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Exp Cell Res. 2015 Jan 1;330(1):178-85. doi: 10.1016/j.yexcr.2014.07.020. Epub 2014 Aug 4.

A simple and efficient method for transfecting mouse embryonic stem cells using polyethylenimine.

Author information

1
Department of Integrative Biology, University of Colorado Denver, S4111, 1201 Fifth Street, Denver, CO 80204, United States.
2
Department of Chemistry, University of Colorado Denver, 1201 Fifth Street, Denver, CO 80204, United States.
3
Department of Integrative Biology, University of Colorado Denver, S4111, 1201 Fifth Street, Denver, CO 80204, United States. Electronic address: christopher.phiel@ucdenver.edu.

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.

KEYWORDS:

Mouse embryonic stem cells; Polyethylenimine; Transfection

PMID:
25102378
PMCID:
PMC4268182
DOI:
10.1016/j.yexcr.2014.07.020
[Indexed for MEDLINE]
Free PMC Article
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