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Continuous monitoring of cell transfection efficiency with micropatterned substrates
Author(s) -
AzuajeHualde Enrique,
Rosique Melania,
CalatayudSanchez Alba,
BenitoLopez Fernando,
M. de Pancorbo Marian,
BasabeDesmonts Lourdes
Publication year - 2021
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.27783
Subject(s) - transfection , cell , flow cytometry , microbiology and biotechnology , stromal cell , mesenchymal stem cell , cell culture , biology , chemistry , biochemistry , genetics , cancer research
Abstract The effect of cell–cell contact on gene transfection is mainly unknown. Usually, transfection is carried out in batch cell cultures without control over cellular interactions, and efficiency analysis relies on complex and expensive protocols commonly involving flow cytometry as the final analytical step. Novel platforms and cell patterning are being studied to control cellular interactions and improve quantification methods. In this study, we report the use of surface patterning of fibronectin for the generation of two types of mesenchymal stromal cell patterns: single‐cell patterns without cell‐to‐cell contact, and small cell‐colony patterns. Both scenarios allowed the integration of the full transfection process and the continuous monitoring of thousands of individualized events by fluorescence microscopy. Our results showed that cell‐to‐cell contact clearly affected the transfection, as single cells presented a maximum transfection peak 6 h earlier and had a 10% higher transfection efficiency than cells with cell‐to‐cell contact.