in using these cells. Different growth factors have been investigated to overcome this issue. In the present study, we aimed to comparatively assess the performance of ATMSCs cultured under the presence or absence of three different growth factors, epidermal growth factor (EGF), leukemia inhibitory factor (LIF) and glial cell linederived neurotrophic factor (GDNF), following transplantation in testicular torsiondetorsion mice Materials and Methods This was an experimental study in which ATMSCs were first isolated from male Naval Medical Research Institute (NMRI) mice. Then, the mice underwent testicular torsiondetorsion surgery and received bromodeoxyuridine (BrdU)labeled ATMSCs into the lumen of seminiferous tubules. The transplanted cells had been cultured in different conditioned media, containing the three growth factors and without them. The expression of germ cellspecific markers was evaluated with realtime polymerase chain reaction (PCR) and westernblot. Moreover, immunohistochemical staining was used to trace the labeled cells. Results The number of transplanted ATMSCs resided in the basement membrane of seminiferous tubules significantly increased after 8 weeks. The expression levels of Gcnf and Mvh genes in the transplanted testicles by ATMSCs cultured in the growth factorssupplemented medium was greater than those in the control group (P 0.001 and P 0.05, respectively). The expression levels of the cKit and Scp3 genes did not significantly differ from the control group. ConclusionOur findings showed that the use of EGF, LIF and GDNF to culture ATMSCs can be very helpful in terms of MSC survival and localization.