Capsules with bacteria and fungi in distinct compartments: A platform for studying microbes from different kingdoms and their cross-communication

Recently, we have created 'artificial cells' with an architecture mimicking that of typical eukaryotic cells. Our design uses common biopolymers like alginate and chitosan to create multi-compartment capsules (MCCs) via oil-free microfluidics. MCCs (similar to 500 mu m in diameter) can be engineered with multiple inner compartments, each with a distinct payload. This mimics the distinct organelles in eukaryotic cells, each of which has unique properties. In this study, we encapsulate microbial cells from two distinct kingdoms - Pseudomonas aeruginosa (bacteria) and Candida albicans (fungi) - in the inner compartments of MCCs. The two microbes are commonly found in biofilms at sites of infection in humans. We first demonstrate that the MCC can serve as a simple platform to observe the comparative growth of the cells in real time. Unlike typical co-culture in solution or on agar plates, the cells can grow in their own compartments without direct physical contact. Moreover, the hydrogel matrix in the compartments mimics the three-dimensional (3-D) environment that cells naturally encounter during their growth. Small molecules added to the solution are shown to permeate through the capsule walls and affect cell growth: for example, cationic surfactants inhibit the fungi but not the bacteria. Conversely, low pH and kanamycin inhibit the bacteria but not the fungi. Also, when the bacteria are present in adjacent compartments, the fungal cells mostly stay in a yeast morphology, meaning as spheroidal cells. In contrast, in the absence of the bacteria, the fungi transition into hyphae, i.e., long multicellular filaments. The inhibition of this morphological switch in fungal cells is shown to be induced by signaling molecules (specifically, the quorum sensing autoinducer-1 or AI-1) secreted by the bacteria. Thus, the MCC platform can also be used to detect cross-kingdom signaling between the compartmentalized microbes.

Automated summary

In this study, 'artificial cells' with an architecture mimicking that of eukaryotic cells were created using common biopolymers. The cells were encapsulated in multi-compartment capsules and were used to observe the comparative growth of microbial cells from different kingdoms. The capsules also mimicked the 3-D environment cells naturally encounter during growth. The study showed that small molecules can permeate through the capsule walls and affect cell growth, and that bacteria can induce a morphological switch in adjacent fungal cells through signaling molecules.