Table 3.
Preclinical studies utilizing reporter gene tracking of stem cell therapies.
| Cell Therapy | Purpose of Imaging | Reporter Gene (RG) Expressed | Imaging Modality Used | Method of RG transfer | Refs. |
|---|---|---|---|---|---|
| Adult Stem Cells/Tissue Resident Stem Cells | |||||
| Human and mouse HSCs | engraftment monitoring | human deoxycytidine kinase with three amino acid substitutions within the active site (hdCK3mut) | PET/CT | lentivirus | 200 |
| Immortalized human neural stem cell line (HB1.F3) | study of epigenetic silencing mechanisms of reporter genes in neural stem cells | hNIS | scintographic imaging | plasmid transfection | 201 |
| Immortalized human bone marrow-derived MSC line | monitoring of MSC homing to tumors and evaluation of their therapeutic potential as a transgenic reporter-expressing cell-based therapy | hNIS | scintographic imaging | plasmid transfection | 202 |
| Human MSCs | monitoring of MSC homing and therapeutic potential in breast cancer | hNIS | SPECT | adenovirus | 203 |
| Human MSCs | tracking of long-term fate and trafficking of MSCs | triple fusion protein: fLuc-mRFP-HSV1-sr39tk | BLI and PET/CT | lentivirus | 204 |
| Human MSCs | understanding MSC fate in tissue repair | mutant of dopamine type 2 receptor (D2R80A) | PET | lentivirus | 205 |
| Human MSCs | evaluating myocardial tracking potential with a PET reporter in small (rat) and large animal studies (swine) | HSV1-sr39tk | PET | adenovirus | 206 |
| hESCs and Their Differentiated Progeny | |||||
| Transplanted labeled hESCs/H9 line | tracking immune rejection | fusion protein: fLuc and EGFP | BLI | lentivirus | 207 |
| Human neural stem cells derived from hESCs/H7 line | tracking fate and function of grafted cells in a preclinical stroke model | triple fusion protein: mRFP-fLuc-HSV1-sr39tk | MRI and PET | lentivirus | 208 |
| hESCs | teratoma monitoring after transplant into chick embryos and mice (kidney capsule and muscle of peritoneum) | fLuc | BLI | plasmid transfection | 209 |
| Human ESCs/H9 line | determining application of genome editing for long-term molecular imaging of engrafted stem cells | polycistronic: EGFP/fLuc/hSSTR2 and polycistronic EGFP/fLuc/hNIS | BLI and PET | ZFN targeted at the AAVS1 locus | 210 |
| hESCs/H9 line and one patient derived hiPSC line and hESC-derived ECs and CMs | preclinical monitoring of teratomas and hESC-derived cardiac cells for cardiovascular research/regenerative medicine | triple fusion protein: fLuc-mRFP-HSV1-tk | BLI | ZFN targeted at the AAVS1 locus | 211 |
| hESC-derived CD34+ cells/H9 line | tracking engraftment/developmental of hESC-derived HSCs in vivo | fLuc | BLI | transfection of DNA transposon system | 212 |
| hESCs/H9 line | safety study: analysis number of contaminating undifferentiated hESCs required to yield a teratoma | fusion protein: fLuc-EGFP | BLI | lentivirus | 198 |
| hESC-derived MSCs | studied the distribution of human MSCs in a rat hindlimb ischemic injury model immediately after transplantation and also analyzed the recipient tissue response to transplanted cells | fLuc | BLI | lentivirus | 213 |
| hESCs and hESC-derived ECs/H9 line | comparison of MR and bioluminescence modalities for tracking of transplanted cell engraftment and longitudinal monitoring of cell fate | fusion protein: fLuc-EGFP | BLI | lentivirus | 214 |
| hESC-derived neural precursors/H9 line | monitoring of long-term viability and proliferation of hESC-derived neural precursor grafts in the brains of immunodeficient and immunocompetent mice | TGL fusion protein: HSV1-tk-EGFP-fLuc | BLI | lentivirus | 215 |
| hESC-derived skeletal myoblasts/H1 and H9 lines | assessment of long-term myoblast engraftment and survival with monitoring for teratoma formation | TGL fusion protein: HSV1-tk-EGFP-fLuc | BLI | lentivirus | 216 |
| hESCs/H1 and H9 | monitoring stem cell engraftment and teratoma formation | bicistronic fLuc and GFP and fusion of HSV1-tk-GFP | BLI and SPECT/CT | lentivirus | 199 |
| hiPSCs and Their Differentiated Progeny | |||||
| hiPSCs differentiated to motor neurons, HLCs, and macrophages | generation of reporter expressing hiPSCs suitable for differentiation into macrophages to track anti-fibrotic potential in vivo | ZsGreen | in vivo imaging not performeda | ZFN targeted at AAVS1 locus | 217 |
| hiPSC-derived HLCs | potential for tracking transplanted HLC populations in vivo | hNIS-EGFP fusion | SPECT/CT | lentivirus | 172 |
| hiPSC-derived neural stem/progenitor cells | determining the feasibility of tumor ablation following hiPSC-neural stem/progenitor cell (NS/PC) spinal cord transplantation utilizing immunoregulation | fusion protein Venus-fLuc | BLI | lentivirus | 218 |
| hiPSC-derived endothelial cells | analysis of potential of iPSC-derived ECs to promote perfusion of ischemic tissue in model of peripheral arterial disease | fusion protein: fLuc-EGFP | BLI | lentivirus | 219 |
| hiPSCs | evaluation of transplanted hiPSC survival, engraftment, and distribution of in a pig model of myocardial infarction | bicistronic rat NIS and Venus | SPECT/CT | plasmid transfection | 220 |
| hiPSCs | evaluating systems to purge residual hiPSCs before graft without compromising hematopoietic repopulation capability | fLuc | BLI | lentivirus | 221 |
| hiPSC-derived cardiomyocytes | assessment of relationship between transplanted cell number and engraftment rate in myocardial injury | bicistronic fLuc and GFP | BLI | lentivirus | 222 |
The table illustrates the range of preclinical stem cell therapy studies that have incorporated reporter gene-afforded in vivo imaging. Studies are classified based on type of stem cell, with details on the modality and purpose of in vivo tracking used as well as the reporter gene and method of construct integration. HSC, hematopoietic stem cell; hESC, human embryonic stem cell; hiPSC, human induced pluripotent stem cell; RG, reporter gene; EC, endothelial cell; CM, cardiomyocyte; HLC, hepatocyte-like cell; ZFN, zinc finger nuclease; fLuc, firefly luciferase; mRFP, monomeric red fluorescence protein; HSV1-tk, herpes simplex virus type 1 thymidine kinase; HSV1-sr39tk, truncated HSV1-sr39 thymidine kinase; hNIS, human sodium iodide symporter; hSSTR2, human somatostatin receptor 2; BLI, bioluminescence imaging.
a
Cited as a tool with the potential for macrophage in vivo tracking in the future.