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Microdroplet-Enabled Highly Parallel Co-Cultivation of Microbial Communities

机译:启用微滴的微生物群落高度平行共培养

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摘要

Microbial interactions in natural microbiota are, in many cases, crucial for the sustenance of the communities, but the precise nature of these interactions remain largely unknown because of the inherent complexity and difficulties in laboratory cultivation. Conventional pure culture-oriented cultivation does not account for these interactions mediated by small molecules, which severely limits its utility in cultivating and studying “unculturable” microorganisms from synergistic communities. In this study, we developed a simple microfluidic device for highly parallel co-cultivation of symbiotic microbial communities and demonstrated its effectiveness in discovering synergistic interactions among microbes. Using aqueous micro-droplets dispersed in a continuous oil phase, the device could readily encapsulate and co-cultivate subsets of a community. A large number of droplets, up to ∼1,400 in a 10 mm×5 mm chamber, were generated with a frequency of 500 droplets/sec. A synthetic model system consisting of cross-feeding E. coli mutants was used to mimic compositions of symbionts and other microbes in natural microbial communities. Our device was able to detect a pair-wise symbiotic relationship when one partner accounted for as low as 1% of the total population or each symbiont was about 3% of the artificial community.
机译:在许多情况下,天然微生物群中的微生物相互作用对于维持社区至关重要,但是由于实验室培养固有的复杂性和困难性,这些相互作用的确切性质在很大程度上仍然未知。常规的纯培养导向培养无法解释小分子介导的这些相互作用,这严重限制了其在培养和研究来自协同群落的“不可培养”微生物中的效用。在这项研究中,我们开发了一种用于共生微生物群落高度平行共培养的简单微流控设备,并证明了其在发现微生物之间的协同相互作用中的有效性。使用分散在连续油相中的水性微滴,该设备可以轻松封装和共同培养群落的子集。在10 mm×5 mm的腔室中,以500个液滴/秒的频率产生了大量的液滴,最高可达1400个。由交叉饲养的大肠杆菌突变体组成的合成模型系统用于模拟天然微生物群落中共生菌和其他微生物的组成。当一个伴侣仅占总种群的1%或每个共生体约占人工群落的3%时,我们的设备就能检测出成对的共生关系。

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