Microfluidic Hanging Droplet as a Programmable Platform for Mammalian Egg Vitrification

IF 6.1 2区工程技术 Q1 BIOCHEMICAL RESEARCH METHODS

Lab on a Chip Pub Date : 2024-10-02 DOI:10.1039/d4lc00428k

Haidong Feng, Georgios Katsikis, India Napier, Gong Du, Josh Lim, Joseph Doyle, Scott R. Manalis, Linda G Griffith

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引用次数: 0

Abstract

Egg (oocyte) vitrification is the dominant method for preserving fertility for women of reproductive age. However, the method is typically performed by hand, requiring precise (~0.1 to 10 µL) and time-sensitive (~1 sec) liquid exchange of cryoprotectants (CPA) around eggs as well as fine handling of eggs (~100 µm) for immersion into liquid nitrogen (LN2). Here, we developed a microfluidic platform for programmable vitrification. Our platform is based on a millimeter-sized hanging droplet inside which a given egg is suspended and subjected to liquid exchanges within seconds. After programmable exposures to CPA, the egg is extracted from the liquid-air interface of the droplet using a motorized fine-tip instrument and immersed into LN2 for vitrification. To benchmark our platform with the manual method, we vitrified over a hundred mouse eggs and found comparable percentages (~95%) for post-vitrification survivability. In addition, our platform performs real-time microscopy of the egg thereby enabling future studies where its morphology may be linked to functional outcomes. Our study contributes to the ongoing efforts to enhance the automation of embryology techniques towards broader applications in reproductive medicine both for clinical and research purposes.

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微流体悬挂液滴作为哺乳动物卵子玻璃化的可编程平台

卵子（卵母细胞）玻璃化是育龄妇女保留生育能力的主要方法。然而，该方法通常由人工操作，需要精确（约 0.1 至 10 µL）、高时效（约 1 秒）地对卵子周围的冷冻保护剂（CPA）进行液体交换，并对卵子（约 100 µm）进行精细处理，以便浸入液氮（LN2）中。在此，我们开发了一种用于可编程玻璃化的微流体平台。我们的平台基于一个毫米大小的悬挂液滴，特定的卵子悬浮在液滴中，并在几秒钟内进行液体交换。在对 CPA 进行可编程暴露后，使用电动细尖仪器将卵子从液滴的液气界面中提取出来，然后浸入 LN2 进行玻璃化。为了将我们的平台与人工方法进行比较，我们对一百多个小鼠卵子进行了玻璃化处理，发现玻璃化后存活率相当（约 95%）。此外，我们的平台还能对卵子进行实时显微镜观察，从而在未来的研究中将卵子的形态与功能结果联系起来。我们的研究有助于不断提高胚胎学技术的自动化程度，从而在临床和研究目的的生殖医学中实现更广泛的应用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Lab on a Chip 工程技术-化学综合

CiteScore

11.10

自引率

8.20%

发文量

434

审稿时长

2.6 months

期刊介绍： Lab on a Chip is the premiere journal that publishes cutting-edge research in the field of miniaturization. By their very nature, microfluidic/nanofluidic/miniaturized systems are at the intersection of disciplines, spanning fundamental research to high-end application, which is reflected by the broad readership of the journal. Lab on a Chip publishes two types of papers on original research: full-length research papers and communications. Papers should demonstrate innovations, which can come from technical advancements or applications addressing pressing needs in globally important areas. The journal also publishes Comments, Reviews, and Perspectives.