Differentiation of neuronal cells from NIH/3T3 fibroblasts under defined conditions
Differentiation of neuronal cells from NIH/3T3 fibroblasts under defined conditions
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DOI：
10.1111/j.1440-169x.2010.01235.x
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发表时间：
2011-04
期刊：
影响因子：
4.6
通讯作者：
Zhuo Wang;E. Sugano;H. Isago;T. Hiroi;M. Tamai;H. Tomita
中科院分区：
文献类型：
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作者：
Zhuo Wang;E. Sugano;H. Isago;T. Hiroi;M. Tamai;H. Tomita
我们试图测试分化的NIH/3 T3成纤维细胞是否可以在没有任何表观遗传修饰的情况下分化为神经元细胞。首先，进行了神经球测定，我们通过在神经干细胞培养基中漂浮NIH/3 T3成纤维细胞培养物成功地产生了神经球样细胞。这些球体在三次传代后能够形成亚球体，并表达神经祖细胞标志物Nestin、Sox 2、Pax 6和Musashi-1。第二，在转移到分化培养基并再培养8天后，这些球体中的细胞表达神经元标记物β-微管蛋白和神经丝200以及星形胶质细胞标记物胶质细胞酸性蛋白（GFAP）。最后，用全反式维甲酸和牛磺酸处理球后，β-微管蛋白的表达增加，并观察到光感受器标记物视紫红质和恢复素的染色。目前的研究表明，NIH/3 T3成纤维细胞可以在特定条件下产生神经球样、神经元样甚至感光细胞样细胞，这表明分化的非神经元细胞NIH/3 T3成纤维细胞，而不是多能细胞，如胚胎干细胞或诱导多能干细胞，可能具有转分化为神经元细胞的潜力，而无需添加任何表观遗传修饰剂。这种转分化可能是由于NIH/3 T3成纤维细胞在正常条件下保持休眠的可能的神经祖细胞潜能。
We attempted to test whether the differentiated NIH/3T3 fibroblasts could be differentiated into neuronal cells without any epigenetic modification. First, a neurosphere assay was carried out, and we successfully generated neurosphere‐like cells by floating cultures of NIH/3T3 fibroblasts in neural stem cell medium. These spheres have the ability to form sub‐spheres after three passages, and express the neural progenitor markers Nestin, Sox2, Pax6, and Musashi‐1. Second, after shifting to a differentiating medium and culturing for an additional 8 days, cells in these spheres expressed the neuronal markers β‐tubulin and neurofilament 200 and the astrocytic marker glial fibrillary acidic protein (GFAP). Finally, after treating the spheres with all‐trans retinoic acid and taurine, the expression of β‐tubulin was increased and the staining of photoreceptor markers rhodopsin and recoverin was observed. The present study shows that NIH/3T3 fibroblasts can generate neurosphere‐like, neuron‐like, and even photoreceptor‐like cells under defined conditions, suggesting that the differentiated non‐neuronal cells NIH/3T3 fibroblasts, but not pluripotent cells such as embryonic stem cells or induced pluripotent stem cells, may have the potential to be transdifferentiated into neuronal cells without adding any epigenetic modifier. This transdifferentiation may be due to the possible neural progenitor potential of NIH/3T3 fibroblasts that remains dormant under normal conditions.