Gynecology Research Unit, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium.
Society for Research into Infertility, Brussels, Belgium.
F S Sci. 2021 May;2(2):141-152. doi: 10.1016/j.xfss.2021.01.007. Epub 2021 Feb 5.
To investigate whether adipose tissue-derived stem cells (ASCs) modulate hypoxia and oxidative stress in human ovarian tissue transplants.
Prospective experimental study SETTING: Gynecological research unit in a university hospital PATIENT(S): Cryopreserved ovarian cortex from 5 adult women.
INTERVENTION(S): Thirty mice were grafted with frozen-thawed human ovarian tissue, with or without ASCs (2-step/ASCs+ovarian tissue [OT] group and OT group). The ovarian grafts were retrieved on days 3 (n = 5), 10 (n = 5), and 21 (n = 5). The 10 animals grafted for 21 days underwent in vivo evaluations using microdialysis. One piece of ovarian tissue per patient was fixed for analysis after thawing (non-grafted controls).
MAIN OUTCOME MEASURE(S): Direct reactive oxygen species were collected every second day after grafting by means of microdialysis. Analyses of ovarian fragments included immunolabeling for double CD34 (revascularization by host and graft components); immunofluorescence for hypoxia-inducible factor 1α (hypoxia-related response), nuclear factor erythroid 2-related factor 2 (oxidative stress-related response), and 8-hydroxy-deoxyguanosine (oxidative stress-related DNA damage); and gene expression (quantitative reverse transcription polymerase chain reaction) for vascular endothelial growth factor-A (neoangiogenesis), superoxide dismutase 2 (antioxidant activity), and nuclear respiratory factor 1 (mitochondrial biogenesis).
RESULT(S): Reactive oxygen species peaked earlier in the ASC group (day 2) compared with that in the OT group (day 10) after grafting. Total vascularization was stable in the ASC group at all time points, while it was lower in the OT group 3 days after grafting. Hypoxia-inducible factor 1α expression, also detected in non-grafted controls, was significantly lower in the ASC group than in the OT group on days 3 and 10. The increase in VEGF gene expression lasted significantly longer in the ASC group than in the OT group. There was no significant upturn in the oxidative stress-related response (nuclear factor erythroid 2-related factor 2 pathway) or oocyte DNA damage (8-hydroxy-deoxyguanosine) in any of the grafted groups.
CONCLUSION(S): Use of ASCs allows faster ovarian graft reperfusion and mitigates the hypoxia-related response through rapid revascularization, sustained by prolonged increase in vascular endothelial growth factor after grafting. No evidence of oxidative stress-related damage was detected irrespective of the transplantation strategy.
研究脂肪组织来源的干细胞（ASCs）是否调节人卵巢组织移植中的缺氧和氧化应激。
前瞻性实验研究
大学医院的妇科研究单位
5 名成年女性的冷冻卵巢皮质。
30 只小鼠接受冷冻-解冻的人卵巢组织移植，有无 ASC（2 步/ASC+卵巢组织[OT]组和 OT 组）。卵巢移植物在第 3 天（n=5）、第 10 天（n=5）和第 21 天（n=5）取出。接受 21 天移植的 10 只动物进行了体内微透析评估。每位患者解冻后的 1 块卵巢组织（非移植对照）用于分析。
移植物后每隔一天通过微透析收集直接活性氧。对卵巢组织片段的分析包括双重 CD34 免疫标记（宿主和移植物成分的再血管化）；缺氧诱导因子 1α（缺氧相关反应）、核因子红细胞 2 相关因子 2（氧化应激相关反应）和 8-羟基脱氧鸟苷（氧化应激相关 DNA 损伤）的免疫荧光；血管内皮生长因子-A（新生血管形成）、超氧化物歧化酶 2（抗氧化活性）和核呼吸因子 1（线粒体生物发生）的定量逆转录聚合酶链反应（qRT-PCR）。
与 OT 组（移植后第 10 天）相比，移植后 ASC 组（第 2 天）的活性氧峰值更早。在所有时间点，ASC 组的总血管化均保持稳定，而移植后第 3 天，OT 组的血管化水平较低。在 ASC 组中，缺氧诱导因子 1α的表达也在非移植对照组中检测到，在第 3 天和第 10 天明显低于 OT 组。ASC 组中 VEGF 基因表达的增加持续时间明显长于 OT 组。在任何移植组中，氧化应激相关反应（核因子红细胞 2 相关因子 2 途径）或卵母细胞 DNA 损伤（8-羟基脱氧鸟苷）均未明显增加。
使用 ASC 可加速卵巢移植物再灌注，并通过移植后血管内皮生长因子的持续增加来缓解快速再血管化引起的缺氧相关反应。无论移植策略如何，均未发现与氧化应激相关的损伤证据。
