NemaLife chip: a micropillar-based microfluidic culture device optimized for aging studies in crawling C. elegans.
NemaLife chip: a micropillar-based microfluidic culture device optimized for aging studies in crawling C. elegans.
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DOI：
10.1038/s41598-020-73002-6
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发表时间：
2020-10-01
影响因子：
4.6
通讯作者：
Vanapalli SA
中科院分区：
文献类型：
--
作者：
Rahman M;Edwards H;Birze N;Gabrilska R;Rumbaugh KP;Blawzdziewicz J;Szewczyk NJ;Driscoll M;Vanapalli SA
在这项研究中，我们报告了一种用于线虫线虫终身培养的微流控装置，该装置可以对动物的生存和健康措施进行评分。这种被称为NemaLife芯片的设备具有以下特点：(1)优化的微柱竞技场，动物可以在其中爬行；(2)筛选通道，分离后代，防止培养维护期间竞技场中失去成虫；(3)端口，允许快速进入，用于喂养仅成虫种群并根据需要引入试剂。柱子竞技场的几何形状进行了优化，以适应培养过程中不断增长的体型，并模拟琼脂饲养动物的身体步态和运动。同样，喂养方案进行了优化，以概括标准平板生长的典型长寿结果。NemaLife芯片的主要优点包括：在存活分析过程中不需要对成年动物进行重复的人工转移，不需要对后代进行化学干预，避免了在液体饲养的动物的整个生命周期中由游泳引起的应激。我们还表明，在无柱微流体室中培养动物会减少寿命，并通过增加与年龄相关的外阴完整性障碍的发生而引入生理应激。我们通过经典衰老突变体（daf-2、age-1、eat-2和daf-16）和RNAi敲低年龄相关基因（age-1和daf-16）的动物的寿命分析验证了我们的柱式装置。我们还表明，在NemaLife芯片中，可以对咽部泵送和叩击诱发的刺激逆转等健康寿命测量进行评分。总的来说，生成可靠的寿命和生理数据的能力强调了NemaLife芯片在加速线虫健康寿命和寿命研究方面的潜力。
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.
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影响因子：
5.3
作者：
GANDHI, S;SANTELLI, J;RAOSANADI, D
通讯作者：
RAOSANADI, D
影响因子：
2.7
作者：
CHALFIE, M;SULSTON, J
通讯作者：
SULSTON, J
影响因子：
6.1
作者：
Ai X;Zhuo W;Liang Q;McGrath PT;Lu H
通讯作者：
Lu H
DOI：
10.1073/pnas.88.12.5360
发表时间：
1991-06-01
影响因子：
11.1
作者：
HARMAN, D
通讯作者：
HARMAN, D
影响因子：
56.9
作者：
CHALFIE, M;TU, Y;PRASHER, DC
通讯作者：
PRASHER, DC