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Gene & Protein in Disease Transfection methods for TK6 cells

the interaction between cationic lipids and DNA which Carolina A and T State University for their overwhelming
facilitates the delivery of DNA into the cells [63,64] . Our study support. Furthermore, we thanked Dr. Perpetua Muganda
found that transfecting TK6 cells with Lipofectamine LTX for her continued support and supervision.
yielded a relatively high percent transfection efficiency,
cell viability, and luciferase activity at 24 and 36 h post- Funding
transfections, suggesting that lipofectamine could be This work was supported in part by a National Institute of
used in our system. High luciferase activity at 36 h post- Environmental Health Sciences AREA grant ES019306,
transfection of lipofectamine LTX has also been observed and National Institute of General Medical Sciences MBRS
in primary human umbilical vein endothelial and mice SCORE grant GM076530.
cells [59,65] .
The Amaxa Nucleofector 96-Well Shuttle System is Conflict of interest
a fully automated high throughput system to transfect There are no conflicts of interest to declare.
difficult-to-transfect cell lines and primary cells in the
96-well format. The system is an attractive primary Author contributions
experimental tool due to its simplicity and reproducible Conceptualization: Akamu Jude Ewunkem
results. Amaxa Nucleofector Shuttle has been shown to Formal analysis: Akamu Jude Ewunkem
deliver successfully high transfection efficiency in several Investigation: Akamu Jude Ewunkem
cell lines [66-69] . However, Amaxa Nucleofector Shuttle Methodology: Akamu Jude Ewunkem
has shown some very poor transfection results in other Writing – original draft: Akamu Jude Ewunkem
cells, suggesting that cell type has a major influence Writing – review & editing: Akamu Jude Ewunkem, Agee
on transfection efficiency of cells transfected with Kyle.
Nucleofector [70-73] . In this study, transfecting TK6 with 0.4
µg of plasmid DNA using reagent SF and program DS 137 Ethics approval and consent to participate
was associated with high transfection efficiency (~80%)
and luciferase activity (RLU = 5.1 × 10 ) with acceptable Not applicable.
5
cell toxicity (~15%). Chicaybam et al. observed similar Consent for publication
results when T-lymphocytes were nucleofected at the same
transfection conditions . However, nucleofection was Not applicable.
[74]
more toxic to our cells compared to the other transfection
methods used due to long-lasting pulses or polarization of Availability of data
the cells from the electric field [75,76] . Amaxa nucleofection All data generated or analyzed during this study are
was identified as the optimal transfection reagent for included in this published article.
transfecting TK6 cells due to its higher luciferase activity,
high transfection efficiency, and acceptable cell viability References
percentage. 1. Zu Y, Huang S, Lu Y, et al., 2016, Size specific transfection
to mammalian cells by micropillar array electroporation. Sci
5. Conclusion Rep, 6(1): 38661–38610.
Our results show that of the three tested reagents, Amaxa 2. Alabdullah AA, Al-Abdulaziz B, Alsalem H, et al., 2019,
96-well Nucleofection Shuttle System using Solution SF Estimating transfection efficiency in differentiated and
delivered the best transfection results in traditionally hard- undifferentiated neural cells. BMC Res Notes, 12(1): 225.
to-transfect cell lines such as TK6 human lymphoblasts. https://doi.org/10.1186/s13104-019-4249-5
Transfection with Amaxa Nucleofection Shuttle System
using solution SF yields high luciferase activity and 3. Lieberman JR, Ghivizzani SC, Evans CH, 2002, Gene
transfection efficiency and is accompanied by acceptable transfer approaches to the healing of bone and cartilage. Mol
Ther, 6(2): 141–147.
cell toxicity.
https://doi.org/10.1006/mthe.2000.0663
Acknowledgments 4. Avci-Adali M, Behring A, Keller T, et al., 2014, Optimized
The author would like special thanks to the faculty and staff conditions for successful transfection of human endothelial
in Biological Sciences Department, Winston Salem State cells with in vitro synthesized and modified mRNA for
University, Department of Biology North Carolina A and induction of protein expression. J Biol Eng, 8(1): 8.
T State University, Department of Applied Science, North https://doi.org/10.1186/1754-1611-8-8.

Volume 2 Issue 2 (2023) 9 https://doi.org/10.36922/gpd.0353