Abstract:
:Optimization of pluripotent stem cell expansion and differentiation is facilitated by biological tools that permit non-invasive and dynamic monitoring of pluripotency, and the ability to select for an undifferentiated input cell population. Here we report on the generation and characterisation of clonal human embryonic stem (HES3, H9) and human induced pluripotent stem cell lines (UQEW01i-epifibC11) that have been stably modified with an artificial EOS(C3+) promoter driving expression of EGFP and puromycin resistance-conferring proteins. We show that EGFP expression faithfully reports on the pluripotency status of the cells in these lines and that antibiotic selection allows for an efficient elimination of differentiated cells from the cultures. We demonstrate that the extinction of the expression of the pluripotency reporter during differentiation closely correlates with the decrease in expression of conventional pluripotency markers, such as OCT4 (POU5F1), TRA-1-60 and SSEA4 when screening across conditions with various levels of pluripotency-maintaining or differentiation-inducing signals. We further illustrate the utility of these lines for real-time monitoring of pluripotency in embryoid bodies and microfluidic bioreactors.
journal_name
Stem Cell Resjournal_title
Stem cell researchauthors
Ovchinnikov DA,Titmarsh DM,Fortuna PR,Hidalgo A,Alharbi S,Whitworth DJ,Cooper-White JJ,Wolvetang EJdoi
10.1016/j.scr.2014.05.006subject
Has Abstractpub_date
2014-09-01 00:00:00pages
251-61issue
2eissn
1873-5061issn
1876-7753pii
S1873-5061(14)00065-8journal_volume
13pub_type
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journal_title:Stem cell research
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journal_title:Stem cell research
pub_type: 杂志文章
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