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Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/25490

Title: Molecular Imaging of Human Embryonic Stem Cells Stably Expressing Human PET Reporter Genes After Zinc Finger Nuclease-Mediated Genome Editing
Authors: Wolfs, Esther
Holvoet, Bryan
Ordovas, Laura
Breuls, Natacha
Helsen, Nicky
Schonberger, Matthias
Raitano, Susanna
Struys, Tom
Vanbilloen, Bert
Casteels, Cindy
Sampaolesi, Maurilio
Van Laere, Koen
Lambrichts, Ivo
Verfaillie, Catherine M.
Deroose, Christophe M.
Issue Date: 2017
Citation: JOURNAL OF NUCLEAR MEDICINE, 58(10), p. 1659-1665
Abstract: Molecular imaging is indispensable for determining the fate and persistence of engrafted stem cells. Standard strategies for transgene induction involve the use of viral vectors prone to silencing and insertional mutagenesis or the use of nonhuman genes. Methods: We used zinc finger nucleases to induce stable expression of human imaging reporter genes into the safe-harbor locus adeno-associated virus integration site 1 in human embryonic stem cells. Plasmids were generated carrying reporter genes for fluorescence, bioluminescence imaging, and human PET reporter genes. Results: In vitro assays confirmed their functionality, and embryonic stem cells retained differentiation capacity. Teratoma formation assays were performed, and tumors were imaged over time with PET and bioluminescence imaging. Conclusion: This study demonstrates the application of genome editing for targeted integration of human imaging reporter genes in human embryonic stem cells for longterm molecular imaging.
Notes: [Wolfs, Esther; Holvoet, Bryan; Breuls, Natacha; Vanbilloen, Bert; Casteels, Cindy; Van Laere, Koen; Deroose, Christophe M.] Katholieke Univ Leuven, Dept Imaging & Pathol, Mol Small Anim Imaging Ctr, Nucl Med & Mol Imaging, Leuven, Belgium. [Ordovas, Laura; Helsen, Nicky; Raitano, Susanna; Verfaillie, Catherine M.] Katholieke Univ Leuven, Stem Cell Inst, Leuven, Belgium. [Ordovas, Laura; Helsen, Nicky; Raitano, Susanna; Verfaillie, Catherine M.] Katholieke Univ Leuven, Stem Cell Biol & Embryol, Dept Dev & Regenerat, Leuven, Belgium. [Breuls, Natacha; Sampaolesi, Maurilio] Katholieke Univ Leuven, Translat Cardiomyol Lab, Dept Dev & Regenerat, Leuven, Belgium. [Schonberger, Matthias] Katholieke Univ Leuven, Dept Pharmaceut & Pharmacol Sci, Leuven, Belgium. [Struys, Tom; Lambrichts, Ivo] Univ Hasselt, Biomed Res Inst, Morphol Res Grp, Histol Lab, Diepenbeek, Belgium.
URI: http://hdl.handle.net/1942/25490
DOI: 10.2967/jnumed.117.189779
ISI #: 000411985900025
ISSN: 0161-5505
Category: A1
Type: Journal Contribution
Validation: ecoom, 2018
Appears in Collections: Research publications

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