Kan Inna, Ben-Zur Tali, Barhum Yael, Levy Yossef S, Burstein Alex, Charlow Tirza, Bulvik Shlomo, Melamed Eldad, Offen Daniel
Laboratory of Neurosciences, Felsenstein Medical Research Center, Rabin Medical Center, Beilinson Campus, Tel Aviv University, Petah Tiqwa 49100, Israel.
Neurosci Lett. 2007 May 23;419(1):28-33. doi: 10.1016/j.neulet.2007.03.070. Epub 2007 Apr 13.
Parkinson's disease (PD) is a neurodegenerative disorder, caused by a selective loss of dopaminergic neurons in the substantia nigra. In PD, the best therapeutic modalities cannot halt the degeneration. The selective hallmark pathology and the lack of effective treatment make PD an appropriate candidate for cell replacement therapy. Adult autologous bone-marrow-derived mesenchymal stem cells (MSCs) have been investigated as candidates for cell replacement strategies. Several laboratories, including ours, have induced MSCs into neuron-like cells demonstrating a variety of neuronal markers including dopaminergic characteristics, such as the expression of tyrosine hydroxylase (TH). This project aimed to induce MSCs into mature dopamine secreting cells and to generate a bioassay to evaluate the induction. For that purpose, we created a reporter vector containing a promoter of TH, the rate-limiting enzyme in the dopamine synthesis and red fluorescent protein DsRed2. Transfection of human neuroblastoma, dopamine synthesizing, SH-SY5Y cells confirmed the reliability of the constructed reporter plasmid. Following dopaminergic differentiation of the transfected human MSCs cells, TH expressing cells were identified and quantified using flow cytometry. Further study revealed that not only did the differentiated cells activate TH promoter but they also expressed TH protein and secreted dopamine. The reported results indicate that MSCs may be primed in vitro towards a dopaminergic fate offering the promise of innovative therapy for currently incurable human disorders, including PD.
帕金森病（PD）是一种神经退行性疾病，由黑质中多巴胺能神经元的选择性丧失引起。在帕金森病中，最佳治疗方式也无法阻止神经退变。这种选择性标志性病理特征以及缺乏有效治疗方法，使得帕金森病成为细胞替代疗法的合适候选对象。成人自体骨髓间充质干细胞（MSCs）已被作为细胞替代策略的候选细胞进行研究。包括我们实验室在内的几个实验室，已将间充质干细胞诱导为神经元样细胞，这些细胞表现出多种神经元标志物，包括多巴胺能特性，如酪氨酸羟化酶（TH）的表达。本项目旨在将间充质干细胞诱导为成熟的多巴胺分泌细胞，并建立一种生物测定法来评估诱导效果。为此，我们构建了一个报告载体，其包含多巴胺合成限速酶TH的启动子以及红色荧光蛋白DsRed2。转染人神经母细胞瘤、多巴胺合成细胞系SH-SY5Y细胞，证实了所构建报告质粒的可靠性。在转染后的人间充质干细胞进行多巴胺能分化后，使用流式细胞术对表达TH的细胞进行鉴定和定量。进一步研究表明，分化后的细胞不仅激活了TH启动子，还表达TH蛋白并分泌多巴胺。报告结果表明，间充质干细胞在体外可能被诱导为多巴胺能命运，这为包括帕金森病在内的目前无法治愈的人类疾病提供了创新治疗的希望。
