Vanderbilt Biophotonics Center and Department of Biomedical Engineering, Vanderbilt University, Nashville, TN, 37232, USA.
Tennessee Fertility Institute, Franklin, TN, 37067, USA.
Sci Rep. 2024 Aug 26;14(1):19757. doi: 10.1038/s41598-024-70517-0.
Despite its high cost, the success rate for in vitro fertilization (IVF) remains < 33% in humans, driving the need for new techniques to improve embryo culture outcomes. The well-of-the-well (WOW) culture system is a platform for in-vitro mammalian embryo culture that has been shown to enhance the developmental competence of embryos and clinical pregnancy rates in humans. However, discovery and testing of the best design for optimal embryo culture quality is hindered by the lack of a method to flexibly produce WOW dishes of various designs. Here, we present a low-cost and simple method to fabricate WOW dishes with microwells of arbitrary shapes and dimensions. We use a low-cost 3D printing service to fabricate a poly(dimethylsiloxane) (PDMS)-based WOW insert that can be paired with a standard in vitro fertilization (IVF) dish to create WOW dishes with new microwell shapes, including pyramidal and hemispherical designs. We validate the fabrication quality of the WOW inserts and demonstrate the utility of the assembled WOW dishes for observation and grading of mouse embryo quality. Moreover, our results indicate that WOW dishes with hemispherical microwells result in better culture outcomes than traditional flat-bottomed IVF dishes and those with other microwell shapes, including the semi-elliptical microwells used in commercial WOW dishes. The proposed fabrication strategy thus provides a way to rapidly fabricate and test new WOW dishes that may bolster IVF success rates.
尽管体外受精 (IVF) 的成本很高，但成功率仍<33%，这促使人们需要新的技术来提高胚胎培养的效果。Well-of-the-well (WOW) 培养系统是一种体外哺乳动物胚胎培养平台，已被证明可以提高胚胎的发育能力和人类的临床妊娠率。然而，由于缺乏一种灵活制作各种设计的 WOW 培养皿的方法，因此限制了对最佳胚胎培养质量设计的发现和测试。在这里，我们提出了一种低成本且简单的方法来制造具有任意形状和尺寸微槽的 WOW 培养皿。我们使用低成本的 3D 打印服务来制造基于聚二甲基硅氧烷 (PDMS) 的 WOW 插件，该插件可以与标准的体外受精 (IVF) 培养皿配对，从而制造出具有新微槽形状的 WOW 培养皿，包括金字塔形和半球形设计。我们验证了 WOW 插件的制造质量，并展示了组装后的 WOW 培养皿在观察和分级小鼠胚胎质量方面的实用性。此外，我们的结果表明，具有半球形微槽的 WOW 培养皿比传统的平底 IVF 培养皿以及具有其他微槽形状（包括商业 WOW 培养皿中使用的半椭圆形微槽）的培养皿的培养效果更好。因此，所提出的制造策略为快速制造和测试新的 WOW 培养皿提供了一种方法，这可能会提高 IVF 的成功率。
