IVI Foundation, Valencia, Spain; Reproductive Medicine Research Group, Valencia, Spain.
Fertility Preservation Unit, Women's Health Area, La Fe University Hospital, Valencia, Spain.
Fertil Steril. 2020 Mar;113(3):609-617.e3. doi: 10.1016/j.fertnstert.2019.10.030.
To evaluate whether specific ovarian decortication techniques vary in promoting ovarian cortex cryopreservation and transplant outcomes.
Experimental design.
University hospital.
ANIMAL(S): Nonobese diabetic (NOD)/severe combined immunodeficiency (SCID) female mice.
INTERVENTION(S): Human ovarian biopsy samples allocated to one of the following decortication procedures: scratching with scalpel blade (B), cutting with microsurgical scissors (M), separation with slicer (S), or no-separation (control, C). Parallel, in vivo experiment: decortication techniques combined with slow freezing (SF) and vitrification (VT) before xenograft into immunodeficient mice.
MAIN OUTCOME MEASURE(S): Follicular counts, apoptosis, shear stress, Hippo pathway and inflammation. In vivo: recovered grafts analyzed for follicular counts, angiogenesis, proliferation, and fibrosis.
RESULT(S): There were no differences in follicular density or number of damaged follicles between the decortication techniques in the in vitro study. Nevertheless, the M samples showed statistically significantly increased stromal damage compared with the controls and S samples, and up-regulation of Hsp60 shear stress gene expression. Decortication by both M and S inhibited the Hippo pathway, promoting gene expression changes. In the 21-day xenograft, total follicular density statistically significantly decreased compared with the nongrafted controls in all groups. Nevertheless, no differences were observed between the decortication techniques. Ovarian stroma vascularization was increased in the vitrified samples, but among the slow-freezing samples, the B samples had the lowest microvessel density. The M decorticated xenografts had increased fibrosis.
CONCLUSION(S): Decortication with a slicer causes less damage to ovarian tissue than other commonly used methods although microsurgical scissors seem to preserve slightly increased follicular numbers. Nevertheless, blade decortication seems to be a reliable technique for maintaining acceptable follicular conditions without inducing serious stromal impairment.
评估特定的卵巢皮质剥除技术在促进卵巢皮质冷冻保存和移植结果方面是否存在差异。
实验设计。
大学医院。
非肥胖型糖尿病（NOD）/严重联合免疫缺陷（SCID）雌性小鼠。
将人类卵巢活检样本分配到以下一种去皮质程序中：手术刀刀片刮除（B）、显微手术剪刀切割（M）、切片机分离（S）或无分离（对照，C）。平行的体内实验：去皮质技术与慢速冷冻（SF）和玻璃化（VT）结合，然后移植到免疫缺陷小鼠体内。
卵泡计数、细胞凋亡、剪切力、 Hippo 通路和炎症。体内：分析移植后回收的移植物的卵泡计数、血管生成、增殖和纤维化。
在体外研究中，不同去皮质技术之间的卵泡密度或受损卵泡数量没有差异。然而，与对照组和 S 组相比，M 组的基质损伤具有统计学显著增加，且热休克蛋白 60 剪切力基因表达上调。M 和 S 的去皮质均抑制 Hippo 通路，促进基因表达变化。在 21 天的异种移植中，与未移植对照组相比，所有组的总卵泡密度均显著降低。然而，不同去皮质技术之间没有差异。玻璃化样本中卵巢基质血管化增加，但在慢速冷冻样本中，B 样本的微血管密度最低。M 去皮质的异种移植有增加的纤维化。
与其他常用方法相比，切片机去皮质对卵巢组织的损伤较小，尽管显微手术剪刀似乎略微保留了更多的卵泡数量。然而，刀片去皮质似乎是一种可靠的技术，可以在不引起严重基质损伤的情况下保持可接受的卵泡条件。
