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

Author:

Pérez-Sosa Camilo12ORCID,Sanluis-Verdes Anahí1,Waisman Ariel2,Lombardi Antonella2ORCID,Rosero Gustavo1,Greca Alejandro La2,Bhansali Shekhar3,Bourguignon Natalia13,Luzzani Carlos2,Pérez Maximiliano. S.43,Miriuka Santiago2,Lerner Betiana13ORCID

Affiliation:

1. National Technological University (UTN), IREN Center, Buenos Aires, Argentina

2. National Scientific and Technical Research Council (CONICET) - Foundation for the Fight Against Childhood Neurological Diseases, (LIAN-CONICET-FLENI), FLENI Escobar Headquarters, Route 9 Km 53, 1625, Belén de Escobar, Buenos Aires, Argentina

3. Department of Electrical and Computer Engineering, Florida International University, Miami, FL 33174, USA

4. University of Buenos Aires (UBA), Institute of Biomedical Engineering, Paseo Colon 850, C1428EGA Buenos Aires, Argentina

Abstract

Microfluidic tools have recently made possible many advances in biological and biomedical research. Research in fields such as physics, engineering, chemistry and biology have combined to produce innovation in microfluidics which has positively impacted diverse areas such as nucleotide sequencing, functional genomics, single-cell studies, single molecules assays and biomedical diagnostics. Among these areas, regenerative medicine and stem cells have benefited from microfluidics since these tools have had a profound impact on their applications. In this study, we present a high-performance droplet-based system for transfecting individual human-induced pluripotent stem cells. We will demonstrate that this system has great efficiency in single cells and captured droplets, like other microfluidic methods but with lower cost. Moreover, 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 and cell therapy, among other potential applications.

Funder

CONICET

Publisher

The Royal Society

Subject

Multidisciplinary

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