title: A High-Throughput Microfluidic Platform for Mammalian Cell Transfection and Culturing.
abstract: Mammalian synthetic biology could be augmented through the development of high-throughput microfluidic systems that integrate cellular transfection, culturing, and imaging. We created a microfluidic chip that cultures cells and implements 280 independent transfections at up to 99% efficiency. The chip can perform co-transfections, in which the number of cells expressing each protein and the average protein expression level can be precisely tuned as a function of input DNA concentration and synthetic gene circuits can be optimized on chip. We co-transfected four plasmids to test a histidine kinase signaling pathway and mapped the dose dependence of this network on the level of one of its constituents. The chip is readily integrated with high-content imaging, enabling the evaluation of cellular behavior and protein expression dynamics over time. These features make the transfection chip applicable to high-throughput mammalian protein and synthetic biology studies.
authors.affiliation: Institute of Bioengineering, School of Engineering, École Polytechnique Fédérale de, Lausanne, Switzerland.
authors.forename: Kristina
authors.initials: K
authors.lastname: Woodruff
authors.affiliation: Institute of Bioengineering, School of Engineering, École Polytechnique Fédérale de, Lausanne, Switzerland.
authors.forename: Sebastian J
authors.initials: SJ
authors.lastname: Maerkl
mesh_terms: D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D000071677:Histidine Kinase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D056656:Lab-On-A-Chip Devices; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D061848:Optical Imaging / Q000295:instrumentation / Q000379:methods*; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism*; D015398:Signal Transduction; D014162:Transfection
chemical_list: D000071677:Histidine Kinase