Microfluidic Transfection System and Temperature Strongly Influence the Efficiency of Transient Transfection

verfasst von
Michaela Dehne, Simon Valentin Neidinger, Michael Stark, Antonia Camilla Adamo, Xenia Kraus, Nicolas Färber, Christoph Westerhausen, Janina Bahnemann
Abstract

For the process of transient transfection (TTF), DNA is often transported into the cells using polyplexes. The polyplex uptake and the subsequent transient expression of the gene of interest are of great importance for a successful transfection. In this study, we investigated a 3D-printed microfluidic system designed to facilitate direct TTF for suspension of CHO-K1 cells. The results demonstrate that this system achieves significantly better results than the manual approach. Furthermore, the effect of both post-transfection incubation time (t) and temperature (T) on polyplex uptake was explored in light of the membrane phase transitions. Attention was paid to obtaining the highest possible transfection efficiency (TFE), viability (V), and viable cell concentration (VCC). Our results show that transfection output measured as product of VCC and TFE is optimal for t = 1 h at T = 22 °C. Moreover, post-transfection incubation at T = 22 °C with short periods of increased T at T = 40 °C were observed to further increase the output. Finally, we found that around T = 19 °C, the TFE increases strongly. This is the membrane phase transition T of CHO-K1 cells, and those results therefore suggest a correlation between membrane order and permeability (and in turn, TFE).

Organisationseinheit(en)
Institut für Technische Chemie
Externe Organisation(en)
Universität Augsburg
Typ
Artikel
Journal
ACS Omega
Band
9
Seiten
21637-21646
Anzahl der Seiten
10
Publikationsdatum
14.05.2024
Publikationsstatus
Veröffentlicht
Peer-reviewed
Ja
ASJC Scopus Sachgebiete
Allgemeine Chemie, Allgemeine chemische Verfahrenstechnik
Elektronische Version(en)
https://doi.org/10.1021/acsomega.4c02590 (Zugang: Offen)