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

Publikation: Beitrag in FachzeitschriftArtikelForschungPeer-Review

Autoren

  • Michaela Dehne
  • Simon Valentin Neidinger
  • Michael Stark
  • Antonia Camilla Adamo
  • Xenia Kraus
  • Nicolas Färber
  • Christoph Westerhausen
  • Janina Bahnemann

Organisationseinheiten

Externe Organisationen

  • Universität Augsburg
Forschungs-netzwerk anzeigen

Details

OriginalspracheEnglisch
Seiten (von - bis)21637-21646
Seitenumfang10
FachzeitschriftACS Omega
Jahrgang9
Ausgabenummer19
Frühes Online-Datum3 Mai 2024
PublikationsstatusVeröffentlicht - 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).

ASJC Scopus Sachgebiete

Zitieren

Microfluidic Transfection System and Temperature Strongly Influence the Efficiency of Transient Transfection. / Dehne, Michaela; Neidinger, Simon Valentin; Stark, Michael et al.
in: ACS Omega, Jahrgang 9, Nr. 19, 14.05.2024, S. 21637-21646.

Publikation: Beitrag in FachzeitschriftArtikelForschungPeer-Review

Dehne, M, Neidinger, SV, Stark, M, Adamo, AC, Kraus, X, Färber, N, Westerhausen, C & Bahnemann, J 2024, 'Microfluidic Transfection System and Temperature Strongly Influence the Efficiency of Transient Transfection', ACS Omega, Jg. 9, Nr. 19, S. 21637-21646. https://doi.org/10.1021/acsomega.4c02590
Dehne, M., Neidinger, S. V., Stark, M., Adamo, A. C., Kraus, X., Färber, N., Westerhausen, C., & Bahnemann, J. (2024). Microfluidic Transfection System and Temperature Strongly Influence the Efficiency of Transient Transfection. ACS Omega, 9(19), 21637-21646. https://doi.org/10.1021/acsomega.4c02590
Dehne M, Neidinger SV, Stark M, Adamo AC, Kraus X, Färber N et al. Microfluidic Transfection System and Temperature Strongly Influence the Efficiency of Transient Transfection. ACS Omega. 2024 Mai 14;9(19):21637-21646. Epub 2024 Mai 3. doi: 10.1021/acsomega.4c02590
Dehne, Michaela ; Neidinger, Simon Valentin ; Stark, Michael et al. / Microfluidic Transfection System and Temperature Strongly Influence the Efficiency of Transient Transfection. in: ACS Omega. 2024 ; Jahrgang 9, Nr. 19. S. 21637-21646.
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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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AU - Stark, Michael

AU - Adamo, Antonia Camilla

AU - Kraus, Xenia

AU - Färber, Nicolas

AU - Westerhausen, Christoph

AU - Bahnemann, Janina

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