Intraspinal delivery of neurotrophin-3 using neural stem cells genetically modified by recombinant retrovirus.
Neural stem cells have been shown to participate in the repair of experimental CNS disorders. To examine their potential in spinal cord repair, we used retroviral vectors to genetically modify a clone of neural stem cells, C17, to overproduce neurotrophin-3 (NT-3). The cells were infected with a retrovirus construct containing the NT-3.IRES.lacZ/neo sequence and cloned by limiting dilution and selection for lacZ expression. We studied the characteristics of the modified neural stem cells in vitro and after transplantation into the intact spinal cord of immunosuppressed adult rats. Our results show that: (i) most of the genetically modified cells express both NT-3 and lacZ genes with a high coexpression ratio in vitro and after transplantation; and (ii) large numbers of the xenografted cells survive in the spinal cord of adult rats for at least 2 months, differentiate into neuronal and glial phenotypes, and migrate for long distances. We conclude that genetically modified neural stem cells, acting as a source of neurotrophic factors, have the potential to participate in spinal cord repair.
Liu
Y
Y
Department of Neurobiology and Anatomy, Medical College of Pennsylvania Hahnemann University, Philadelphia 19129, USA.
Himes
B T
BT
Solowska
J
J
Moul
J
J
Chow
S Y
SY
Park
K I
KI
Tessler
A
A
Murray
M
M
Snyder
E Y
EY
Fischer
I
I
D000818:Animals; D002454:Cell Differentiation; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D002999:Clone Cells / Q000502:physiology; D005260:Female; D015870:Gene Expression / Q000235:genetics; D005819:Genetic Markers; D007278:Injections, Spinal; D009414:Nerve Growth Factors / Q000096:biosynthesis*; D009474:Neurons / Q000378:metabolism / Q000637:transplantation*; D020933:Neurotrophin 3; D051381:Rats; D017207:Rats, Sprague-Dawley; D011995:Recombination, Genetic / Q000235:genetics; D012190:Retroviridae / Q000235:genetics; D013119:Spinal Cord Injuries / Q000628:therapy*; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000378:metabolism; D014162:Transfection / Q000235:genetics; D014183:Transplantation, Heterologous
D005819:Genetic Markers; D009414:Nerve Growth Factors; D020933:Neurotrophin 3