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Background  Spermatogonial stem cell transplantation (SSCT) could become a fertility restoration tool for childhood cancer survivors. However, since in mice, the colonization efficiency of transplanted spermatogonial stem cells (SSCs) is only 12%, the efficiency of the procedure needs to be improved before clinical implementation is possible. Co-transplantation of mesenchymal stem cells (MSCs) might increase colonization efficiency of SSCs by restoring the SSC niche after gonadotoxic treatment.    Methods  A mouse model for long-term infertility was developed and used to transplant SSCs (SSCT,  n  = 10), MSCs (MSCT,  n  = 10), a combination of SSCs and MSCs (MS-SSCT,  n  = 10), or a combination of SSCs and TGFß1-treated MSCs (MSi-SSCT,  n  = 10).    Results  The best model for transplantation was obtained after intraperitoneal injection of busulfan (40 mg/kg body weight) at 4 weeks followed by CdCl 2  (2 mg/kg body weight) at 8 weeks of age and transplantation at 11 weeks of age. Three months after transplantation, spermatogenesis resumed with a significantly better tubular fertility index (TFI) in all transplanted groups compared to non-transplanted controls ( P  < 0.001). TFI after MSi-SSCT (83.3 ± 19.5%) was significantly higher compared to MS-SSCT (71.5 ± 21.7%,  P  = 0.036) but did not differ statistically compared to SSCT (78.2 ± 12.5%). In contrast, TFI after MSCT (50.2 ± 22.5%) was significantly lower compared to SSCT ( P  < 0.001). Interestingly, donor-derived TFI was found to be significantly improved after MSi-SSCT (18.8 ± 8.0%) compared to SSCT (1.9 ± 1.1%;  P  < 0.001), MSCT (0.0 ± 0.0%;  P  < 0.001), and MS-SSCT (3.4 ± 1.9%;  P  < 0.001). While analyses showed that both native and TGFß1-treated MSCs maintained characteristics of MSCs, the latter showed less migratory characteristics and was not detected in other organs.    Conclusion  Co-transplanting SSCs and TGFß1-treated MSCs significantly improves the recovery of endogenous SSCs and increases the homing efficiency of transplanted SSCs. This procedure could become an efficient method to treat infertility in a clinical setup, once the safety of the technique has been proven.    Electronic supplementary material  The online version of this article (10.1186/s13287-018-1065-0) contains supplementary material, which is available to authorized users.

Co-transplantation of mesenchymal stem cells improves spermatogonial stem cell transplantation efficiency in mice