Synthesis and characterization of phase-separated extracellular condensates in interactions with cells

Biomolecular condensates formed through liquid-liquid phase separation play key roles in intracellular organization and signaling, yet their function in extracellular environments remains largely unexplored. Here, we establish a model using heparan sulfate, a key component of the extracellular matrix, to study extracellular condensate-cell interactions. We demonstrate that heparan sulfate can form condensates with a positively charged counterpart in serum-containing solutions, mimicking the complexity of extracellular fluid, and supporting cell viability. We observe that these condensates adhere to cell membranes and remain stable, enabling a versatile platform for examining extracellular condensate dynamics and quantifying their rheological properties as well as their adhesion forces with cellular surfaces. Our findings and methodology open new avenues for understanding the organizational roles of condensates beyond cellular boundaries. • The article presents the first ever engineering extracellular condensate model. • The first use of SPM in cell-condensate interaction analysis is presented. • Distinctive condensation behaviors of heparan sulfate and heparin are identified.