Non-Invasive Cell Tracking with Brighter and Red-Transferred Luciferase for Potential Application in Stem Cell Therapy

Abstract

This study investigated the safety of a novel cell-labeling technology with mKATE and Renilla reniformis luciferase (mKATE-renLUC) and assessed the efficacy on tracking implanted human placental stromal cells (PSC) in an erectile dysfunction (ED) animal model. Human PSC were labeled with mKATE-renLUC using a lentivirus. Cell viability, apoptosis, proliferation, migration, surface marker expression and differentiation potential of the labeled PSC were evaluated and compared with non-labeled PSC. The paracrine profile of labeled cells was examined using an angiogenesis protein array. The brightness and duration of labeled cells with different densities were evaluated. An ED rat model was established and labeled PSC were injected into cavernosal tissue of the penis. The migration and distribution of transplanted PSC were monitored using an IVIS imaging system in real time. Implanted PSC were identified in isolated tissues via detection of mKATE fluorescence. The cell viability, morphology, proliferation, migration, surface marker expression and differentiation potential of mKATE-renLUC-labeled PSC were similar to those of non-labeled cells in vitro (no statistical difference p>0.05). Similar expressions of trophic factors were found between labeled and non-labeled PSC. The migration and distribution of PSC expressing renLUC were tracked in vivo using IVIS imaging system. mKATE-positive PSC were detected in penile, kidney, prostate and hepatic tissues using histological methods. This labeling technology provides a safe and effective cell-tracking approach with a brighter fluorophore and codon-optimized luciferase.

Human placental-derived stem cell therapy ameliorates experimental necrotizing enterocolitis

“…hPSC localization was first examined in an exploratory pilot cohort of 5 NEC-hPSC pups using dual-labeled hPSC for in vivo cell tracking and immunohistochemical analysis. As previously shown (36), hPSC were dual-tagged via lentiviral infection with a luciferase from Renilla reniformis (renLUC, for in vivo cell tracking) and mKATE (for IHC). NEC-hPSC pups received dual-labeled hPSC (referred to as mKATE hPSC) for the first cell injection (at 32 h) followed by untagged hPSC in the second cell injection.…”

“…NEC-hPSC pups received dual-labeled hPSC (referred to as mKATE hPSC) for the first cell injection (at 32 h) followed by untagged hPSC in the second cell injection. For in vivo imaging on day 4, the luciferase substrate (Colenterazine 200 mg/kg) was intraperitoneally injected and in vivo imaging system (IVIS) images were obtained 5 min after substrate injection (36). An untreated NEC pup was used as an imaging control.…”

“…For immunohistochemistry, the intestinal tissue was processed as described in Immunostaining and Quantifications and immunostained with a rabbit polyclonal anti-tRFP antibody that recognized mKATE (Cat No. AB233, Evrogen, NY) (36).…”

Prospects of Therapeutic Target and Directions for Ischemic Stroke

“…Bioluminescence imaging visualizes photons emitted by luciferase enzyme reactions in live animals. To enable bioluminescence imaging, however, stem cells must be genetically modified to express the luciferase enzyme before transplantation [ 147 , 148 ] Stem cells can be labeled with nanoparticles such as superparamagnetic iron oxide nanoparticles, quantum dots, and pebbles in order to chase transplanted stem cells using MRI [ 149 , 150 , 151 ]. One study suggested the use of NSCs for the treatment of neurological diseases in a nonhuman primate stroke animal model.…”

Coelenterazine-Dependent Luciferases as a Powerful Analytical Tool for Research and Biomedical Applications

“…However, the main application of RLuc is luciferase genetic reporter assay that has become an invaluable and routine tool for molecular biology research, including identification and characterization of protein functional variants [ 147 , 148 ], investigations of gene expression [ 148 , 149 , 150 ], transcription factors [ 151 , 152 , 153 , 154 , 155 ], receptor activity [ 156 ], miRNA expression [ 157 , 158 , 159 ], monitoring mRNA splicing events in cells [ 160 , 161 ], virus investigations [ 162 , 163 , 164 ], drug discovery [ 165 , 166 ], and identification and evaluation of virus inhibitors [ 167 , 168 ]. Moreover, RLuc is widely used for bioluminescence imaging of different processes in vivo [ 169 , 170 , 171 ] and guiding drug delivery to cells [ 172 , 173 ].…”