Single cell transfection of human induced pluripotent stem cells using a droplet-based microfluidic system

Abstract

Microfluidic tools have recently made possible many advances in biological and biomedical research. Research fields such as Physics, Engineering, Chemistry and Biology have combined to produce innovation in Microfluidics which has positively impacted on areas as diverse as nucleotide sequence, functional genomics, single-cell studies, single molecules assays, and biomedical diagnostics. Among these areas regenerative medicine and stem cells have benefited from Microfluidics due to these tools have had a profound impact on their applications. In the study, we present a high-performance droplet-based system for transfecting individual humaninduced pluripotent stem cells. We show that this system has great efficiency in single cells and captured droplets, similar to other microfluidic methods and lower cost. We demonstrate that this microfluidic approach can be associated with the PiggyBac transposase-based system to increase its transfection efficiency. Our results provide a starting point for subsequent applications in more complex transfection systems, single-cell differentiation interactions, cell subpopulations, cell therapy, among other potential applications.

Microfluidic-Based Droplets for Advanced Regenerative Medicine: Current Challenges and Future Trends

“…Recently, new methods with a combination of microfluidics-derived droplets and conventional gene transfection techniques have emerged, with the ability to enhance the accuracy of transportation of exogenous genetic factors at a single-cell level [ 138 ]. For instance, using these methods, Zhan et al delivered an enhanced green fluorescent protein (EGFP) plasmid vector into Chinese hamster ovary (CHO) cells; they encapsulated CHO and EGFP plasmids inside droplets, which then passed through a pair of microelectrodes with a constant voltage.…”