Abstract
The treatment of Parkinson's disease by transplantation of dopaminergic (DA) neurons from human embryonic mesencephalic tissue is a promising approach. However, the origin of these cells causes major problems: availability and standardization of the graft. Therefore, the generation of unlimited numbers of DA neurons from various types of stem or progenitor cells has been brought into focus. A source for DA neurons might be conditionally immortalized progenitor cells. The temperature-sensitive immortalized cell line CSM14.1 derived from the mesencephalon of an embryonic rat has been used successfully for transplantation experiments. This cell line was analyzed by unbiased stereology of cell type specific marker proteins and 2D-gel electrophoresis followed by mass spectrometry to characterize the differentially expressed proteome. Undifferentiated CSM14.1 cells only expressed the stem cell marker nestin, whereas differentiated cells expressed GFAP or NeuN and tyrosine hydroxylase. An increase of the latter cells during differentiation could be shown. By using proteomics an explanation on the protein level was found for the observed changes in cell morphology during differentiation, when CSM14.1 cells possessed the morphology of multipolar neurons. The results obtained in this study confirm the suitability of CSM14.1 cells as an in vitro model for the study of neuronal and dopaminergic differentiation in rats.
Highlights
The motoric cardinal symptoms in Parkinson’s disease (PD) are caused by the degeneration of dopaminergic (DA) neurons
Immortalized CSM14.1 cells [5] were cultivated and expanded as described by Haas and Wree [9] in DMEM supplemented with 10% fetal calf serum (FCS), 100 Units mL−1 penicillin, and 100 μg mL−1 s3t3r∘eCp)t.omCyelclinpainssaagheuwmaisdidfioendeinecvuebryatothri(r9d5%daya.irT, o5%inCduOc2e, differentiation the amount of FCS was reduced to 1% and the temperature was risen to 39∘C—nonpermissive temperature [12, 13]
The type VI intermediate filament (IF) protein nestin is a widely-used marker for neuronal progenitor cells
Summary
The motoric cardinal symptoms (rigor, tremor, akinesia, and postural instability) in Parkinson’s disease (PD) are caused by the degeneration of dopaminergic (DA) neurons. Most of these dopaminergic neurons are located in the substantia nigra pars compacta. Over the last years various protocols for the production of DA neurons, for example, from embryonic stem cells or foetal neuronal stem cells, have been used. Another approach is the generation of DA neurons via induced pluripotent stem cells [3]. The use of conditionally immortalized progenitor cells is a promising approach due to nearly unlimited access of material [4]
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