Secretions released from mesenchymal stem cells improve spermatogenesis restoration of cytotoxic treatment with busulfan in azoospermia mice

Abstract

This study aimed at the efficacy of sequential treatment of bone marrow‐derived mesenchymal stem cell secretion for busulfan‐treated azoospermia in mice. The conditioned media (CM) was obtained from bone marrow mesenchymal stem cells (MSCs) or 293 cells. Chemically induced azoospermia mice received 200 μl MSC‐CM or 293‐CM twice a week intravenously for three consecutive weeks. The histological assessment of spermatogenic recovery quantifying the expression of meiosis‐associated genes, and Sertoli cell barrier functional factors were assessed. The characteristics of TM4 cells (Sertoli cell line) after pre‐incubation of MSC‐CM in vitro were also obtained. The MSC‐CM group had the most spermatogenic colonies among the three groups (p < .05), but no spermatids were seen. Expressions of the meiosis‐associated genes Dazl, Vasa, Miwi, Stra8, CyclinA1, Pgk2 and Scp3 in MSC‐CM testis were remarkably higher compared with 293‐CM and busulfan groups respectively (p < .05). The levels of Sertoli cell barrier functional factors, for example ICAM‐1 and N‐cadherin, were significantly increased during MSC‐CM treatment (p < .05). Moreover, pre‐incubation of MSC‐CM particularly accelerated the CD54 (ICAM‐1) and CD44 expressions of TM4 cells and promoted cell inherent adhesion. MSC‐CM treatment can significantly improve the short‐term restoration of spermatogonial structures of chemically induced azoospermia related to facilitating Sertoli cell adhesion integrity.

Potency Assays for Mesenchymal Stromal Cell Secretome-Based Products for Tissue Regeneration

“…In the murine model of busulfan-induced spermatogenesis injury, it was shown that the systemic administration of MSC secretome leads to the increased mRNA expression of N-cadherin and ICAM1, which are involved in the barrier function of Sertoli cells. Moreover, the MSC secretome-treated group in the study showed the best results in experimental wound healing by the TM4 Sertoli cell line in a scratch assay conducted in the early stages of observation [88]. This result was consistent with the hypothesis that MSC secretome could attract Sertoli cells to the injured niche from specific transient zones of the adult testes.…”

The Role of Stem Cells and Their Derived Extracellular Vesicles in Restoring Female and Male Fertility

“…SCs can also reduce mitochondrial apoptosis to restore fertility [236]. Cai et al reported that BM-MSCs restore fertility in cases of busulfaninduced azoospermia by releasing proteins, including angiogenic factors, antiapoptotic factors, chemokines, cytokines, and immunoregulators [240]. Furthermore, normal spermatogenesis was restored in the individuals with testicular toxicity using a conditioned medium derived from BM-MSCs [241] and exosomes derived from urine-derived SCs [242].…”