Simple, efficient, and reproducible gene transfection of mouse embryonic stem cells by magnetofection

Chang Hyun Lee, Eun Young Kim, Kilsoo Jeon, Jin Cheol Tae, Keum Sil Lee, Yeon Ok Kim, Mi Young Jeong, Cheol Won Yun, Dong Kee Jeong, Somi K. Cho, Jae Hoon Kim, Hyo Yeon Lee, Key Zung Riu, Ssang Goo Cho, Se Pill Park

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

Embryonic stem (ES) cells are recognized as an excellent cell culture model for studying developmental mechanisms and their therapeutic modulations. The aim of this work was to define whether using magnetofection was an efficient way to manipulate stem cells genetically without adversely affecting their proliferation or self-renewal capacity. We compared our magnetofection results to those of a conservative method using FuGENE 6. Using enhanced green fluorescent protein (eGFP) as a reporter gene in D3 mouse ES (mES) cells, we found that magnetofection gave a significantly higher efficiency (45%) of gene delivery in stem cells than did the FuGENE 6 method (15%), whereas both demonstrated efficienct transfection in NIH-3T3 cells (60%). Although the transfected D3 (D3-eGFP) mES cells had undergone a large number of passages (>50), a high percentage of cells retained ES markers such as Oct-4 and stage-specific embryonic antigen-1 (SSEA-1). They also retained the ability to form embryoid bodies and differentiated in vitro into cells of the three germ layers. eGFP expression was sustained during stem cell proliferation and differentiation. This is the first transfection report using magnetofection in ES cells. On the basis of our results, we conclude that magnetofection is an efficient and reliable method for the introduction of foreign DNA into mouse ES cells and may become the method of choice.

Original languageEnglish
Pages (from-to)133-141
Number of pages9
JournalStem cells and development
Volume17
Issue number1
DOIs
Publication statusPublished - 2008 Feb 1

ASJC Scopus subject areas

  • Hematology
  • Developmental Biology
  • Cell Biology

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    Lee, C. H., Kim, E. Y., Jeon, K., Tae, J. C., Lee, K. S., Kim, Y. O., Jeong, M. Y., Yun, C. W., Jeong, D. K., Cho, S. K., Kim, J. H., Lee, H. Y., Riu, K. Z., Cho, S. G., & Park, S. P. (2008). Simple, efficient, and reproducible gene transfection of mouse embryonic stem cells by magnetofection. Stem cells and development, 17(1), 133-141. https://doi.org/10.1089/scd.2007.0064