title: A simple and efficient method for transfecting mouse embryonic stem cells using polyethylenimine.
abstract: Mouse embryonic stem cells (ESCs) can be transfected by electroporation, liposomal reagents, and viral transduction methods. The cationic polymer polyethylenimine (PEI) has been shown to transfect a variety of differentiated mammalian cell types, including mouse ESCs, but existing methods require the use of additional equipment that is not readily accessible to most labs. Here we describe conditions that permit for the efficient transfection of mouse ESCs with low cytotoxicity and without the need for specialized equipment. Our goal was to devise a protocol for the PEI-mediated transfection of mouse ESCs that was comparable in ease to commercial transfection reagents. For these studies, we compared PEI transfection efficiency and cytotoxicity to a well-known liposomal transfection reagent, Lipofectamine2000(™) (LF2K), using fluorescence microscopy, flow cytometry, cell viability assays, and Western blotting. We provide evidence that PEI transfection of mouse ESCs compares favorably to LF2K. Our optimized protocol for efficient transfection of mouse ESCs with PEI is detailed in this report.
authors.affiliation: Department of Integrative Biology, University of Colorado Denver, S4111, 1201 Fifth Street, Denver, CO 80204, United States.
authors.forename: Colleen M
authors.initials: CM
authors.lastname: Bartman
authors.affiliation: Department of Integrative Biology, University of Colorado Denver, S4111, 1201 Fifth Street, Denver, CO 80204, United States.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Egelston
authors.affiliation: Department of Chemistry, University of Colorado Denver, 1201 Fifth Street, Denver, CO 80204, United States.
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Ren
authors.affiliation: Department of Integrative Biology, University of Colorado Denver, S4111, 1201 Fifth Street, Denver, CO 80204, United States.
authors.forename: Raibatak
authors.initials: R
authors.lastname: Das
authors.affiliation: Department of Integrative Biology, University of Colorado Denver, S4111, 1201 Fifth Street, Denver, CO 80204, United States. Electronic address: christopher.phiel@ucdenver.edu.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Phiel
mesh_terms: D000818:Animals; D053595:Embryonic Stem Cells / Q000187:drug effects / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D039904:Pluripotent Stem Cells / Q000187:drug effects / Q000378:metabolism; D011094:Polyethyleneimine / Q000494:pharmacology*; D014162:Transfection / Q000379:methods*
chemical_list: D011094:Polyethyleneimine
keywords: Mouse embryonic stem cells; Polyethylenimine; Transfection
references.citation: Development. 2009 Jul;136(14):2311-22
references.pmid: 19542351
references.citation: J Control Release. 2012 Mar 10;158(2):304-11
references.pmid: 22138069
references.citation: J Liposome Res. 2003 Feb;13(1):29-32
references.pmid: 12725725
references.citation: Biomaterials. 2010 Jan;31(1):124-32
references.pmid: 19818490
references.citation: Nanomedicine (Lond). 2014 May;9(7):999-1017
references.pmid: 24063366
references.citation: Anal Bioanal Chem. 2010 Aug;397(8):3173-8
references.pmid: 20549496
references.citation: Stem Cells. 2004;22(4):531-43
references.pmid: 15277699
references.citation: Transgenic Res. 2000 Jun;9(3):229-32
references.pmid: 11032372
references.citation: Biomaterials. 2011 Sep;32(25):5924-33
references.pmid: 21600648
references.citation: Adv Med Sci. 2014 Mar;59(1):7-12
references.pmid: 24797966
references.citation: Biologicals. 2007 Jun;35(3):165-71
references.pmid: 17084092
references.citation: Int J Nanomedicine. 2013;8:1935-46
references.pmid: 23737665
references.citation: Nanomedicine. 2013 Nov;9(8):1181-91
references.pmid: 23727125
references.citation: Stem Cells Dev. 2008 Feb;17(1):133-41
references.pmid: 18271700
references.citation: BMC Biotechnol. 2010 Feb 08;10:9
references.pmid: 20144189
references.citation: Mol Pharm. 2012 Sep 4;9(9):2698-709
references.pmid: 22862421
references.citation: Int J Biochem Mol Biol. 2011;2(4):318-27
references.pmid: 22187666
references.citation: Int J Nanomedicine. 2012;7:4637-48
references.pmid: 22942645
references.citation: Nucleic Acids Res. 2002 Jan 15;30(2):E9
references.pmid: 11788735
references.citation: Biotechnol Bioeng. 2008 Feb 15;99(3):721-7
references.pmid: 17680657
references.citation: Mol Pharm. 2009 Sep-Oct;6(5):1333-42
references.pmid: 19552481
references.citation: Biomaterials. 2011 Sep;32(26):6174-82
references.pmid: 21696819
references.citation: Cell Tissue Res. 2012 Mar;347(3):689-99
references.pmid: 22277991
references.citation: Biomacromolecules. 2012 May 14;13(5):1429-37
references.pmid: 22455481
references.citation: Proc Natl Acad Sci U S A. 1995 Aug 1;92(16):7297-301
references.pmid: 7638184
references.citation: Biotechnol Prog. 2005 Jan-Feb;21(1):148-53
references.pmid: 15903252
references.citation: Exp Cell Res. 2002 Sep 10;279(1):34-9
references.pmid: 12213211
references.citation: Biomacromolecules. 2008 Mar;9(3):818-27
references.pmid: 18247565
references.citation: Cytotechnology. 2010 Jan;62(1):73-82
references.pmid: 20309632
references.citation: Acta Biomater. 2012 Nov;8(11):3921-31
references.pmid: 22820309
references.citation: Bull Exp Biol Med. 2011 May;151(1):126-9
references.pmid: 22442817
references.citation: Biomaterials. 2012 Sep;33(27):6485-94
references.pmid: 22687755
references.citation: J Cell Mol Med. 2011 Sep;15(9):1989-98
references.pmid: 20629995
references.citation: Mamm Genome. 2012 Oct;23(9-10):600-10
references.pmid: 22961258
references.citation: J Biol Chem. 2010 Mar 12;285(11):7957-63
references.pmid: 20080974
references.citation: Gene Ther. 2002 Jan;9(2):102-9
references.pmid: 11857068
references.citation: Biol Chem. 2003 May;384(5):737-47
references.pmid: 12817470