Multimodality Molecular Imaging of Cardiac Cell Transplantation: Part I. Reporter Gene Design, Characterization, and Optical in Vivo Imaging of Bone Marrow Stromal Cells after Myocardial Infarction

Abstract

4 The complete list of authors is at the end of this article.q RSNA, 2016 Purpose:To use multimodality reporter-gene imaging to assess the serial survival of marrow stromal cells (MSC) after therapy for myocardial infarction (MI) and to determine if the requisite preclinical imaging end point was met prior to a follow-up large-animal MSC imaging study. Materials and Methods:Animal studies were approved by the Institutional Administrative Panel on Laboratory Animal Care. Mice (n = 19) that had experienced MI were injected with bone marrow-derived MSC that expressed a multimodality triple fusion (TF) reporter gene. The TF reporter gene (fluc2-egfp-sr39ttk) consisted of a human promoter, ubiquitin, driving firefly luciferase 2 (fluc2), enhanced green fluorescent protein (egfp), and the sr39tk positron emission tomography reporter gene. Serial bioluminescence imaging of MSC-TF and ex vivo luciferase assays were performed. Correlations were analyzed with the Pearson product-moment correlation, and serial imaging results were analyzed with a mixed-effects regression model.

Mesenchyme modulates three-dimensional, collective cell migration of liver cellsin vitro- a role for TGFß pathway

“…HepG2 cells (p10) cultivated at 200,000 cells/well in a 6-well plate, were transfected with a third generation lentivirus bearing a transgene (ubiquitin-fluc2-egfp (Human Ubiquitin promoter, Firefly luciferase 2, enhanced green fluorescent protein)), as done previously, (Parashurama, Ahn et al 2016) at an MOI of 5.…”

Engineering molecular imaging strategies for regenerative medicine

“…For in vivo imaging with BLI, a substrate is required that has to be stable, bioavailable, have favorable pharmacokinetics. Fluc has been used for a wide range of cell imaging applications, including neural stem cell tracking in the brain, cardiac cell transplantation, assessment of gene delivery, epigenetic modulation of reporter expression, immunosuppression efficacy, graft versus host disease, evaluation of tissue scaffolds, whole body HSC reconstitution liver cell therapy, encapsulated in vivo cell viability, decellularized liver matrix, oxidative stress within transplanted cells, differentiation, and multimodality imaging . This information is summarized in Figure and Table .…”

“…Dosing with pharmaceutical can be done based on mass of the patient (or the amount of receptor available), but because molecular targets of cell therapies are complex, it is unclear how to perform exact dosing. In a recent set of publications, we demonstrated experimentally an idea of how molecular imaging in small animals and large animals could be connected, to improve the clinical translation of cell therapies . In this case, multimodality RG, bearing eGFP (enhanced GFP), Fluc2, and hsv1 mutant (sr39tk), for fluorescence, bioluminescence, and PET RG imaging, were transduced into MSC.…”

“…Fluc has been used for a wide range of cell imaging applications, including neural stem cell tracking in the brain, 158 cardiac cell transplantation, 165,166 assessment of gene delivery, 167 epigenetic modulation of reporter expression, 168 immunosuppression efficacy, 169 graft versus host disease, 170 evaluation of tissue scaffolds, 171 whole body HSC reconstitution 172 liver cell therapy, 173 encapsulated in vivo cell viability, 174 decellularized liver matrix, 175 oxidative stress within transplanted cells, 176 differentiation, [177][178][179] and multimodality imaging. 180 This information is summarized in Figure 5 and Table 2 immediate early promoter enhancer with chicken beta-actin/rabbit beta globin hybrid promoter, or the (CAG) promoter, which drives a fluc-egfp fusion protein. 172 Because all the cells in the mouse express RG strongly, these transgenic mice have been actively used as donor mice in cell transplantation experiments.…”

“…In a recent set of publications, we demonstrated experimentally an idea of how molecular imaging in small animals and large animals could be connected, to improve the clinical translation of cell therapies. 82,180 In this case, multimodality RG, bearing eGFP (enhanced GFP), Fluc2, and hsv1 mutant (sr39tk), for fluorescence, bioluminescence, and PET RG imaging, were transduced into MSC. These MSCs were tested in small animal disease models and Fluc signal was associated with cell…”