Loading…

Continuous monitoring of cell transfection efficiency with micropatterned substrates

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...

Full description

Saved in:
Bibliographic Details
Published in:Biotechnology and bioengineering 2021-07, Vol.118 (7), p.2626-2636
Main Authors: Azuaje‐Hualde, Enrique, Rosique, Melania, Calatayud‐Sanchez, Alba, Benito‐Lopez, Fernando, M. de Pancorbo, Marian, Basabe‐Desmonts, Lourdes
Format: Article
Language:English
Subjects:
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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. The effect of cell‐cell contact on gene transfection is mostly unknown. Herein, patterning of adherent cells was used to evaluate this effect on transfection efficiency. Two types of cell patterns were produced: single cells arrays, without cell‐cell contact, and small cell‐clusters arrays, where cells were in contact with each other. This approach simplified current methodologies, allowed to address the effect of cell‐cell contact, and also allowed the continuous monitoring of the transfected gene expression for more than 30 hours.
ISSN:0006-3592
1097-0290
DOI:10.1002/bit.27783