Mesencephalic human neural progenitor cells transplanted into the neonatal hemiparkinsonian rat striatum differentiate into neurons and improve motor behaviour

Abstract

Neural stem cell transplantation is a promising strategy for the treatment of neurodegenerative diseases. To evaluate the differentiation potential of human neural progenitor cells (hNPCs) as a prerequisite for clinical trials, we intracerebrally transplanted in vitro expanded fetal mesencephalic hNPCs into hemiparkinsonian rats. On postnatal day one (P1), 17 animals underwent a unilateral intraventricular 6-hydroxydopamine injection into the right lateral ventricle. At P3, animals ( n = 10) received about 100 000 hNPCs (1 µ L) in the right striatum. Five weeks after birth, animals underwent behaviour tests prior to fixation, followed by immunohistochemistry on brain slices for human nuclei, glial fibrillary acidic protein, S100 β , neuronal nuclei antigen, neuron-specific enolase and tyrosine hydroxylase. Compared with the apomorphine-induced rotations in the lesioned-only group (7.4 ± 0.5 min ) showed a significant therapeutic improvement.Additionally, in the cylinder test, the lesioned-only animals preferred to use the ipsilateral forepaw. Conversely, the lesioned and transplanted animals showed no significant side bias similar to untreated control animals.Transplanted human nuclei-immunoreactive cells were found to survive and migrate up to 2000 µ m into the host parenchyma, many containing the pan-neuronal markers neuronal nuclei antigen and neuron-specific enolase. In the striatum, tyrosine hydroxylase-immunoreactive somata were also found, indicating a dopaminergic differentiation capacity of transplanted hNPCs in vivo . However, the relative number of tyrosine hydroxylase-immunoreactive neurons in vivo seemed to be lower than in corresponding in vitro differentiation. To minimize donor tissue necessary for transplantation, further investigations will aim to enhance dopaminergic differentiation of transplanted cells in vivo .

Mesencephalic Human Neural Progenitor Cells Transplanted into the Adult Hemiparkinsonian Rat Striatum Lack Dopaminergic Differentiation but Improve Motor Behavior

“…Here, we transplanted hNPCs derived from the ventral mesencephalic area of an 8-week-old human embryo into a DAdepleted striatum of adult rats. These cells have previously been shown to differentiate into TH-ir neurons accompanied by a behavioral benefit when transplanted into the lesioned striatum of juvenile animals [Hovakimyan et al, 2006]. In contrast to juvenile recipients, in the present parallel study, hNPC differentiation into DAergic neurons was not found, supporting the view of the environment as an instructive cue for neuronal specification and differentiation.…”

“…At the same time point, the sham control group (10 rats) received an injection of vehicle alone (1 l DMEM) using the same coordinates as for the transplantation group. Cells from the same passage and cell preparation were transplanted both in juvenile [Hovakimyan et al, 2006] and adult animals. To obtain similar conditions with our previous parallel transplantation study using these cells [Hovakimyan et al, 2006], no immunosuppression was performed.…”

“…NPCs isolated from the ventral mesencephalon of an elective aborted human fetus at 8 weeks gestational age were expanded in culture as described elsewhere [Hovakimyan et al, 2006]. If not specified otherwise, all material used for cell culturing was obtained from Invitrogen.…”

Differential Proteomics of the Cerebral Cortex of Juvenile, Adult and Aged Rats: An Ontogenetic Study

“…Several studies [24][25][26] focus on the transplantation of immortalized progenitor cells in 6-OHDA Hemiparkinson animal models [24,25] and the differences in the abundance of this area of the brain at different points of development [26]. Hovakimyan et al [24] and Haas et al [25] as well as Schwarz and Freed [27] showed that the micro-environment of neonatal brains seems to be important for the differentiation of transplanted neuronal progenitors. Therefore, a further characterization of the abundance patterns of proteins during development in the Cx of the rat brain might provide a better understanding of neuronal development.…”

The Use of Stem Cells in the Treatment of Parkinsons Disease

“…NSCs that can be harvested from the embryo around embryonic day (E) 14 -E15 in rodents [42,43,44] or 13 weeks post-fertilization in humans [45,46], are multipotent cells that have the ability to regenerate and also to give rise to neurons, oligodendrocytes and astrocytes that represent the three major cell lineages of the CNS [47]. Undifferentiated human fetal NSCs transplanted into the SNc of a MPTP ((1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine)-induced primate model of PD were able to survive, integrate and subsequently reduce some behavioral deficits [46].…”