Novel co-culture plate enables growth dynamic-based assessment of contact-independent microbial interactions

Abstract

Interactions between microbes are central to the dynamics of microbial communities. Understanding these interactions is essential for the characterization of communities, yet challenging to accomplish in practice. There are limited available tools for characterizing diffusion-mediated, contact-independent microbial interactions. A practical and widely implemented technique in such characterization involves the simultaneous co-culture of distinct bacterial species and subsequent analysis of relative abundance in the total population. However, distinguishing between species can be logistically challenging. In this paper, we present a low-cost, vertical membrane, co-culture plate to quantify contact-independent interactions between distinct bacterial populations in co-culture via real-time optical density measurements. These measurements can be used to facilitate the analysis of the interaction between microbes that are physically separated by a semipermeable membrane yet able to exchange diffusible molecules. We show that diffusion across the membrane occurs at a sufficient rate to enable effective interaction between physically separate cultures. Two bacterial species commonly found in the cystic fibrotic lung, Pseudomonas aeruginosa and Burkholderia cenocepacia, were co-cultured to demonstrate how this plate may be implemented to study microbial interactions. We have demonstrated that this novel co-culture device is able to reliably generate real-time measurements of optical density data that can be used to characterize interactions between microbial species.

Exosome Research and Co-culture Study

“…Cell behavior is often affected by material surfaces, and the influence of the filter material on the bottom of the upper chamber on behavior cannot be overlooked. Moreover, since it is necessary (A) Filter separation: for vertical connection, Transwell systems, 64,65) microfluidic systems, 51) bioreactor systems, 49,[66][67][68] and horizontal connection (this review, interactive co-culture plate, Ginreilab Inc., Japan). (B) Solid separation: cells in droplets, [69][70][71] microfluidic system.…”

“…Co-culture systems are able to provide a variety of techniques for cell engineering and they are considered particularly effective for drug discovery. 51) Although high-throughput systems are required for drug discovery, constructing such systems in animal models is difficult in terms of cost and efficiency. Thus, developing a co-culture system with high-throughput capabilities is desirable in the future.…”

“…The main motivation behind employing co-culture systems is to examine interactions between natural cells and microorganisms 49) and to develop novel cell engineering techniques using such interactions. 50,51) Various terms are used to refer to co-culture systems, including "coculture," "co-culture," "mixed culture," "dual culture," "co-cultivation," and "co-incubation." The number of times these terms were used in publications obtained through a search of the PubMed literature database (referred to below as "co-culture-related papers") is shown in Supplementary Table 1.…”

Mapping Gene-Microbe Interactions: Insights from Functional Genomics Co-culture Experiments betweenSaccharomyces cerevisiaeandPseudomonasspp

“…Indeed, DRS2 is annotated with response to pheromone triggering conjugation with cellular fusion (10). Further experiments would be needed to discriminate between these possibilities, such as co-culture in membrane-separated wells (40). Also, further interpretation of the effects observed would be possible by incorporating genomic and transcriptomic information about the bacterial competitors.…”

Bridging evolutionary game theory and metabolic models for predicting microbial metabolic interactions

“…Despite continual progresses in determining metabolite exchanges using experimental approach such as 9 spatially separated apparatus designs [21,22], isotope probing [23,24] and tracing [25] techniques, 16s RNA 10 analysis [26] and metabolome analysis [27], we still need governing principles to predict and understand these 11 metabolic interactions.…”