Department of Veterinary Medicine, Faculty of Animal Science and Food Engineering, University of São Paulo, São Paulo, Pirassununga, Brazil.
Mol Reprod Dev. 2019 Aug;86(8):1067-1080. doi: 10.1002/mrd.23223. Epub 2019 Jun 13.
The biotechnology for in vitro embryo production is becoming increasingly popular, being applied to humans and domestic animals. Embryo development can be achieved with either 20% or 5% oxygen tension. The extracellular vesicles (EVs) are secreted by different cell types and carry bioactive materials. Our objective was to determine the secretion pattern and micro RNA (miRNA) contents of EVs released in the bovine embryo culture environment-embryo and cumulus cell monolayer-on Days 3 and 7 of in vitro culture under two different oxygen tensions: High (20%) and low (5%). The EVs were isolated from the medium and analyzed to determine size, concentration, and miRNA levels. EVs concentration in low oxygen tension increased on Day 3 and decreased on Day 7. Additionally, altered EV miRNAs derived from the embryo-cumulus culture medium were predicted to regulate survival and proliferation-related pathways on Days 3 and 7. Moreover, miR-210 levels decreased in EVs isolated from the culture medium under high oxygen tension suggesting that this miRNA can be used as a marker for normoxia since it is associated with low oxygen tension. In summary, this study provides knowledge of the oxygen tension effects on EVs release and content, and potentially, on cell-to-cell communication during in vitro bovine embryo production.
体外胚胎生产的生物技术越来越受欢迎，已应用于人类和家畜。胚胎发育可以在 20%或 5%的氧气张力下进行。细胞外囊泡 (EVs) 由不同类型的细胞分泌，携带生物活性物质。我们的目的是确定在体外培养的第 3 天和第 7 天，在两种不同的氧气张力（高氧 20%和低氧 5%）下，在牛胚胎培养环境（胚胎和卵丘细胞单层）中释放的 EVs 的分泌模式和 micro RNA (miRNA) 含量。从培养基中分离 EVs 并进行分析，以确定其大小、浓度和 miRNA 水平。低氧张力下的 EV 浓度在第 3 天增加，在第 7 天减少。此外，来自胚胎-卵丘培养物培养基的改变的 EV miRNAs 预计会在第 3 天和第 7 天调节与存活和增殖相关的途径。此外，在高氧张力下从培养基中分离的 EVs 中的 miR-210 水平降低，这表明该 miRNA 可以用作正常氧的标志物，因为它与低氧张力有关。总之，这项研究提供了有关氧气张力对 EVs 释放和含量的影响的知识，并且可能对体外牛胚胎生产期间的细胞间通讯有影响。
