Differentiation of green fluorescent protein-labeled embryonic stem cell-derived neural precursor cells into Thy-1-positive neurons and glia after transplantation into adult rat striatum
Differentiation of green fluorescent protein-labeled embryonic stem cell-derived neural precursor cells into Thy-1-positive neurons and glia after transplantation into adult rat striatum
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DOI：
10.3171/jns.2000.93.6.1026
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发表时间：
2000-12-01
影响因子：
4.1
通讯作者：
Addicks, K
中科院分区：
文献类型：
--
作者：
Arnhold, S;Lenartz, D;Addicks, K
Object.本研究的目的是评估有关移植胚胎干细胞（ESC）衍生的神经元细胞迁移到宿主脑中的能力的新信息，并评估这些细胞作为神经退行性疾病如帕金森病（PD）的细胞替代疗法的可能来源。作者通过使用D3来源的ESC克隆研究了ESC来源的神经前体细胞在宿主纹状体中迁移和分化的能力，所述D3来源的ESC克隆用鸡β-肌动蛋白巨细胞病毒增强子驱动的绿色荧光蛋白（GFP）标记的构建体稳定转染。由于供体细胞的绿色荧光标记，该程序允许容易地监测所有移植的细胞。这种方法也提供了简单的估计细胞整合和同时观察整个移植的细胞群体的免疫细胞化学鉴定的神经元和神经胶质细胞分化。在合成培养基中选择巢蛋白阳性神经前体细胞后，将其植入雄性成年Wistar大鼠的纹状体中。在移植后3天至4周进行形态学研究，分析其整合情况。研究人员发现，移植后，GFP标记的细胞亚群分化成各种神经形态类型，这些类型对小鼠特异性Thy-1抗原呈阳性，已知Thy-1抗原在神经元上表达，并且对星形胶质细胞标记物胶质细胞酸性蛋白呈阳性。此外，对这些标记物中的任何一个呈阴性的GFP表达细胞保持接近注射部位，推测代表神经谱系的其他衍生物。总之，这些研究结果有助于基础研究有关未来的移植策略在神经退行性疾病，如PD。
Object. The aim of this investigation was to assess new information concerning the capacity of transplanted embryonic stem cell (ESC)-derived neuronal cells to migrate into host brain and to evaluate these cells as a possible source for cell replacement therapy in neurodegenerative disorders such as Parkinson's disease (PD).Methods. The authors investigated the ability of ESC-derived neural precursor cells to migrate and differentiate in a host striatum by using a D3-derived ESC clone that was transfected stably with a chicken beta -actin cytomegalovirus enhancer-driven green fluorescent protein (GFP)-labeled construct. This procedure allowed easy monitoring of all transplanted cells because of the green fluorescent labeling of donor cells. This approach also afforded easy estimation of cell integration and simultaneous observation of the entire transplanted cell population in relation to immunocytochemically identified neuronal and glial differentiation. After selection of nestin-positive neural precursor cells in a synthetic medium, they were implanted into the striatum of male adult Wistar rats. Their integration was analyzed on morphological studies performed 3 days to 4 weeks posttransplantation.Conclusions. The investigators found that after transplantation, a subpopulation of GFP-labeled cells differentiated into various neural morphological types that were positive for the mouse-specific Thy-1 antigen, which is known be expressed on neurons, as well as being positive for the astroglial marker glial fibrillary acidic protein. Moreover, GFP-expressing cells that were negative for either of these markers remained close to the injection site, presumably representing other derivatives of the neural lineage. Together, these findings contribute to basic research regarding future transplantation strategies in neurodegenerative diseases such as PD.