Leydig cells as a potential target of a drug based on mesenchymal stromal cell secretome in doxorubicin induced toxic damage of spermatogenesis

Background. Previously, it has been shown in animal models that the secretome of mesenchymal stromal cells (MSCs) can restore the spermatogonial stem cell niche. In a mouse model of doxorubicin-induced toxic damage to spermatogenesis, it was shown that the biodistribution of MSC secretome following administration under the tunica albuginea of the testis occurs mainly in the interstitium, which consists predominantly of Leydig cells. However, the effect of MSC secretome on Leydig cells under doxorubicin-induced damage remains unexplored.

Aim. To study the effect of MSC secretome on Leydig cells under doxorubicin-induced damage in vitro and in vivo.

Materials and Methods. Spermatogenesis damage was modeled in C57Bl/6 mice, which received intraperitoneal injections of doxorubicin at a total dose of 10 mg/kg. Three groups of animals were studied: intact, damaged, and MSC secretome-treated. For in vitro studies, doxorubicin (at concentrations ranging from 0.15 to 1.5 µM) and MSC secretome were added to Leydig cell cultures.

Results. Following doxorubicin-induced damage in vivo, mouse testis weight decreased and did not recover after MSC secretome injection. The interstitial area increased upon damage, while administration of MSC secretome tended to reduce (normalize) the interstitial area. At the same time, the number of Leydig cells did not change significantly. In in vitro studies, exposure of Leydig cells to doxorubicin induced dose-dependent cell death. Addition of MSC secretome to Leydig cells under damage conditions enhanced cell survival and testosterone secretion, resulting in an increase in total testosterone content in the Leydig cell culture medium.

Conclusion. MSC secretome can act on Leydig cells under doxorubicin-induced damage by increasing their number and secretory activity, which may contribute to interstitial recovery. These findings provide a rationale for further investigation of MSC secretome as a promising therapeutic agent for restoring testicular function.

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