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评估细胞外囊泡和培养基中的 gDNA 作为人类胚胎发育能力的可能指标。

Evaluation of extracellular vesicles and gDNA from culture medium as a possible indicator of developmental competence in human embryos.

机构信息

Universidad de Concepción, Chillán, Chile.

Centro de Reproducción Asistida y Especialidades de la Mujer (CRAM), Concepción, Chile.

出版信息

Zygote. 2021 Apr;29(2):138-149. doi: 10.1017/S0967199420000593. Epub 2020 Oct 29.

Abstract

Human embryos generated in vitro have a high incidence of chromosomal abnormalities that negatively affect pregnancy rate. Embryos generated in vitro secrete extracellular vesicles (EVs) into the culture medium that could be used potentially as indicators of embryo competence. This research aimed to evaluate the concentration and size of EVs and their gDNA content as an indicator of developmental competence in human embryos. Human embryos generated by intracytoplasmic sperm injection (ICSI) were classified morphologically as of either TOP, FAIR or POOR quality. Culture medium and developmentally arrested embryos (which were not able to be used for embryo transfer) were collected. Microvesicles, exosomes (MV/Exo) and apoptotic bodies (ABs) were isolated from culture medium. Nanoparticle tracking analysis (NTA) and array comparative genomic hybridization (aCGH) analysis were performed to evaluate EVs and their gDNA content. From NTA, the diameter (mean) of MVs/Exo from TOP quality embryos was higher (112.17 nm) compared with that of FAIR (108.02) and POOR quality embryos (102.78 nm) (P < 0.05). aCGH analysis indicated that MVs/Exo and ABs carried gDNA with the presence of 23 chromosome pairs. However, when arrested embryos were compared with their respective MVs/Exo and ABs, the latter had an increased rate of chromosomal abnormalities (24.9%) compared with embryos (8.7%) (P < 0.05). In conclusion, the size of EVs from culture medium might be an alternative for evaluating competence of human embryos, however more studies are needed to validate the use of gDNA from EVs as an indicator of embryo competence.

摘要

体外培养的人类胚胎染色体异常发生率较高,这对妊娠率有负面影响。体外培养的胚胎会向培养液中分泌细胞外囊泡(EVs),这些囊泡可能可以作为胚胎活力的潜在指标。本研究旨在评估 EVs 的浓度和大小及其 gDNA 含量,以评估人类胚胎的发育能力。通过胞浆内单精子注射(ICSI)生成的人类胚胎根据形态学特征分为 TOP、FAIR 或 POOR 质量等级。收集培养液和发育停滞的胚胎(不能用于胚胎移植)。从培养液中分离出微囊泡、外泌体(MV/Exo)和凋亡小体(AB)。通过纳米颗粒跟踪分析(NTA)和比较基因组杂交分析(aCGH)评估 EVs 及其 gDNA 含量。从 NTA 结果来看,TOP 质量胚胎的 MV/Exo 的直径(平均值)较高(112.17nm),与 FAIR(108.02nm)和 POOR 质量胚胎(102.78nm)相比(P<0.05)。aCGH 分析表明,MV/Exo 和 AB 携带 gDNA,存在 23 对染色体。然而,当将停滞胚胎与其各自的 MV/Exo 和 AB 进行比较时,后者的染色体异常率(24.9%)高于胚胎(8.7%)(P<0.05)。总之,培养液中 EVs 的大小可能是评估人类胚胎活力的替代指标,但需要进一步研究来验证 EVs 中的 gDNA 作为胚胎活力的指标的使用。

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