Chen Shuwen, Wang Yanbin, Liao Liming, Meng Li, Li Juanjuan, Shi Cheng, Han Hongjing, Zheng Xiaofeng, Shen Huan
Reproductive Medical Center, Peking University People's Hospital, Peking University, Beijing 100044, China.
School of Life Sciences, Peking University, Beijing 100191, China.
Stem Cells Int. 2020 Dec 3;2020:8861557. doi: 10.1155/2020/8861557. eCollection 2020.
This study is aimed at investigating the repairing effect of mesenchymal stem cells and their exosomes from different sources on ovarian granulosa cells damaged by chemotherapy drugs-phosphoramide mustard (PM).
In this study, we choose bone marrow mesenchymal stem cells (BMSCs) and human placental mesenchymal stem cells (HPMSCs) for research. Then, they were cocultured with human ovarian granulosa cells (SVOG) injured by phosphoramide mustard (PM), respectively. -Galactosidase staining, flow cytometry, and Western blot were used to detect the changes in the senescence and apoptosis of SVOG cells before and after their coculture with the above two types of MSCs. Subsequently, exosomes from these two types of MSCs were extracted and added to the culture medium of SVOG cells after PM injury to test whether these two types of exosomes played a role similar to that of MSCs in repairing damaged SVOG cells.
PM treatment-induced apoptotic SVOG cells were significantly decreased after HPMSCs and BMSCs as compared with control group. After coculturing with these two types of MSCs, PM-treated SVOG cells showed significantly reduced senescence and apoptosis proportions as well as cleaved-Caspase 3 expression, and HPMSCs played a slightly stronger role than BMSCs in repairing SVOG cells in terms of the above three indicators. In addition, the ratios of senescent and apoptotic SVOG cells were also significantly reduced by the two types of exosomes, which played a role similar to that of MSCs in repairing cell damages.
The results indicated that BMSCs, HPMSCs, and their exosomes all exerted a certain repair effect on SVOG cells damaged by PM, and consistent repair effect was observed between exosomes and MSCs. The repair effect of exosomes secreted from BMSCs and HPMSCs on the SVOG cells was studied for the first time, and the results fully demonstrated that exosomes are the key carriers for MSCs to play their role.
本研究旨在探讨不同来源的间充质干细胞及其外泌体对化疗药物磷酰胺芥（PM）损伤的卵巢颗粒细胞的修复作用。
本研究选取骨髓间充质干细胞（BMSCs）和人胎盘间充质干细胞（HPMSCs）进行研究。然后，将它们分别与人卵巢颗粒细胞（SVOG）共培养，这些卵巢颗粒细胞已被磷酰胺芥（PM）损伤。采用β-半乳糖苷酶染色、流式细胞术和蛋白质免疫印迹法检测SVOG细胞与上述两种间充质干细胞共培养前后衰老和凋亡的变化。随后，提取这两种间充质干细胞的外泌体，并将其添加到PM损伤后的SVOG细胞培养基中，以测试这两种外泌体在修复受损SVOG细胞方面是否发挥与间充质干细胞类似的作用。
与对照组相比，HPMSCs和BMSCs共培养后，PM处理诱导的凋亡SVOG细胞显著减少。与这两种间充质干细胞共培养后，PM处理的SVOG细胞的衰老和凋亡比例以及裂解的Caspase 3表达显著降低，就上述三个指标而言，HPMSCs在修复SVOG细胞方面的作用略强于BMSCs。此外，两种外泌体也显著降低了SVOG细胞的衰老和凋亡比例，其在修复细胞损伤方面发挥了与间充质干细胞类似的作用。
结果表明，BMSCs、HPMSCs及其外泌体对PM损伤的SVOG细胞均有一定的修复作用，外泌体与间充质干细胞之间观察到一致的修复效果。首次研究了BMSCs和HPMSCs分泌的外泌体对SVOG细胞的修复作用，结果充分证明外泌体是间充质干细胞发挥作用的关键载体。
