Embryotools R&D Center, Parc Científic de Barcelona, Barcelona, Spain.
ART Institute of Washington, Hudson, NY, USA.
Hum Reprod. 2021 Feb 18;36(3):605-613. doi: 10.1093/humrep/deaa370.
Which lab-related factors impact the culture system's capacity to maintain a stable osmolality during human embryo culture?
Incubator humidity, the volume of mineral oil, the type of culture media and the design of time-lapse dishes have been identified as important parameters that can cause an impact on media evaporation and consequently osmolality during culture.
Culture medium is a critical component in human embryo culture. Minimizing its evaporation during culture is an adequate strategy to stabilize osmolality and, as a result, improving culture conditions and clinical outcomes.
STUDY DESIGN, SIZE, DURATION: The studied variables included media composition and supplementation; volume of mineral oil; incubator humidification; and the type of dish and incubator used. Additionally, six time-lapse dish models were compared in their ability to prevent evaporation.
PARTICIPANTS/MATERIALS, SETTING, METHODS: Dishes were incubated in parallel to analyze osmolality during culture between groups: synthetic oviductal medium enriched with potassium versus human tubal fluid medium; protein versus no protein supplementation; dry versus humid atmosphere; high versus low volume of mineral oil. Additionally, media evaporation was compared between six models of time-lapse dishes with distinct designs, cultured in a joint incubator. Two of them were retested in their corresponding incubator to analyze the dish-incubator fit. Daily osmolality measurements were compared between groups. Linear regression was performed to analyze evaporation rates.
Protein supplementation did not significantly affect evaporation. Contrarily, humidity levels inside the incubators, the volume of mineral oil and the type of culture media, played an important role in osmolality stabilization. The design of time-lapse dishes and their recommended preparation protocol heavily influenced their evaporation rates, which were further altered by each incubator's characteristics. Media with initially high osmolalities had a bigger risk of reaching hypertonic levels during culture.
LIMITATIONS, REASONS FOR CAUTION: While numerous, the studied variables are limited and therefore other factors could play a role in osmolality dynamics, as well. Incontrollable atmospheric factors could also result in some variation in the observed results between different centers and laboratories.
Published literature has extensively described how hypertonic media may impair embryo development and negatively affect clinical outcomes; therefore, maintaining a stable osmolality during culture should be considered essential. This work is of interest both for embryologists when analyzing their culture system and methodologies, as well as manufacturers in charge of designing IVF consumables.
STUDY FUNDING/COMPETING INTEREST(S): This study was privately funded.
N/A.
哪些与实验室相关的因素会影响胚胎培养过程中培养液的渗透压稳定性？
已经确定了一些重要参数，包括培养箱的湿度、矿物油的体积、培养液的类型以及延时培养皿的设计，这些因素可能会影响培养液的蒸发，从而影响渗透压。
培养液是胚胎培养的关键组成部分。在培养过程中尽量减少培养液的蒸发，是稳定渗透压的有效策略，进而改善培养条件和临床结局。
研究设计、规模、持续时间：研究变量包括培养液的成分和补充剂、矿物油的体积、培养箱的加湿、所使用的培养皿类型和培养箱。此外，还比较了六种延时培养皿模型防止蒸发的能力。
参与者/材料、设置、方法：将培养皿置于平行环境中，在不同组之间分析培养过程中的渗透压，分别为富含钾的合成输卵管培养液与人类输卵管液培养液、添加蛋白质与不添加蛋白质、干燥环境与湿润环境、高矿物油体积与低矿物油体积。此外，还比较了六种具有不同设计的延时培养皿模型之间的培养液蒸发情况，这些培养皿在一个联合培养箱中进行培养。其中两种培养皿在各自的培养箱中进行了重新测试，以分析培养皿与培养箱的适配性。对各组的每日渗透压进行比较。采用线性回归分析蒸发率。
蛋白质补充剂对蒸发没有显著影响。相反，培养箱内的湿度水平、矿物油的体积以及培养液的类型在渗透压稳定方面起着重要作用。延时培养皿的设计及其推荐的准备方案对其蒸发率有很大影响，而每个培养箱的特点进一步改变了蒸发率。初始渗透压较高的培养液在培养过程中更有可能达到高渗状态。
局限性、谨慎的原因：尽管研究了许多变量，但它们是有限的，因此其他因素也可能影响渗透压动力学。不可控的大气因素也可能导致不同中心和实验室之间的观察结果存在差异。
已有大量文献描述了高渗培养液如何损害胚胎发育并对临床结局产生负面影响；因此，在培养过程中保持渗透压稳定应被视为至关重要的因素。这项工作不仅对分析培养系统和方法的胚胎学家有意义，对负责设计体外受精耗材的制造商也有意义。
研究资金/利益冲突：本研究由私人资助。
无。
