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

authored by: 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).
Organisation(s): Institute of Technical Chemistry
External Organisation(s): University of Augsburg
Type: Article
Journal: ACS Omega
Volume: 9
Pages: 21637-21646
No. of pages: 10
Publication date: 14.05.2024
Publication status: Published
Peer reviewed: Yes
ASJC Scopus subject areas: General Chemistry, General Chemical Engineering
Electronic version(s): https://doi.org/10.1021/acsomega.4c02590 (Access: Open)

BibTeX

@article{f06c353ff8cb43ec94225e96c7403cce,
title = "Microfluidic Transfection System and Temperature Strongly Influence the Efficiency of Transient Transfection",
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).",
author = "Michaela Dehne and Neidinger, {Simon Valentin} and Michael Stark and Adamo, {Antonia Camilla} and Xenia Kraus and Nicolas F{\"a}rber and Christoph Westerhausen and Janina Bahnemann",
note = "Publisher Copyright: {\textcopyright} 2024 The Authors. Published by American Chemical Society",
year = "2024",
month = may,
day = "14",
doi = "10.1021/acsomega.4c02590",
language = "English",
volume = "9",
pages = "21637--21646",
journal = "ACS Omega",
issn = "2470-1343",
publisher = "American Chemical Society",
number = "19",
}