Transgenic human ES and iPS reporter cell lines for identification and selection of pluripotent stem cells in vitro
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 cl...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2014-09-01
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| Series: | Stem Cell Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1873506114000658 |
| _version_ | 1811203794113921024 |
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| author | Dmitry A. Ovchinnikov Drew M. Titmarsh Patrick R.J. Fortuna Alejandro Hidalgo Samah Alharbi Deanne J. Whitworth Justin J. Cooper-White Ernst J. Wolvetang |
| author_facet | Dmitry A. Ovchinnikov Drew M. Titmarsh Patrick R.J. Fortuna Alejandro Hidalgo Samah Alharbi Deanne J. Whitworth Justin J. Cooper-White Ernst J. Wolvetang |
| author_sort | Dmitry A. Ovchinnikov |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-12T03:01:14Z |
| format | Article |
| id | doaj.art-d7ed9f79b774455283e162555ff5e793 |
| institution | Directory Open Access Journal |
| issn | 1873-5061 1876-7753 |
| language | English |
| last_indexed | 2024-04-12T03:01:14Z |
| publishDate | 2014-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Stem Cell Research |
| spelling | doaj.art-d7ed9f79b774455283e162555ff5e7932022-12-22T03:50:40ZengElsevierStem Cell Research1873-50611876-77532014-09-0113225126110.1016/j.scr.2014.05.006Transgenic human ES and iPS reporter cell lines for identification and selection of pluripotent stem cells in vitroDmitry A. Ovchinnikov0Drew M. Titmarsh1Patrick R.J. Fortuna2Alejandro Hidalgo3Samah Alharbi4Deanne J. Whitworth5Justin J. Cooper-White6Ernst J. Wolvetang7Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD 4072, AustraliaAustralian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD 4072, AustraliaAustralian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD 4072, AustraliaAustralian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD 4072, AustraliaAustralian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD 4072, AustraliaSchool of Veterinary Sciences, The University of Queensland, Gatton, QLD 4343, AustraliaAustralian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD 4072, AustraliaAustralian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD 4072, AustraliaOptimization 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.http://www.sciencedirect.com/science/article/pii/S1873506114000658 |
| spellingShingle | Dmitry A. Ovchinnikov Drew M. Titmarsh Patrick R.J. Fortuna Alejandro Hidalgo Samah Alharbi Deanne J. Whitworth Justin J. Cooper-White Ernst J. Wolvetang Transgenic human ES and iPS reporter cell lines for identification and selection of pluripotent stem cells in vitro Stem Cell Research |
| title | Transgenic human ES and iPS reporter cell lines for identification and selection of pluripotent stem cells in vitro |
| title_full | Transgenic human ES and iPS reporter cell lines for identification and selection of pluripotent stem cells in vitro |
| title_fullStr | Transgenic human ES and iPS reporter cell lines for identification and selection of pluripotent stem cells in vitro |
| title_full_unstemmed | Transgenic human ES and iPS reporter cell lines for identification and selection of pluripotent stem cells in vitro |
| title_short | Transgenic human ES and iPS reporter cell lines for identification and selection of pluripotent stem cells in vitro |
| title_sort | transgenic human es and ips reporter cell lines for identification and selection of pluripotent stem cells in vitro |
| url | http://www.sciencedirect.com/science/article/pii/S1873506114000658 |
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