Objective

To determine whether fluorescence lifetime imaging microscopy (FLIM) detects differences in metabolic state among cumulus cell samples and whether their metabolic state is associated with patient age, body mass index (BMI), and antimüllerian hormone (AMH) level and maturity of the oocyte.

Design

Prospective observational study.

Setting

Academic laboratory.

Patient(s)

Cumulus cell (CC) clusters from cumulus-oocyte complexes were collected from patients undergoing assisted reproductive technology treatment after oocyte retrieval and vitrified.

Intervention(s)

Cumulus cell metabolism was assessed using FLIM to measure autofluorescence of nicotinamide adenine (phosphate) dinucleotide and flavine adenine dinucleotide, endogenous coenzymes essential for cellular respiration and glycolysis. Patient age, BMI, and AMH level and the maturity of the corresponding oocytes were recorded.

Main Outcome Measure(s)

Quantitative information from FLIM was obtained regarding metabolite concentrations from fluorescence intensity and metabolite enzyme engagement from fluorescence lifetimes. Associations were investigated between each FLIM parameter and oocyte maturity and patient age, BMI, and AMH. Variance between CC clusters within and between patients was determined.

Result(s)

Of 619 CC clusters from 193 patients, 90 were associated with immature oocytes and 505 with metaphase II oocytes. FLIM enabled quantitative measurements of the metabolic state of CC clusters. These parameters were significantly correlated with patient age and AMH independently, but not with BMI. Cumulus cell nicotinamide adenine (phosphate) dinucleotide FLIM parameters and redox ratio were significantly associated with maturity of the enclosed oocyte.

Conclusion(s)

FLIM detects variations in the metabolic state of CCs, showing a greater variance among clusters from each patient than between patients. Fluorescence lifetime imaging microscopy can detect CC metabolic associations with patient age and AMH and variations between mature and immature oocytes, suggesting the potential utility of this technique to help identify superior oocytes.

中文翻译：

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人类卵丘细胞的代谢成像揭示了卵丘细胞代谢特征、患者临床因素和卵母细胞成熟度之间的关联

客观的

确定荧光寿命成像显微镜 (FLIM) 是否检测到卵丘细胞样本之间代谢状态的差异，以及它们的代谢状态是否与患者年龄、体重指数 (BMI) 和抗苗勒管激素 (AMH) 水平和卵母细胞成熟度相关。

设计

前瞻性观察研究。

环境

学术实验室。

患者）

从卵母细胞取出和玻璃化后接受辅助生殖技术治疗的患者收集来自卵丘-卵母细胞复合体的卵丘细胞 (CC) 簇。

干预措施

使用 FLIM 评估积云细胞代谢，以测量烟酰胺腺嘌呤（磷酸盐）二核苷酸和黄素腺嘌呤二核苷酸（细胞呼吸和糖酵解所必需的内源性辅酶）的自发荧光。记录患者年龄、BMI 和 AMH 水平以及相应卵母细胞的成熟度。

主要观察指标）

从 FLIM 获得了关于来自荧光强度的代谢物浓度和来自荧光寿命的代谢物酶参与的定量信息。研究了每个 FLIM 参数与卵母细胞成熟度以及患者年龄、BMI 和 AMH 之间的关联。确定了患者内部和患者之间 CC 簇之间的差异。

结果）

在来自 193 名患者的 619 个 CC 簇中，90 个与未成熟卵母细胞相关，505 个与中期 II 卵母细胞相关。FLIM 能够定量测量 CC 簇的代谢状态。这些参数与患者年龄和 AMH 独立显着相关，但与 BMI 无关。卵丘细胞烟酰胺腺嘌呤（磷酸盐）二核苷酸 FLIM 参数和氧化还原比与封闭卵母细胞的成熟度显着相关。

结论

FLIM 检测 CC 代谢状态的变化，显示每个患者的簇之间的差异大于患者之间的差异。荧光寿命成像显微镜可以检测 CC 代谢与患者年龄和 AMH 之间的关联以及成熟和未成熟卵母细胞之间的差异，表明该技术在帮助识别优质卵母细胞方面的潜在用途。