Comparative Study of Methods for Administering Neural Stem/Progenitor Cells to Treat Spinal Cord Injury in Mice

Abstract

To investigate potential cures for spinal cord injury (SCI), several researchers have transplanted neural stem/ progenitor cells (NS/PCs) into the injured spinal cord by different procedures, including intralesional (IL), intrathecal (IT), and intravenous (IV) injection. However, there are no reports quantifying or comparing the number of cells successfully transplanted to the lesion site by each procedure in vivo. The purpose of the present study was to determine the optimal method of cell transplantation to the SCI site in terms of grafted cell survival and safety. For this purpose, we developed mouse NS/PCs that expressed a novel Venusluciferase fusion protein that enabled us to detect a minimum of 1,000 grafted cells in vivo by bioluminescence imaging (BLI). After inducing contusive SCI at the T10 level in mice, NS/PCs were transplanted into the injured animals three different ways: by IL, IT, or IV injection. Six weeks after the transplantation, BLI analysis showed that in the IL group, the luminescence intensity of the grafted cells had decreased to about 10% of its initial level, and appeared at the site of injury. In the IT group, the luminescence of the grafted cells, which was distributed throughout the entire subarachnoid space immediately after transplantation, was detected at the injured site 1 week later, and by 6 weeks had gradually decreased to about 0.3% of its initial level. In the IV group, no grafted cells were detected at the site of injury, but all of these mice showed luminescence in the bilateral chest, suggesting pulmonary embolism. In addition, one third of these mice died immediately after the IV injection. In terms of grafted cell survival and safety, we conclude that the IL application of NS/PCs is the most effective and feasible method for transplanting NS/PCs into the SCI site. INTRODUCTIONstem/progenitor cell (NS/PC) transplantation is considered one of the most promising, because the transplantation of NS/PCs into rodent SCI models (13,15,27,42, Because the adult central nervous system (CNS) has little potential for regeneration, spinal cord injury (SCI) 44,48) and of human neural stem/progenitor cells (NS/ PCs) into a marmoset SCI model (28) have been shown usually results in severe damage, leading to paraplegia, tetraplegia, or worse. Several strategies have been used to promote functional recovery. From the viewpoint of clinical trials, it is critical to to develop new therapies that would allow patients to regain the use of their paralyzed limbs. Because cell determine how best to apply cells to the injured spinal cord. Three application methods for cell transplantation transplantation is one of the potential therapies, various kinds of cells have been used as the transplantation source for SCI are used: intralesional (IL), intrathecal (IT), and intravenous (IV). Some previous studies have sought to for SCI (14,18,(32)(33)(34)37,38,46 compare different cell transplantation methods for treatcentrated virus was 1 × 10 8 to 2 × 10 8 transducing units per milliliter (TU/ml) when...

Significance of Remyelination by Neural Stem/Progenitor Cells Transplanted into the Injured Spinal Cord

“…2A) and was detectable by the IVIS bioimaging system. We also confirmed the ffLuc transduction into wt-NS/PCs, as previously reported [32]. Next, shi-NS/PCs or wt-NS/PCs were transplanted into the lesion site of NOD/SCID mice 9 days after contusive SCI.…”

“…This vector enabled the detection of grafted cells as strong bioluminescent ffLuc signals in live SCI mice and as fluorescent Venus signals using anti-green fluorescent protein antibody in fixed spinal-cord sections. This virus was obtained as previously reported [32]. The concentrated virus was added to the culture medium to infect primary NS/PCs derived from shiverer or wild-type mice (multiplicity of injection ¼ 1.0).…”

“…The concentrated virus was added to the culture medium to infect primary NS/PCs derived from shiverer or wild-type mice (multiplicity of injection ¼ 1.0). The lentivirally transduced NS/PCs (5 Â 10 5 cells per two microliters) were transplanted into the lesion epicenter 9 days after SCI as previously reported [11,12,15,16,[31][32][33].…”

Intrathecal transplantation of stem cells by lumbar puncture for thoracic spinal cord injury in the rat

“…[2][3][4] The intrathecal technique of cell injection has also been used in other animal models. [5][6][7] However, the phenotype of LP transplanted neural stem/progenitor cells (NSPCs) and bone marrow-derived mesenchymal stromal cells (BMSCs) and their spatial distribution along the intact and injured spinal cord has not been reported or quantitated. LP is a minimally invasive method of cell delivery, and stem cells may be well suited for LP transplantation because of their responsiveness to signals from the injured CNS.…”

Neural Stem Cells Directly Differentiated from Partially Reprogrammed Fibroblasts Rapidly Acquire Gliogenic Competency

“…To evaluate the in vivo differentiation properties and safety of the diNSCs, we transplanted dissociated diNSC-derived neurospheres, which had been transduced by a lentivirus to express the Luciferase and Venus fluorescent proteins [52], into the striatum of C57/bl6j mice, as previously described [8,9]. BLI analysis [12], which detects luciferase photon signals only from living cells, showed that the cells were engrafted 28 days after their transplantation (Fig.…”