Abstract
Conventional reporter gene technology and histological methods cannot routinely be used to track the in vivo behavior of embryonic stem (ES) cells longitudinally after cellular transplantation. Here we describe a protocol for monitoring the in vivo survival, proliferation, and migration of ES cells without necessitating animal sacrifice. Stable ES cell lines containing double fusion (DF; enhanced green fluorescent protein and firefly luciferase) or triple fusion (TF; monomeric red fluorescent protein, firefly luciferase, and herpes simplex virus thymidine kinase) reporter genes can be established within 4–6 weeks by lentiviral transduction followed by fluorescence-activated cell sorting. The cell fate and behavior of these DF or TF ES cells can subsequently be tracked noninvasively by bioluminescence and microPET imaging for a prolonged period of time.
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Acknowledgments
This work was supported by a Bio-X graduate student fellowship (ASL), a Howard Hughes Medical Institute research fellowship (ASL), R21 HL091453 (JCW), and R21/R33 HL089027 (JCW).
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Lee, A.S., Wu, J.C. (2011). Imaging of Embryonic Stem Cell Migration In Vivo. In: Filippi, MD., Geiger, H. (eds) Stem Cell Migration. Methods in Molecular Biology, vol 750. Humana Press. https://doi.org/10.1007/978-1-61779-145-1_7
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DOI: https://doi.org/10.1007/978-1-61779-145-1_7
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