MicroRNAs in spent blastocyst culture medium are derived from trophectoderm cells and can be explored for human embryo reproductive competence assessment
MicroRNAs in spent blastocyst culture medium are derived from trophectoderm cells and can be explored for human embryo reproductive competence assessment
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DOI：
10.1016/j.fertnstert.2015.09.014
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发表时间：
2016-01-01
影响因子：
6.7
通讯作者：
Rienzi, Laura
中科院分区：
文献类型：
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作者：
Capalbo, Antonio;Ubaldi, Filippo Maria;Rienzi, Laura
目的：评估是否可以从用过的囊胚培养基 (SBM) 中准确分析细胞外 microRNA (miRNA) 并用作胚胎生物标志物。设计：前瞻性队列研究。设置：私人和学术体外受精中心。患者：来自五个优质人类囊胚的无内细胞团的滋养外胚层 (TE) 样本及其相对 SBM。干预措施：基于定量的 miRNA 纯化和分析方案聚合酶链反应在人胚胎干细胞 (hESC) 和具有或不具有生物变异性的 SBM 上进行验证。主要结果指标：分析培养基中与 TE 细胞相关的 miRNA，比较植入和未植入的整倍体囊胚之间的 miRNA 图谱。结果：收集来自卵裂期、桑椹胚期和囊胚期胚胎的培养基，以研究 miRNA 的存在。前瞻性地从整倍体植入 (n = 25) 和未植入的囊胚 (n = 28) 中收集 SBM 以进行比较。我们假设人类胚胎在培养基中分泌可用作生物标志物的 miRNA。 TE 和 SBM 样本的比较分析表明，SBM 中检测到的 miRNA 中有 96.6%（59 中的 57；95 CI，88.3-99.6）是由 TE 细胞表达的，表明 TE 起源。从卵裂和桑椹胚阶段胚胎收集的培养基显示出与空白相似的模式，表明从用过的培养基中进行的 miRNA 分析仅适用于囊胚。对整倍体植入和未植入囊胚的 SBM 进行 MicroRNA 分析，突出显示两种 miRNA（miR-20a、miR-30c），它们在前者中浓度增加，并且通过计算机预测参与 23 条植入相关途径。结论：可以以高重复性对培养基中人类囊胚分泌的 MicroRNA 进行分析，并且可以进一步探索该方法用于非侵入性胚胎选择。 (C) 2016 年，美国生殖医学会。
Objective: To assess whether extracellular microRNAs (miRNAs) can be accurately profiled from spent blastocyst culture media (SBM) and used as embryonic biomarkers.Design: Prospective cohort study.Setting: Private and academic in vitro fertilization centers.Patient(s): Inner cell mass-free trophectoderm (TE) samples and their relative SBM from five good-quality human blastocysts.Intervention(s): Protocol for miRNA purification and analysis based on quantitative polymerase chain reaction set and validated on human embryonic stem cells (hESCs) and on SBM with and without biological variability.Main Outcomes Measure(s): Analysis of miRNAs in culture media in relation with TE cells and comparison of miRNA profiles between implanted and unimplanted euploid blastocysts.Result(s): Culture media from embryos in the cleavage, morula, and blastocyst stages were collected to investigate the presence of miRNAs. The SBM were prospectively collected from euploid implanted (n = 25) and unimplanted blastocysts (n = 28) for comparison. We hypothesized that human embryos secrete miRNAs in culture media that can be used as biomarkers. The comparative analysis of TE and SBM samples revealed that 96.6% (57 of 59; 95 CI, 88.3-99.6) of the miRNAs detected in the SBM were expressed from TE cells, suggesting a TE origin. The culture media collected from cleavage and morula stage embryos showed a pattern similar to blanks, suggesting that miRNAs profiling from spent culture media applies only for blastocysts. MicroRNAs analysis of SBM from euploid implanted and unimplanted blastocysts highlighted two miRNAs (miR-20a, miR-30c) that showed increased concentrations in the former and were predicted in silico to be involved in 23 implantation-related pathways.Conclusion(s): MicroRNAs secreted from human blastocysts in culture media can be profiled with high reproducibility, and this approach can be further explored for noninvasive embryo selection. (C) 2016 by American Society for Reproductive Medicine.