Optical bioluminescence imaging of human ES cell progeny in the rodent CNS.
Pre-clinical efforts of grafting human embryonic stem cell (hESC)-derived neural precursors have been hampered by problems ranging from graft rejection to overgrowth and tumor formation. The ability to detect such potential complications sensitively and reliably in clinically relevant contexts will rest upon the implementation of suitable non-invasive imaging technologies for continuously probing graft survival, proliferation, and migration. Neural precursors were transduced ex vivo using a lentiviral-mediated gene delivery system expressing firefly D-luciferase, under the control of a cytomegalovirus promoter. Transduced cells revealed no loss of cellular morphology, proliferative capacity, or neural phenotype in vitro. As a novel approach to monitoring the fate of human grafts within the living brain, we adapted optical bioluminescence imaging to assess long-term graft viability in immunodeficient mouse models transplanted with genetically engineered human neural precursor cells. We additionally applied this technology to immunocompetent models for detecting and characterizing the time course of graft rejection. Using this strategy, we define statistically relevant imaging criteria that can predict graft rejection or overgrowth. In conclusion, our data suggest that optical bioluminescence imaging can serve as an essential tool for the development of hESC-based grafting strategies in the CNS.
Bradbury
Michelle S
MS
Department of RadiologyDevelopmental Biology ProgramNeurosurgery Service, Sloan Kettering Institute for Cancer Research, New York City, New York, USA.
Panagiotakos
Georgia
G
Department of RadiologyDevelopmental Biology ProgramNeurosurgery Service, Sloan Kettering Institute for Cancer Research, New York City, New York, USA.
Chan
Bill K
BK
Department of RadiologyDevelopmental Biology ProgramNeurosurgery Service, Sloan Kettering Institute for Cancer Research, New York City, New York, USA.
Tomishima
Mark
M
Department of RadiologyDevelopmental Biology ProgramNeurosurgery Service, Sloan Kettering Institute for Cancer Research, New York City, New York, USA.
Zanzonico
Pat
P
Department of RadiologyDevelopmental Biology ProgramNeurosurgery Service, Sloan Kettering Institute for Cancer Research, New York City, New York, USA.
Vider
Jelena
J
Department of RadiologyDevelopmental Biology ProgramNeurosurgery Service, Sloan Kettering Institute for Cancer Research, New York City, New York, USA.
Ponomarev
Vladimir
V
Department of RadiologyDevelopmental Biology ProgramNeurosurgery Service, Sloan Kettering Institute for Cancer Research, New York City, New York, USA.
Studer
Lorenz
L
Department of RadiologyDevelopmental Biology ProgramNeurosurgery Service, Sloan Kettering Institute for Cancer Research, New York City, New York, USA.
Tabar
Viviane
V
Department of RadiologyDevelopmental Biology ProgramNeurosurgery Service, Sloan Kettering Institute for Cancer Research, New York City, New York, USA.
D000818:Animals; D002478:Cells, Cultured; D003342:Corpus Striatum / Q000166:cytology*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005456:Fluorescent Dyes; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D006084:Graft Rejection / Q000503:physiopathology / Q000517:prevention & control; D006085:Graft Survival / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D007121:Immunocompetence / Q000276:immunology; D008156:Luciferases / Q000235:genetics; D008163:Luminescent Measurements / Q000379:methods*; D051379:Mice; D016513:Mice, SCID; D008856:Microscopy, Fluorescence / Q000379:methods; D055095:Optics and Photonics; D033581:Stem Cell Transplantation / Q000379:methods*; D014183:Transplantation, Heterologous
D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D008156:Luciferases