NemaLife 芯片:一种基于微柱的微流控培养装置,经过优化可用于爬行秀丽隐杆线虫的衰老研究。
NemaLife chip: a micropillar-based microfluidic culture device optimized for aging studies in crawling C. elegans.
机构信息
Department of Chemical Engineering, Texas Tech University, Lubbock, TX, 79409, USA.
Department of Surgery, Texas Tech University Health Sciences Center, Lubbock, TX, 79409, USA.
出版信息
Sci Rep. 2020 Oct 1;10(1):16190. doi: 10.1038/s41598-020-73002-6.
In this study, we report a microfluidic device for the whole-life culture of the nematode Caenorhabditis elegans that allows the scoring of animal survival and health measures. This device referred to as the NemaLife chip features: (1) an optimized micropillar arena in which animals can crawl, (2) sieve channels that separate progeny and prevent the loss of adults from the arena during culture maintenance, and (3) ports that allow rapid accessibility for feeding the adult-only population and introducing reagents as needed. The pillar arena geometry was optimized to accommodate the growing body size during culture and emulate the body gait and locomotion of animals reared on agar. Likewise, feeding protocols were optimized to recapitulate longevity outcomes typical of standard plate growth. Key benefits of the NemaLife Chip include eliminating the need to perform repeated manual transfers of adults during survival assays, negating the need for progeny-blocking chemical interventions, and avoiding the swim-induced stress across lifespan in animals reared in liquid. We also show that the culture of animals in pillar-less microfluidic chambers reduces lifespan and introduces physiological stress by increasing the occurrence of age-related vulval integrity disorder. We validated our pillar-based device with longevity analyses of classical aging mutants (daf-2, age-1, eat-2, and daf-16) and animals subjected to RNAi knockdown of age-related genes (age-1 and daf-16). We also showed that healthspan measures such as pharyngeal pumping and tap-induced stimulated reversals can be scored across the lifespan in the NemaLife chip. Overall, the capacity to generate reliable lifespan and physiological data underscores the potential of the NemaLife chip to accelerate healthspan and lifespan investigations in C. elegans.
在这项研究中,我们报告了一种用于线虫秀丽隐杆线虫全生命周期培养的微流控设备,该设备可用于动物生存和健康指标的评分。该设备被称为 NemaLife 芯片,具有以下特点:(1)优化的微柱竞技场,动物可以在其中爬行;(2)筛状通道,可分离后代并防止在培养过程中成年动物从竞技场中丢失;(3)端口,可快速接触到仅成年的群体并按需引入试剂。柱形竞技场的几何形状经过优化,可适应培养过程中身体尺寸的增长,并模拟在琼脂上饲养的动物的身体步态和运动。同样,喂养方案经过优化,可再现与标准平板生长典型的寿命结果。NemaLife 芯片的主要优势包括消除了在生存分析中反复手动转移成年动物的需要,无需进行阻止后代的化学干预,以及避免了在液体中饲养的动物在整个生命周期中的游泳诱导应激。我们还表明,在没有微柱的微流控腔室中培养动物会通过增加与年龄相关的阴道完整性障碍的发生来缩短寿命并引起生理应激。我们使用经典衰老突变体(daf-2、age-1、eat-2 和 daf-16)的寿命分析和与年龄相关基因(age-1 和 daf-16)的 RNAi 敲低动物验证了我们的基于微柱的设备。我们还表明,可以在 NemaLife 芯片中跨寿命评分咽部抽吸和敲击诱导的刺激反转等健康指标。总体而言,生成可靠的寿命和生理数据的能力突显了 NemaLife 芯片在秀丽隐杆线虫健康和寿命研究中的潜力。