Follicular loss and tissue degeneration are great challenges in ovarian tissue culture systems. Mesenchymal stem cells (MSC) secrete a cocktail of growth factors and cytokines which supports adjacent cells and tissues. The aim of the current study was to investigate the impact of human bone marrow (hBM)-MSC, as co-culture cells, on human follicular development in ovarian cortical tissue (OCT) culture. For this purpose, warmed OCT fragments were co-cultured with hBM-MSC for 8 days and compared to monocultured OCT. During the culture period, ovarian follicle survival and development in the OCT were evaluated using histological observation, follicular developmental-related genes expression, and estradiol production. Furthermore, cell proliferation and apoptosis were assessed. The results showed that there were no significant differences in conserved ovarian follicles with a normal morphology between the two groups. However, the percentage of developing follicles, as well as follicular developmental gene expression, significantly increased in the co-culture group compared to the monoculture group. On the other hand, compared with the monoculture group, the co-culture group demonstrated a significant increase in cell proliferation, indicated by Ki67 gene expression, as well as a dramatic decrease in apoptotic cell percentage, revealed by TUNEL assay. These findings indicated that co-culturing of hBM-MSC with OCT could improve follicular activation and early follicular development in human ovarian tissue culture systems.

中文翻译：

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通过与间充质干细胞共培养改善冷冻保存的人卵巢皮质组织的原位卵泡激活和早期发育

卵泡丢失和组织变性是卵巢组织培养系统中的巨大挑战。间充质干细胞 (MSC) 分泌生长因子和细胞因子的混合物，支持相邻细胞和组织。本研究的目的是研究人骨髓 (hBM)-MSC 作为共培养细胞对卵巢皮质组织 (OCT) 培养中人卵泡发育的影响。为此，将加热的 OCT 片段与 hBM-MSC 共培养 8 天，并与单培养的 OCT 进行比较。在培养期间，通过组织学观察、卵泡发育相关基因的表达和雌二醇的产生来评估 OCT 中卵巢卵泡的存活和发育。此外，评估细胞增殖和细胞凋亡。结果显示，两组间形态正常的保守卵泡无显着差异。然而，与单一培养组相比，共培养组中发育卵泡的百分比以及卵泡发育基因表达显着增加。另一方面，与单一培养组相比，共培养组表现出细胞增殖的显着增加，由 Ki67 基因表达表明，以及通过 TUNEL 测定显示的凋亡细胞百分比急剧下降。这些发现表明 hBM-MSC 与 OCT 的共培养可以改善人卵巢组织培养系统中的卵泡活化和早期卵泡发育。与单一培养组相比，共培养组的发育卵泡百分比以及卵泡发育基因表达显着增加。另一方面，与单一培养组相比，共培养组表现出细胞增殖的显着增加，由 Ki67 基因表达表明，以及通过 TUNEL 测定显示的凋亡细胞百分比急剧下降。这些发现表明 hBM-MSC 与 OCT 的共培养可以改善人卵巢组织培养系统中的卵泡活化和早期卵泡发育。与单一培养组相比，共培养组的发育卵泡百分比以及卵泡发育基因表达显着增加。另一方面，与单一培养组相比，共培养组表现出细胞增殖的显着增加，由 Ki67 基因表达表明，以及通过 TUNEL 测定显示的凋亡细胞百分比急剧下降。这些发现表明 hBM-MSC 与 OCT 的共培养可以改善人卵巢组织培养系统中的卵泡活化和早期卵泡发育。共培养组表现出细胞增殖的显着增加，由 Ki67 基因表达表明，以及凋亡细胞百分比的显着下降，由 TUNEL 测定显示。这些发现表明 hBM-MSC 与 OCT 的共培养可以改善人卵巢组织培养系统中的卵泡活化和早期卵泡发育。共培养组表现出细胞增殖的显着增加，由 Ki67 基因表达表明，以及凋亡细胞百分比的显着下降，由 TUNEL 测定显示。这些发现表明 hBM-MSC 与 OCT 的共培养可以改善人卵巢组织培养系统中的卵泡活化和早期卵泡发育。