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Schmitt Oliver

S. Beckmann, S.J.P. Haas, S. Petrov, O. Schmitt, A. Wree

Institute of Anatomy, University of Rostock, Rostock, Germany


Cell replacement strategies for the treatment of Parkinson´s disease are promising, but still discussed controversially. Immortalized progenitor cells could be of further therapeutic interest, because they have several advantages (ethical, availability and handling) compared to primary cells or embryonic stem cells. As shown by Anton et al. (1994, Exp Neurol, 127: 207-218) as well as in own experiments CSM14.1-cells did not differentiate into dopaminergic neurons when grafted in the adult rat striatum. However, the immortalized progenitor cell line CSM14.1 has the capability to differentiate into a dopaminergic fate in vitro (Haas and Wree, 2002, J Anat, 201: 61-69). The objective of the presented study was to investigate the differentiation potential of these cells in the striatum of neonatal parkinsonian rats. On postnatal day one, 23 rats were bilaterally lesioned by a stereotactic intraventricular 6-OHDA-injection (2 x 1 µl, containing 12 µg 6-OHDA, coordinates according to bregma: AP –0.6; L ± 0.8; V (dura) –2.1). On P3, 6 lesioned animals received a unilateral graft consisting of 100.000 PKH26-labelled CSM14.1-cells into the right caudate-putamen complex (CPu, coordinates: AP +0.7; L –1.8; V –2.9). Five weeks after transplantation, forelimb preference was evaluated with the cylinder test in age matched intact controls (n = 10), bilaterally lesioned (n = 17) and unilaterally transplanted (after lesion) animals (n = 6). Six weeks after birth, animals were perfused and brain sections were processed for immunohistochemistry to detect changes in tyrosine hydroxylase (TH). Transplanted brains were treated as described earlier to investigate the differentiation of the PKH26-labelled transplanted cells by immunohistochemistry (Haas et al., 2000, Acta Histochem, 102: 273-280). No paw preference (right : left) was observed in intact (ratio 15 : 15 ± 1.76) and bilaterally lesioned (ratio 14.8 : 15.2 ± 2.1) animals, whereas lesioned animals, which received a unilateral cell graft in the right CPu showed a significant preference for the left paw (ratio 12.8 : 17.2 ± 1.72, p = 0.042, U-test). This kind of lesion leads to a nearly complete bilateral striatal dopaminergic deafferentiation as demonstrated by TH-immunohistochemistry. Transplants in animals showed a strong graft-derived TH-immunoreactivity in the right CPu with many TH-immunoreactive cell bodies. These grafted cells migrated approximately 1200 µm into the host parenchyma. Fluorescence and confocal laser scanning microscopy confirmed their differentiation into TH- and NeuN-immunoreactive neurons or astrocytes containing GFAP or S100 beta. Therefore, in all probability the preferential forelimb use could be due to a dopaminergic reinnervation by the grafted cells and further densitometric measurement and statistical analysis of TH-immunoreactivity corroborates this observation.

Keywords: Parkinson, Progenitor cells, Transplantation, 6-OHDA lesion, Rat.

Funding sources: Medical Faculty of the University of Rostock