Towards A Synthetic Cell Model That Displays Receptor Mediated Endocytosis

Abstract

This project is an iterative exploration towards a minimal synthetic cell model based on an amphiphilic glycopolymer that assembles into GUVs that display receptor mediated endocytosis.
Chapter one is a general introduction chapter on the literature related to the present work including topics such as synthetic vesicles, natural cell membrane and its synthetic models, and the importance of transmembrane transport in the living cells.
Chapter two includes electroformation method applications on model molecules: lipid 1, 2 - dioleoyl - sn - glycerol - 3 - phosphocholine and the block copolymer poly (butadiene - b - ethylene oxide). Electroformation studies performed with a custom - made electroformation kit were accomplished successfully. Conditions suitable for the production of giant liposomes and polymersomes were obtained and the created structures were characterised, and analysed. A Full Factorial Design of Experiment Approach applied to the electroformation experiments on block copolymer poly (butadiene - b - ethylene oxide) revealed that the most influential factor on the final self - assembly outcome is the volume of deposited amphiphilic molecule on the glass slide.
Chapter three deals with giant unilamellar vesicle electroformation from the novel glycopolymer polyethylene - block - poly(ethylene glycol) β - D - glucoside and a variety of poly [N - 2 - (β - D - glucosyloxy) ethyl acrylamide] - b - (n-butylacrylate) glycopolymers with different block ratios and molar mass. Self - assembly experiments with novel amphiphilic materials were accomplished with desirable results - conditions
required for giant unilamellar vesicle formation were obtained; moreover, the collected data indicated that glycosylated block copolymer poly [N - 2 - (β - D - glucosyloxy) ethyl acrylamide] - b - (n-butylacrylate) with a block ratio 1 : 10 (hydrophilic block to hydrophobic block ratio) is a suitable amphiphilic material for the synthetic cell model creation.
Chapter four describes thorough studies performed on giant glycopolymersomes electroformed from polymer poly [N - 2 - (β - D - glucosyloxy) ethyl acrylamide] - b - (n-butylacrylate) with a block ratio 1 : 10. Polymersomes were reported to be stable and resistant to minor changes of osmolality and pH in their surrounding environment. Interaction studies confirmed that, despite the reported selective interactions between GUVs and PS - Con A beads, an evident uptake of nanoparticles in the glycopolymersome were not observed; however, findings presented in this chapter are promising and convincing that RME could be performed in a purely synthetic system in the near future upon applying desirable membrane modifications allowing the regulation of toughness and permeability.
Chapter five presents a recap of conclusions and general suggestions for continuation and further development of the project.
Chapter six describes general experimental procedures, materials and instrumentation utilised in this project.