Transplantation of mouse embryonic stem cell-derived oligodendrocytes in the murine model of globoid cell leukodystrophy

Abstract

IntroductionGloboid cell leukodystrophy (GLD) is a severe disorder of the central and peripheral nervous system caused by the absence of galactocerebrosidase (GALC) activity. Cell-based therapies are highly promising strategies for GLD. In this study, G-Olig2 mouse embryonic stem cells (ESCs) were induced into oligodendrocyte progenitor cells (OPCs) and were implanted into the brains of twitcher mice, an animal model of GLD, to explore the therapeutic potential of the cells.MethodsThe G-Olig2 ESCs were induced into OPCs by using cytokines and a multi-step differentiation procedure. Oligodendrocyte markers were detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry. The toxicity of psychosine to OPCs was determined by a cell proliferation assay kit. The GALC level of OPCs was also examined. OPCs were labeled with Dir and transplanted into the brains of twitcher mice. The transplanted cells were detected by in-Vivo Multispectral Imaging System and real-time PCR. The physiological effects of twitcher mice were assessed.ResultsOligodendrocyte markers were expressed in OPCs, and 76% ± 5.76% of the OPCs were enhanced green fluorescent protein (eGFP)-positive, eGFP was driven by the Olig2 promoter. The effect of psychosine on cell viability indicated that OPCs were more resistant to psychosine toxicity. The GALC level of OPCs was 10.0 ± 1.23 nmol/hour per mg protein, which was significantly higher than other cells. Dir-labeled OPCs were injected into the forebrain of post-natal day 10 twitcher mice. The transplanted OPCs were myelin basic protein (MBP)-positive and remained along the injection tract as observed by fluorescent microscopy. The level of the Dir fluorescent signal and eGFP mRNA significantly decreased at days 10 and 20 after injection, as indicated by in-Vivo Multispectral Imaging System and real-time PCR. Because of poor cell survival and limited migration ability, there was no significant improvement in brain GALC activity, MBP level, life span, body weight, and behavioral deficits of twitcher mice.ConclusionsESC-derived OPC transplantation was not sufficient to reverse the clinical course of GLD in twitcher mice.

Leukodystrophies

“…A recent cell therapy study for mice with Krabbe disease failed, because accumulation of galactosyl-ceramide and its toxic derivative psychosine killed the maturing oligodendrocytes after transplantation. 21 Our studies in patients with vanishing white matter disease showed that hyaluronan is increased in the brain extracellular matrix, 22 which impedes maturation of oligodendrocyte progenitors and may also affect transplanted oligodendrocyte progenitors.…”

Treatment for Krabbe's disease: Finding the combination

“…Twenty days after injection, the level of myelin was measured, as was the distribution of implanted donor cells. There was no increase in the level of myelin basic protein in treated mice, and the donor cells remained localized to the injection site (Kuai el al., ). In addition, there was no improvement in behavioral assays measuring stride length or twitching severity, and life span of treated mice was not significantly different from that of untreated mice or those injected with saline (Kuai et al, ).…”

Osthole Enhances the Therapeutic Efficiency of Stem Cell Transplantation in Neuroendoscopy Caused Traumatic Brain Injury

“…The transplantation process we used has been previously described. [50][51][52] Because we imitated microcosmic surgery in the brain, we transplanted cells immediately after brain injury. To avoid additional injury, we injected cells into the same hole with a depth of 2 mm, to make sure transplantation had a minimal effect on the injury we created.…”