Cacciottola Luciana, Courtoy Guillaume E, Nguyen Thu Y T, Hossay Camille, Donnez Jacques, Dolmans Marie-Madeleine
Gynecology Research Unit, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Av. Mounier 52, bte B1.52.02, 1200, Brussels, Belgium.
IREC Imaging Platform, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Av. Hippocrate 55, 1200, Brussels, Belgium.
J Assist Reprod Genet. 2021 Jan;38(1):151-161. doi: 10.1007/s10815-020-02005-z. Epub 2020 Nov 12.
To investigate whether adipose tissue-derived stem cells (ASCs) protect the primordial follicle pool, not only by decreasing direct follicle loss but also by modulating follicle activation pathways.
Twenty nude mice were grafted with frozen-thawed human ovarian tissue from 5 patients. Ten mice underwent standard ovarian tissue transplantation (OT group), while the remaining ten were transplanted with ASCs and ovarian tissue (2-step/ASCs+OT group). Ovarian grafts were retrieved on days 3 (n = 5) and 10 (n = 5). Analyses included histology (follicle count and classification), immunohistochemistry (c-caspase-3 for apoptosis and LC3B for autophagy), and immunofluorescence (FOXO1 for PI3K/Akt activation and YAP for Hippo pathway disruption). Subcellular localization was determined in primordial follicles on high-resolution images using structured illumination microscopy.
The ASCs+OT group showed significantly higher follicle density than the OT group (p = 0.01). Significantly increased follicle atresia (p < 0.001) and apoptosis (p = 0.001) were observed only in the OT group. In primordial follicles, there was a significant shift in FOXO1 to a cytoplasmic localization in the OT group on days 3 (p = 0.01) and 10 (p = 0.03), indicating follicle activation, while the ASCs+OT group and non-grafted controls maintained nuclear localization, indicating quiescence. Hippo pathway disruption was encountered in primordial follicles irrespective of transplantation, with nuclear YAP localized in their granulosa cells.
We demonstrate that ASCs exert positive effects on the ovarian reserve, not only by protecting primordial follicles from direct death but also by maintaining their quiescence through modulation of the PI3K/Akt pathway.
研究脂肪组织来源的干细胞（ASC）是否不仅通过减少直接的卵泡损失，还通过调节卵泡激活途径来保护原始卵泡库。
将来自5名患者的冻融人卵巢组织移植到20只裸鼠体内。10只小鼠接受标准卵巢组织移植（OT组），其余10只移植ASC和卵巢组织（两步法/ASC + OT组）。在第3天（n = 5）和第10天（n = 5）取出卵巢移植物。分析包括组织学（卵泡计数和分类）、免疫组织化学（用于凋亡的c-caspase-3和用于自噬的LC3B）以及免疫荧光（用于PI3K/Akt激活的FOXO1和用于Hippo途径破坏的YAP）。使用结构照明显微镜在高分辨率图像上确定原始卵泡中的亚细胞定位。
ASC + OT组的卵泡密度显著高于OT组（p = 0.01）。仅在OT组中观察到卵泡闭锁（p < 0.001）和凋亡显著增加（p = 0.001）。在原始卵泡中，OT组在第3天（p = 0.01）和第10天（p = 0.03）FOXO1向细胞质定位有显著变化，表明卵泡激活，而ASC + OT组和未移植对照组保持核定位，表明静止。无论移植情况如何，在原始卵泡中均发现Hippo途径破坏，其颗粒细胞中有核YAP定位。
我们证明ASC对卵巢储备发挥积极作用，不仅通过保护原始卵泡免于直接死亡，还通过调节PI3K/Akt途径维持其静止状态。
