Noninvasive reporter gene imaging of human Oct4 (pluripotency) dynamics during the differentiation of embryonic stem cells in living subjects.
Purpose: Human pluripotency gene networks (PGNs), controlled in part by Oct4, are central to understanding pluripotent stem cells, but current fluorescent reporter genes (RGs) preclude noninvasive assessment of Oct4 dynamics in living subjects.Procedures: To assess Oc4 activity noninvasively, we eng...
| Publicado en: | Molecular Imaging & Biology Vol. 16; no. 6; pp. 865 - 877 |
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| Autores principales: | , , , , , , , |
| Formato: | research Journal Article |
| Publicado: |
Springer Nature
Dec2014
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=109764624&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 109764624 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 15361632 KJU jtl: Molecular Imaging & Biology issn: 15361632 maglogo: N pubinfo: dt: Dec2014 vid: 16 iid: 6 pid: 237 pub: Springer Nature place: New York, New York artinfo: ui: 109764624 NLM24845530 2012799696 10.1007/s11307-014-0744-1 NLM24845530 109764624 ppf: 865 ppct: 12 formats: fmt: @attributes: type: P tig: atl: Noninvasive reporter gene imaging of human Oct4 (pluripotency) dynamics during the differentiation of embryonic stem cells in living subjects. aug: au: Ahn, Byeong-Cheol Parashurama, Natesh Patel, Manish Ziv, Keren Bhaumik, Srabani Yaghoubi, Shahriar Shah Paulmurugan, Ramasamy Gambhir, Sanjiv Sam sug: subj: Molecular Imaging Methods Proteins Metabolism Stem Cells Physiology Cell Differentiation Physiology Cell Line Genes Microscopy Models, Biological Oxidoreductases Oxidoreductases Metabolism Proteins Rats Stem Cells ab: Purpose: Human pluripotency gene networks (PGNs), controlled in part by Oct4, are central to understanding pluripotent stem cells, but current fluorescent reporter genes (RGs) preclude noninvasive assessment of Oct4 dynamics in living subjects.Procedures: To assess Oc4 activity noninvasively, we engineered a mouse embryonic stem cell line which encoded both a pOct4-hrluc (humanized renilla luciferase) reporter and a pUbi-hfluc2-gfp (humanized firefly luciferase 2 fused to green fluorescent protein) reporter.Results: In cell culture, pOct4-hRLUC activity demonstrated a peak at 48 h (day 2) and significant downregulation by 72 h (day 3) (p=0.0001). Studies in living subjects demonstrated significant downregulation in pOct4-hRLUC activity between 12 and 144 h (p = 0.001) and between 12 and 168 h (p = 0.0003). pOct4-hRLUC signal dynamics after implantation was complex, characterized by transient upregulation after initial downregulation in all experiments (n = 10, p = 0.01). As expected, cell culture differentiation of the engineered mouse embryonic stem cell line demonstrated activation of mesendodermal, mesodermal, endodermal, and ectodermal master regulators of differentiation, indicating potency to form all three germ layers.Conclusions: We conclude that the Oct4-hrluc RG system enables noninvasive Oct4 imaging in cell culture and in living subjects. pubtype: Academic Journal doctype: research Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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