A UTF1-based selection system for stable homogeneously pluripotent human embryonic stem cell cultures.
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.
Tan
Shen Mynn
SM
School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, 637551 and ES Cell International Pte Ltd, 11 Biopolis Way, #05-06 Helios, 138667 Singapore.
Wang
Siew Tein
ST
Hentze
Hannes
H
Dröge
Peter
P
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D005855:Germ Layers / Q000166:cytology; D006801:Humans; D008297:Male; D051379:Mice; D016513:Mice, SCID; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology*; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism; D019076:Transgenes
D015415:Biomarkers; D009687:Nuclear Proteins; D015534:Trans-Activators; C484738:UTF1 protein, human