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聚二甲基硅氧烷(PDMS)基底的表面功能化通过阻止非选择性吸附来促进胚胎培养。

Surface functionalization of poly(dimethylsiloxane) substrates facilitates culture of pre-implantation mouse embryos by blocking non-selective adsorption.

机构信息

Department of Mechanical and Industrial Engineering, University of Massachusetts, Amherst, MA 01003, USA.

Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA 01003, USA.

出版信息

J R Soc Interface. 2022 Apr;19(189):20210929. doi: 10.1098/rsif.2021.0929. Epub 2022 Apr 6.

Abstract

Poly(dimethylsiloxane) (PDMS) is widely used in biomedical settings such as microfluidics for its optical transparency, castability, gas permeability and relative biocompatibility. While PDMS devices with certain modifications or treatments have been used for mammalian pre-implantation embryo culture, it is unclear why native PDMS leads to significant embryo death. In this study, we employ Nile Red as a model hydrophobic small molecule to demonstrate that significant hydrophobic sequestration occurs on native PDMS substrates even with a bovine serum albumin-containing KSOM pre-equilibration. Our results suggest that this small molecule sequestration has detrimental effects on mouse embryo development in PDMS static culture wells, with 0% blastocyst development rates from embryos cultured on native PDMS. We found that prior saturation of the PDMS culture well with water vapour only rescues about 10% of blastocyst development rates, indicating osmolality alone is not responsible for the high rates of embryo arrest. We also present a safe and simple Pluronic F127 pretreatment for PDMS substrates that successfully circumvented the harmful effects of native PDMS, achieving a blastocyst and implantation rate akin to that of our polystyrene controls. Our results call into question how researchers and clinicians can account for the alterations in medium composition and embryo secretions when using hydrophobic substrates, especially in the mammalian embryo culture setting where minimum effective concentrations of peptides and amino acids are commonplace.

摘要

聚二甲基硅氧烷(PDMS)由于其光学透明性、可铸造性、气体透过性和相对生物相容性,在生物医学领域,如微流控领域得到了广泛的应用。虽然经过某些修饰或处理的 PDMS 器件已用于哺乳动物植入前胚胎培养,但尚不清楚为什么天然 PDMS 会导致胚胎大量死亡。在这项研究中,我们使用尼罗红作为模型疏水分子,证明即使在含有牛血清白蛋白的 KSOM 预平衡的情况下,天然 PDMS 基底上也会发生明显的疏水捕获。我们的结果表明,这种小分子的捕获对 PDMS 静态培养孔中的小鼠胚胎发育有不利影响,在天然 PDMS 上培养的胚胎的囊胚发育率为 0%。我们发现,仅用水蒸气饱和 PDMS 培养孔只能挽救约 10%的囊胚发育率,这表明渗透压本身并不是胚胎阻滞率高的原因。我们还提出了一种安全简单的 PDMS 基底预处理方法,即用普朗尼克 F127 预处理,成功地避免了天然 PDMS 的有害影响,达到了与聚苯乙烯对照物相当的囊胚和着床率。我们的研究结果表明,当使用疏水性基底时,研究人员和临床医生如何解释培养基成分和胚胎分泌物的变化,尤其是在最小有效浓度的肽和氨基酸在哺乳动物胚胎培养中很常见的情况下。

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