Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Anatomical Sciences & Cognitive Neuroscience, Faculty of Medicine, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.
Cell Tissue Res. 2020 Mar;379(3):577-587. doi: 10.1007/s00441-019-03092-w. Epub 2019 Sep 7.
The current study evaluates potential applications of Sertoli cell (SC)-conditioned medium (CM) and explores the effects of the conditioned medium on the spermatogenesis process in azoospermic mice. For this study, 40 adult mice (28-30 g) were divided into 4 experimental groups: (1) control, (2) DMSO 2% (10 μl), (3) busulfan (40 mg/kg single dose) and (4) busulfan/CM (10 μl). SCs were isolated from 4-week-old mouse testes. After using anesthetics, 10 μl of CM was injected over 3-5 min into each testis and subsequently, sperm samples were collected from the tail of the epididymis. Afterward, the animals were euthanized and testis samples were taken for histopathology experiments and RNA extraction in order to examine the expression of c-kit, STRA8 and PCNA genes. The data showed that CM notably increased the total sperm count and the number of testicular cells, such as spermatogonia, primary spermatocytes, round spermatids, SCs and Leydig cells compared with the control, DMSO and busulfan groups. Furthermore, the results showed that expression of c-kit and STRA8 was significantly decreased in the busulfan and busulfan/SC groups at 8 weeks after the last injection (p < 0.001) but no significant difference was found for PCNA compared with the control and DMSO groups (p < 0.05). These findings suggest that the Sertoli cell-conditioned medium may be beneficial as a practical approach for therapeutic strategies in reproductive and regenerative medicine.
本研究评估了支持细胞（SC）条件培养基（CM）的潜在应用，并探讨了条件培养基对无精子症小鼠生精过程的影响。在这项研究中，将 40 只成年小鼠（28-30g）分为 4 个实验组：（1）对照组，（2）DMSO 2%（10μl），（3）白消安（40mg/kg 单次剂量）和（4）白消安/CM（10μl）。从 4 周龄的小鼠睾丸中分离出 SC。使用麻醉剂后，将 10μl CM 在 3-5 分钟内注入每个睾丸，然后从附睾尾部收集精子样本。之后，处死动物并取睾丸样本进行组织病理学实验和 RNA 提取，以检查 c-kit、STRA8 和 PCNA 基因的表达。结果表明，与对照组、DMSO 组和白消安组相比，CM 明显增加了总精子数和睾丸细胞数量，如精原细胞、初级精母细胞、圆形精子细胞、SC 和 Leydig 细胞。此外，结果显示，在最后一次注射后 8 周，白消安和白消安/SC 组中 c-kit 和 STRA8 的表达显著降低（p<0.001），但与对照组和 DMSO 组相比，PCNA 无显著差异（p<0.05）。这些发现表明，支持细胞条件培养基可能作为生殖和再生医学中治疗策略的一种实用方法是有益的。
