In vivo bioluminescent imaging of virus-mediated gene transfer and transduced cell transplantation in the intervertebral disc.
METHODS
Work presented here used a small animal model to demonstrate the feasibility and usefulness of in vivo bioluminescent imaging to studying degenerative disc disease.


OBJECTIVE
To determine the utility of in vivo bioluminescent imaging to monitor the temporal and spatial expression of genetically modified cells within the intervertebral disc of a rodent model.


BACKGROUND
Noninvasive imaging of genetically engineered cells in the spine has the advantage of allowing events to be tracked without killing the animal and can be used to follow the time course of a particular therapy. Results are presented on the use of Sprague-Dawley rats in experimental studies in which the luciferase reporter gene was delivered to the lumbar intervertebral disc through adenovirus-mediated or cell-based transfer techniques to demonstrate the feasibility to monitor gene expression noninvasively over time.


METHODS
Tissue culture, disc surgery, and in vivo bioluminescent imaging were used. The intervertebral disc of the rat was either injected in situ with an adenovirus containing the luciferase reporter gene or implanted with fat, bone marrow or intervertebral disc cells transduced ex vivo and contained in a bioresorbable carrier. Results were assessed with in vivo bioluminescent imaging at several time points.


CONCLUSIONS
Data from 11 animals were obtained with imaging up to 14 days. To our knowledge, this is the first description of in vivo bioluminescence imaging to study spinal conditions. We have characterized the relative expression of three cell types transduced with the Ad-luc virus by ex vivo transfection followed by cell implantation in the rat spine and compared them to one another and to direct infection of Ad-luc adenovirus in situ. Our results demonstrate the feasibility of tracing genetically altered cells in the spine. This technique has the potential to be used to noninvasively track the fate and expression of therapeutic genes within the spine of small animals used in disc research.
Leo
Brian M
BM
Department of Orthopaedic Surgery, University of Virginia School of Medicine, Charlottesville, VA, USA.
Li
Xudong
X
Balian
Gary
G
Anderson
D Greg
DG
D000256:Adenoviridae / Q000235:genetics*; D000273:Adipose Tissue / Q000166:cytology / Q000378:metabolism / Q000637:transplantation; D000464:Alginates; D000818:Animals; D016026:Bone Marrow Transplantation; D002478:Cells, Cultured; D004333:Drug Administration Routes; D005240:Feasibility Studies; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D007403:Intervertebral Disc / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation*; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D008163:Luminescent Measurements; D008297:Male; D023421:Models, Animal; D051381:Rats; D017207:Rats, Sprague-Dawley; D014161:Transduction, Genetic / Q000379:methods
D000464:Alginates; D006603:Hexuronic Acids; D020723:Glucuronic Acid; D008156:Luciferases