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Nucleic Acids Research Advance Access originally published online on September 12, 2007
Nucleic Acids Research 2007 35(18):e118; doi:10.1093/nar/gkm704
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Nucleic Acids Research, 2007, Vol. 35, No. 18 e118
© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Methods Online

A UTF1-based selection system for stable homogeneously pluripotent human embryonic stem cell cultures

Shen Mynn Tan1, Siew Tein Wang2, Hannes Hentze2 and Peter Dröge1,*

1School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, 637551 and 2ES Cell International Pte Ltd, 11 Biopolis Way, #05-06 Helios, 138667 Singapore

*To whom correspondence should be addressed. Tel: +65 6316 2809; Fax: +65 6791 3856; Email: pdroge{at}ntu.edu.sg or pdroege2001{at}yahoo.de

Received March 2, 2007. Revised August 7, 2007. Accepted August 24, 2007.

Undifferentiated transcription factor 1 (UTF1) was identified first in mouse embryonic stem cells and is also expressed in human embryonic and adult stem cells. UTF1 transcription ceases at the onset of differentiation, which clearly distinguishes it from less sensitive pluripotency markers, such as Oct4 or Nanog. We present here two transgenic hESC lines, named ZUN. Each line harbors one copy of the UTF1 promoter/enhancer driving a resistance gene and yielded highly homogeneous cultures under selection pressure, with a larger proportion of Oct4 and Sox2 positive cells. While ZUN cultures, like parental HUES8 cultures, retained the capacity to differentiate into tissues of all three germ layers using a SICD mouse teratoma model, they surprisingly exhibited an increased refractoriness to various differentiation cues in vitro. Together with its small size of only 2.4 kb for the entire cassette, these features render our selection system a powerful novel tool for many stem cell applications and human somatic cell reprogramming strategies.


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