AggreBots: Configuring CiliaBots through guided, modular tissue aggregation

Ciliated biobots (CiliaBots) are engineered tissues capable of self-actuated propulsion via exterior motile cilia. While correlations have been observed between CiliaBot motility and morphology, direct control of morphological features to deliver desired motility outcomes remains unexplored. Here, we describe the engineering of aggregated CiliaBots (AggreBots) to augment control over CiliaBot structural parameters and, consequently, motility patterns through guided, modular aggregation of human airway epithelial spheroids [referred to as CiliaBot building blocks (CBBs)]. Multi-CBB aggregation generated rod-, triangle-, and diamond-shaped AggreBots, altering tissue geometry without sacrificing surface cilia density or inter-CBB boundary fidelity. The further introduction of CCDC39-mutated CBBs as cilia-inactive modules enabled the generation of hybrid AggreBots with precision modulation of active cilia distribution, further empowering alterations to motility patterns. Our results demonstrate the potential of AggreBots as living tissue propellers with morphological "levers" by which modifications to tissue motility can be theoretically planned and experimentally verified.