F Ghanimat - Department of Anatomical Sciences, Embryology and Histology,Tehran, Iran
M Faraji Sani - Department of Anatomical Sciences, Embryology and Histology,Tehran, Iran
MA Abdollahifar - Department of Anatomical Sciences, Embryology and Histology,Tehran, Iran
M Movahedi - Department of Anatomical Sciences, Embryology and Histology,Tehran, Iran

Background: Busulfan is a chemotherapeutic drug which can induce subfertility and even sterility and azoospermia in the males. Conditioned media (CM) which is a good source of growth factors, cytokines, various enzymes, and other soluble factors secreted by adipose tissue-derived mesenchymal stem cells (AT-MSCs) may be effective in testes exposed to busul-fan. So we aimed to evaluate the effects of AT-MSCs-CM on histopathologic changes of mouse testes exposed to busulfan according to stereological study.Materials and Methods: In this study, we utilized 40 adult male NMRI mice (28‒30 g). All the animals were divided into 5 experimental groups randomly: (1) control, (2) DMEM (inject-ed by DMEM into testes), (3) DMSO (injected by DMSO intra-peritoneally), (4) busulfan (BSU) (single intraperitoneal dose of 40 mg/kg busulfan), and (5) conditioned media (CM) [(single intraperitoneal dose of 40 mg/kg busulfan) + AT-MSCs-CM]. All mice were kept for 35 days following injection. Finally, sperm samples were collected from the tail of the epididymis, the animals were euthanized and testis samples were taken for histological and stereological studies.Results: BSU group showed a significant decrease in volumes of the testis, interstitium, and tubules compared to the control, DMEM, and DMSO groups (P<0.05), although there was no significant difference between BSU and CM groups (P> 0.05). In addition, the number of spermatogenic, and leydig cells de-creased in BSU group compared to the control, DMEM, and DMSO groups, while CM could significantly improve that pa-rameters in the CM group. Sperm count and motility, and also the total length of seminiferous tubules increased in CM group compared to the BSU group.Conclusion: AT-MSCs-CM showed regenerative character-istics on histopathologic changes of mouse testes exposed to busulfan. Considering less concern about CM application com-pared to the stem cells, it can be an alternative choice for thera-peutically and research purposes in the disturbed testes.

Busulfan, Addipose Tissue (AT), Conditioned Medium(CM), Mesenchymal Stem Cells(MSC), Spermatogen-esis