An optimized protocol for the electroporation of NCI H929 multiple myeloma cells

Öz

Multiple myeloma cell lines are difficult to transfect with non-viral nucleic acid delivery methods. While electroporation is the most efficient tool for the transfection of most hard-to-transfect cells, human multiple myeloma cells differ in their permissiveness and each cell type require different electroporation conditions for an efficient transgene delivery. In this study, various parameters for NCI H929 human multiple myeloma cells are tested to generate an optimized electroporation protocol. Findings from this paper showed that besides the voltage and capacitance settings, cell count, the cell cycle status of cells, the amount of nucleic acid and removal of death cells all impacted the electroporation efficiency and viable cell count. These results are expected to serve as a starting point and a guide for researchers.

Anahtar Kelimeler

• electroporation
• multiple myeloma
• NCI H929
• transfection

Kaynakça

1. Batista Napotnik, T., Polajžer, T., & Miklavčič, D. (2021). Cell death due to electroporation - A review. Bioelectrochemistry (Amsterdam, Netherlands), 141, 107871. https://doi.org/10.1016/j.bioelechem.2021.107871
2. Biorad. Gene Pulser Xcell Electroporation System Instruction Manual (Issue 62). https://www.bio-rad.com/sites/default/files/2022-01/10000148530.pdf
3. Brito, J. L., Brown, N., & Morgan, G. J. (2010). Transfection of siRNAs in multiple myeloma cell lines. Methods Mol Biol, 623, 299–309. https://doi.org/10.1007/978-1-60761-588-0_19
4. Brunner, S., Sauer, T., Carotta, S., Cotten, M., Saltik, M., & Wagner, E. (2000). Cell cycle dependence of gene transfer by lipoplex, polyplex and recombinant adenovirus. Gene Therapy, 7(5), 401–407. https://doi.org/10.1038/sj.gt.3301102
5. Canoy, R. J., Andre, F., Shmakova, A., Wiels, J., Lipinski, M., Vassetzky, Y., & Germini, D. (2020). Easy and robust electrotransfection protocol for efficient ectopic gene expression and genome editing in human B cells. Gene Ther. https://doi.org/10.1038/s41434-020-00194-x
6. Chong, Z. X., Yeap, S. K., & Ho, W. Y. (2021). Transfection types, methods and strategies: a technical review. PeerJ, 9, e11165–e11165. https://doi.org/10.7717/peerj.11165 Fus-Kujawa, A., Prus, P., Bajdak-Rusinek, K., Teper, P., Gawron, K., Kowalczuk, A., & Sieron, A. L. (2021). An Overview of Methods and Tools for Transfection of Eukaryotic Cells in vitro. Frontiers in Bioengineering and Biotechnology, 9. https://www.frontiersin.org/article/10.3389/fbioe.2021.701031
7. Hattori, H., Tagawa, S., & Kitani, T. (1995). [Establishment and usage of human multiple myeloma cell line]. Nihon Rinsho, 53(3), 603–609. https://www.ncbi.nlm.nih.gov/pubmed/7699892
8. Hsu, C. Y., & Uludag, H. (2012). A simple and rapid nonviral approach to efficiently transfect primary tissue-derived cells using polyethylenimine. Nat Protoc, 7(5), 935–945. https://doi.org/10.1038/nprot.2012.038

9. Mizrahy, S., Hazan-Halevy, I., Dammes, N., Landesman-Milo, D., & Peer, D. (2017). Current Progress in Non-viral RNAi-Based Delivery Strategies to Lymphocytes. Molecular Therapy : The Journal of the American Society of Gene Therapy, 25(7), 1491–1500. https://doi.org/10.1016/j.ymthe.2017.03.001
10. Oyeleye, O., Ogundeji, S., Ola, S., & Omitogun, O. (2016). Basics of animal cell culture: Foundation for modern science. Biotechnology and Molecular Biology Reviews, 11, 6–16. https://doi.org/10.5897/BMBR2016.0261
11. Rols, M.-P. (2017). Parameters Affecting Cell Viability Following Electroporation In Vitro BT - Handbook of Electroporation (D. Miklavčič (ed.); pp. 1449–1465). Springer International Publishing. https://doi.org/10.1007/978-3-319-32886-7_149
12. Rueden, C. T., Schindelin, J., Hiner, M. C., DeZonia, B. E., Walter, A. E., Arena, E. T., & Eliceiri, K. W. (2017). ImageJ2: ImageJ for the next generation of scientific image data. BMC Bioinformatics, 18(1), 529. https://doi.org/10.1186/s12859-017-1934-z
13. Shih, T., De, S., & Barnes, B. J. (2019). RNAi Transfection Optimized in Primary Naïve B Cells for the Targeted Analysis of Human Plasma Cell Differentiation. Frontiers in Immunology, 10. https://www.frontiersin.org/article/10.3389/fimmu.2019.01652
14. Steinbrunn, T., Chatterjee, M., Bargou, R. C., & Stühmer, T. (2014). Efficient transient transfection of human multiple myeloma cells by electroporation - An appraisal. PLoS ONE, 9(6). https://doi.org/10.1371/journal.pone.0097443
15. Sun, Y., Pan, J., Zhang, N., Wei, W., Yu, S., & Ai, L. (2017). Knockdown of long non-coding RNA H19 inhibits multiple myeloma cell growth via NF-κB pathway. Scientific Reports, 7(1). https://doi.org/10.1038/s41598-017-18056-9