OBJECTIVETo explore the feasibility for therapy of spinal cord injury (SCI) by genetic engineering neural stem cell (NSC) modified by lentiviral vector.

METHODSFollowing the construction of the genetic engineering NSC modified by lentivirus to secrete both neurotrophic factor-3 (NT-3) and green fluorescence protein (GFP), hemisection of spinal cord at the level of T10 was performed in 56 adult Wistar rats that were randomly divided into 4 groups (n = 14), namely 3 therapeutic groups and 1 control group. The therapeutic groups were dealed with NSC, genetic engineering NSC, and concentrated lentiviral supernatant which carries both GFP and NT-3, respectively. Then used fluorescence microscope to detect the transgenic expression in vitro and in vivo, migration of the grafted cells in vivo, and used the Basso, Beattie, and Bresnahan (BBB) open-field locomotor test to assess the recovery of function.

RESULTSThe transplanted cells could survive for long time in vivo and migrate for long distance. The stable transgenic expression could be detected in vivo. The hindlimb function of the injured rats in 3 therapeutic groups, especially those dealed with genetic engineering NSC, improved obviously.

CONCLUSIONIt is feasible to combine NSC with lentivirus for the repair of SCI. NSC modified by lentivirus to deliver NT-3, acting as a source of neurotrophic factors and function cell in vivo, has the potential to participate in spinal cord repair.

Animals ; Cell Line ; Embryonic Stem Cells ; transplantation ; Female ; Genetic Engineering ; methods ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Lentivirus ; genetics ; Neurons ; transplantation ; Neurotrophin 3 ; genetics ; Rats ; Rats, Wistar ; Spinal Cord Injuries ; therapy

OBJECTIVETo explore the feasibility to construct genetic engineering human neural stem cells (hNSCs) mediated by lentivirus to express multigene in order to provide a graft source for further studies of spinal cord injury (SCI).

METHODSHuman neural stem cells from the brain cortex of human abortus were isolated and cultured, then gene was modified by lentivirus to express both green fluorescence protein (GFP) and rat neurotrophin-3 (NT-3); the transgenic expression was detected by the methods of fluorescence microscope, dorsal root ganglion of fetal rats and slot blot.

RESULTSGenetic engineering hNSCs were successfully constructed. All of the genetic engineering hNSCs which expressed bright green fluorescence were observed under the fluorescence microscope. The conditioned medium of transgenic hNSCs could induce neurite flourishing outgrowth from dorsal root ganglion (DRG). The genetic engineering hNSCs expressed high level NT-3 which could be detected by using slot blot.

CONCLUSIONSGenetic engineering hNSCs mediated by lentivirus can be constructed to express multigene successfully.

Animals ; Cell Differentiation ; Cells, Cultured ; Feasibility Studies ; Gene Expression ; Genetic Engineering ; methods ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; Humans ; Immunohistochemistry ; Lentivirus ; genetics ; Microscopy, Fluorescence ; Neurons ; metabolism ; Rats ; Stem Cell Transplantation ; Stem Cells ; metabolism ; Transgenes