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

41 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

Fingerprint

Stem cells
Transfection
Embryonic Stem Cells
Genes
Stem Cells
CD15 Antigens
Embryoid Bodies
Germ Layers
NIH 3T3 Cells
Reporter Genes
Cell Differentiation
Cell Culture Techniques
Cell Proliferation
Cell proliferation
Mouse Embryonic Stem Cells
DNA
Cell culture
enhanced green fluorescent protein
Modulation
Therapeutics

ASJC Scopus subject areas

  • Hematology

Cite this

Lee, C. H., Kim, E. Y., Jeon, K., Tae, J. C., Lee, K. S., Kim, Y. O., ... 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

Simple, efficient, and reproducible gene transfection of mouse embryonic stem cells by magnetofection. / Lee, Chang Hyun; Kim, Eun Young; Jeon, Kilsoo; Tae, Jin Cheol; Lee, Keum Sil; Kim, Yeon Ok; Jeong, Mi Young; Yun, Cheol-Won; Jeong, Dong Kee; Cho, Somi K.; Kim, Jae Hoon; Lee, Hyo Yeon; Riu, Key Zung; Cho, Ssang Goo; Park, Se Pill.

In: Stem Cells and Development, Vol. 17, No. 1, 01.02.2008, p. 133-141.

Research output: Contribution to journalArticle

Lee, CH, Kim, EY, Jeon, K, Tae, JC, Lee, KS, Kim, YO, Jeong, MY, Yun, C-W, Jeong, DK, Cho, SK, Kim, JH, Lee, HY, Riu, KZ, Cho, SG & Park, SP 2008, 'Simple, efficient, and reproducible gene transfection of mouse embryonic stem cells by magnetofection', Stem Cells and Development, vol. 17, no. 1, pp. 133-141. https://doi.org/10.1089/scd.2007.0064
Lee, Chang Hyun ; Kim, Eun Young ; Jeon, Kilsoo ; Tae, Jin Cheol ; Lee, Keum Sil ; Kim, Yeon Ok ; Jeong, Mi Young ; Yun, Cheol-Won ; Jeong, Dong Kee ; Cho, Somi K. ; Kim, Jae Hoon ; Lee, Hyo Yeon ; Riu, Key Zung ; Cho, Ssang Goo ; Park, Se Pill. / Simple, efficient, and reproducible gene transfection of mouse embryonic stem cells by magnetofection. In: Stem Cells and Development. 2008 ; Vol. 17, No. 1. pp. 133-141.
@article{323016a684064a8b858058b61a62d2d1,
title = "Simple, efficient, and reproducible gene transfection of mouse embryonic stem cells by magnetofection",
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.",
author = "Lee, {Chang Hyun} and Kim, {Eun Young} and Kilsoo Jeon and Tae, {Jin Cheol} and Lee, {Keum Sil} and Kim, {Yeon Ok} and Jeong, {Mi Young} and Cheol-Won Yun and Jeong, {Dong Kee} and Cho, {Somi K.} and Kim, {Jae Hoon} and Lee, {Hyo Yeon} and Riu, {Key Zung} and Cho, {Ssang Goo} and Park, {Se Pill}",
year = "2008",
month = "2",
day = "1",
doi = "10.1089/scd.2007.0064",
language = "English",
volume = "17",
pages = "133--141",
journal = "The BMJ",
issn = "0730-6512",
publisher = "Kluwer Academic Publishers",
number = "1",

}

TY - JOUR

T1 - Simple, efficient, and reproducible gene transfection of mouse embryonic stem cells by magnetofection

AU - Lee, Chang Hyun

AU - Kim, Eun Young

AU - Jeon, Kilsoo

AU - Tae, Jin Cheol

AU - Lee, Keum Sil

AU - Kim, Yeon Ok

AU - Jeong, Mi Young

AU - Yun, Cheol-Won

AU - Jeong, Dong Kee

AU - Cho, Somi K.

AU - Kim, Jae Hoon

AU - Lee, Hyo Yeon

AU - Riu, Key Zung

AU - Cho, Ssang Goo

AU - Park, Se Pill

PY - 2008/2/1

Y1 - 2008/2/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=39449092235&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=39449092235&partnerID=8YFLogxK

U2 - 10.1089/scd.2007.0064

DO - 10.1089/scd.2007.0064

M3 - Article

VL - 17

SP - 133

EP - 141

JO - The BMJ

JF - The BMJ

SN - 0730-6512

IS - 1

ER -