In this talk, I will present recent research results from our group and collaborators with whom we are developing novel techniques to produce complex cell-like membranes in artificialcells and exploring the minimal requirements of a membrane leading to a cell division process. I will first present a strategy recently developed in our group, based on stabilizing droplets with functionalized nanoparticles, to produce giant unilamellar vesicles (GUVs) with complex membranes. This new method enables the integration of a correctly-folded membrane protein (Band 3) as well as the possibility of tuning ist orientation and its distribution within raft-like lipid nanodomains. Then I will present a diversity of strategies we are investigating to modify and hold the shape of GUVs. Among the different strategies we are pursuing, I will present results on the use of hydrogels, self-assembling peptides, and 3D nano-printed structures. Finally, I will discuss our work on developing a cell membrane capable of supporting cell division. We are approaching this problem by the isolation of active components directly from “living cells” followed by their encapsulation into GUVs in order to maximally preserve the global functionality of the original cytoplasm while only modifying the membrane components. I will specifically illustrate this process with the work we are doing on cell
extracts from eggs of Xenopus laevis. These cells are first “demembranized” by stripping off the cytoplasm from other components, which is then “remembranized” artificially. I will show how this approach, at the cross-roads of top-down and bottom-up synthetic biology, can retain the activity of the cellular machinery responsible for cell division, including the formation of nuclear envelopes and their subsequent breakdown, the generation of contractile actomyosin rings, and, more interestingly, the ability to establish deformations on the membrane that, in some cases, can lead to a cell division-like process.

Jorik Waeterschoot[1], Willemien Gosselé[1], Hojjat Alizadeh Zeinabad[1], Jiangyu Gan[1], Špela Lemež[1] Shomdutta Roy[1], Sara Battista[2], Arántzazu González-Campo[2], Valentina Corvaglia[3], Fabrizio Gelain[3] and Xavier Casadevall i Solvas[1]; [1] KU Leuven, BELGIUM; [2] Institute of Materials
Science of Barcelona, SPAIN; [3] Center for nanomedicine and tissue engineering, ITALY