Multi-layer encapsulation of plant protoplasts

We report here on an attempt to create an engineered structure which can reproduce the physical and mechanical environment of the land plant sporangium. This work is part of a broader effort to understand the stress mechanics of the land plant sporangium and its role in initiating reproductive development. A second purpose is to extend the range of experimental methods available for the study of the physical environment of plant cell growth and the mechanics of trans-cellular signaling in plant development. We describe an experimental protocol, based on the microfluidic encapsulation of living plant protoplasts in multilayered microbeads composed of tunable hydrogel materials whose mechanical properties can be modified to mimic the stress-mechanics of the living sporangium. Our results demonstrate the successful encapsulation of living plant protoplasts in dual-layered microbeads consisting of an inner layer of gelled agarose and an outer layer of cross-linked alginate/methacrylate.