Details
| Originalsprache | Englisch |
|---|---|
| Seiten (von - bis) | 21637-21646 |
| Seitenumfang | 10 |
| Fachzeitschrift | ACS Omega |
| Jahrgang | 9 |
| Ausgabenummer | 19 |
| Frühes Online-Datum | 3 Mai 2024 |
| Publikationsstatus | Veröffentlicht - 14 Mai 2024 |
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).
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in: ACS Omega, Jahrgang 9, Nr. 19, 14.05.2024, S. 21637-21646.
Publikation: Beitrag in Fachzeitschrift › Artikel › Forschung › Peer-Review
}
TY - JOUR
T1 - Microfluidic Transfection System and Temperature Strongly Influence the Efficiency of Transient Transfection
AU - Dehne, Michaela
AU - Neidinger, Simon Valentin
AU - Stark, Michael
AU - Adamo, Antonia Camilla
AU - Kraus, Xenia
AU - Färber, Nicolas
AU - Westerhausen, Christoph
AU - Bahnemann, Janina
N1 - Publisher Copyright: © 2024 The Authors. Published by American Chemical Society
PY - 2024/5/14
Y1 - 2024/5/14
N2 - 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).
AB - 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).
UR - http://www.scopus.com/inward/record.url?scp=85192468016&partnerID=8YFLogxK
U2 - 10.1021/acsomega.4c02590
DO - 10.1021/acsomega.4c02590
M3 - Article
AN - SCOPUS:85192468016
VL - 9
SP - 21637
EP - 21646
JO - ACS Omega
JF - ACS Omega
IS - 19
ER -