@article {Love2011, author = {Zachary Love and Fangjing Wang and James Dennis and Amad Awadallah and Nicolas Salem and Yuan Lin and Andrew Weisenberger and Stan Majewski and Stanton Gerson and Zhenghong Lee}, title = {Imaging of Mesenchymal Stem Cell Transplant by Bioluminescence and PET}, volume = {48}, number = {12}, pages = {2011--2020}, year = {2007}, doi = {10.2967/jnumed.107.043166}, publisher = {Society of Nuclear Medicine}, abstract = {Dynamic measurements of infused stem cells generally require animal euthanasia for single-time-point determinations of engraftment. In this study, we used a triple-fusion reporter system for multimodal imaging to monitor human mesenchymal stem cell (hMSC) transplants. Methods: hMSCs were transduced with a triple-fusion reporter, fluc-mrfp-ttk (encoding firefly luciferase, monomeric red fluorescent protein, and truncated herpes simplex virus type 1 sr39 thymidine kinase) by use of a lentiviral vector. Transduced cells were assayed in vitro for the expression of each functional component of the triple-fusion reporter. Transduced and control hMSCs were compared for their potential to differentiate into bone, cartilage, and fat. hMSCs expressing the reporter were then loaded into porous, fibronectin-coated ceramic cubes and subcutaneously implanted into NOD-SCID mice along with cubes that were loaded with wild-type hMSCs and empty cubes. Mice were imaged repeatedly over 3 mo by bioluminescence imaging (BLI), and selected animals underwent CT and PET imaging. Results: Osteogenic, adipogenic, and chondrogenic potential assays revealed retained differentiation potentials between transduced and wild-type hMSCs. Signals from the cubes loaded with reporter-transduced hMSCs were visible by BLI over 3 mo. There was no signal from the empty or wild-type hMSC{\textendash}loaded control cubes. PET data provided confirmation of the quantitative estimation of the number of cells at one spot (cube). Cubes were removed from some animals, and histologic evaluations showed bone formation in cubes loaded with either reporter-transduced or wild-type hMSCs, whereas empty controls were negative for bone formation. Conclusion: The triple-fusion reporter approach resulted in a reliable method of labeling stem cells for investigation in small-animal models by use of both BLI and small-animal PET imaging. It has the potential for translation into future human studies with clinical PET.}, issn = {0161-5505}, URL = {https://jnm.snmjournals.org/content/48/12/2011}, eprint = {https://jnm.snmjournals.org/content/48/12/2011.full.pdf}, journal = {Journal of Nuclear Medicine} }