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Differentiation of neuronal cells from NIH/3T3 fibroblasts under defined conditions
Author(s) -
Wang Zhuo,
Sugano Eriko,
Isago Hitomi,
Hiroi Teru,
Tamai Makoto,
Tomita Hiroshi
Publication year - 2011
Publication title -
development, growth and differentiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.864
H-Index - 66
eISSN - 1440-169X
pISSN - 0012-1592
DOI - 10.1111/j.1440-169x.2010.01235.x
Subject(s) - neurosphere , nestin , transdifferentiation , microbiology and biotechnology , biology , sox2 , neural stem cell , induced pluripotent stem cell , progenitor cell , stem cell , embryonic stem cell , p19 cell , adult stem cell , biochemistry , gene
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.