Abstracts for tutorials on Wednesday May 18th
Microfluidics: an indispensable tool for building synthetic cells
Wilhelm Huck
Radboud University, The Netherlands
Building a synthetic cell inevitably will require bringing together a larger number of components into a small volume. Although many process will have to proceed via self-assembly of molecular components, the exquisite level of control over the manipulation of small volumes of liquids that microfluidics brings, opens up robust strategies for quantitative studies. In this talk, I will review some of the basic aspects of microfluidics, and highlight the use of both single phase and biphasic (droplet-based) microfluidic platforms in bottom-up synthetic cell projects.
Hacking the liposome formation
Orion Venero, Kate Adamala
University of Minnesota, United States of America
Liposomal encapsulation serves as the basis for the engineering of biomimetic and novel synthetic cells. Liposomes are normally formed using such methods as thin film rehydration (TFH), density mediated reverse emulsion encapsulation (REE), or one of many microfluidics-based approaches. While several microfluidics-based methods exist, capable of efficiently forming unilamellar liposomes, with uniform size and acceptable encapsulation rates, the main limitation associated with microfluidics is that trace amounts of carrier organic solvent remain present in the resultant membrane. A popular method bypasses the problem of residual solvent presence by first, evaporating all carrier solvent and thereby creating a thin lipid film used to prepare liposomes through subsequent methods. However, most protocols which utilize this methodology of thin film preparation are non-microfluidic protocols and thus do not produce uniform unilamellar liposomes.
DSCF, which stands for Droplet‐Shooting Centrifugal Formation, is a derivative liposome formation method that utilizes solvent-less lipid thin films to prepare highly uniform lipid bilayer membranes using a 3D printable microfluidics-system. In this way, DSCF methods avoid the solvent issues associated with microfuidics while producing liposomes with a high level of repeatability, similarly to microfluidics.
Various protocols for the preparation of DSCF liposomes exist whereby lipid-bilayer membranes are formed when lumen droplets pass through a lipids-in-oil solution and thence into an aqueous solution via centrifugal force. Utilizing this general DSCF mechanism it is possible to assemble highly uniform liposomes quickly and easily with tunable lumen and membrane chemistries.
Engineered soft interfaces in microfluidics: from the lab to industry
Jean-Christophe Baret
CNRS, Univ Bordeaux, CRPP, France
Droplet-based microfluidics has emerged over the past decade as a new technology for the miniaturization and automatisation of biological assays at a ultra-high throughput. More recently, it proved to be a powerful tool to design and create assembly lines for artificial cells, for both the construction of microcompartments scaffold and for the implementation of function within these compartments. The technology relies on microsystems for fluidic manipulation but also on formulations usable in these microsystems. Over the years we developed methods to understand how to optimize these formulations up to the creation of commercial products usable for a wide range of applications.
Creating life in the laboratory? A dialectical approach to bottom-up biology
Daphne Broeks1, Yogi Hendlin2, Hub Zwart2
1: Radboud University, The Netherlands; 2: Erasmus University Rotterdam, The Netherlands
A key impetus of modern science in general and of synthetic biology in particular has been to bridge the divide between the inorganic and the organic, between in vitro and in vivo, between the artificial and the natural: creating life in the laboratory, as it is phrased in the popular domain. Whereas the focus used to be on analysis (from living cell to genes, nucleotides, amino acids, etc.), the focus now shifts to synthesis. Bottom-up biology aims to understand biological processes via in vitro assembly of basic components in cell-like entities. Dialectical thinkers such as Hegel and Friedrich Engels already discerned a potential continuity (rather than an insurmountable discontinuity) between chemical processes and life and predicted that science would one day envision the technological reproducibility of life in vitro. Will this deepen our understanding of life, allowing us to learn from nature / mimic nature and develop sustainable technologies more compatible with living systems? Or will these artificial entities fail to capture the “aura” of life?
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