Mouse Embryonic Stem Cells Differentiation to Neuron-like Cells

Unique features of embryonic stem cells (ESCs) such as unlimited proliferation and differentiation into other types of cells make them a favorable tool for biomedical researches as well as a potential source for therapeutic application for neurodegenerative diseases, such as Parkinson’s and Alzheimer’s diseases. In recent years, in vitro methods have been developed which permit the growth of neurons from pluripotent cells in culture. These cells can be maintained as stable, proliferative and undifferentiated cell lines if cultured on feeder layer or in presence of leukemia inhibitory factor. Since ESCs can be proliferated and differentiated, it is possible to generate large numbers of donor cells for neural transplantation. Retinoic acid (RA) is one of the most important morphogenesis, and its embryonic distribution correlates with neural differentiation and positional specification in the developing central nervous system.
After proliferation on the mouse embryonic fibroblast feeder cells in the presence of LIF, for the study of CCE cell line differentiation these cells were cultured to producing cell aggregates (embryoid bodies). The embryoid bodies were under the protocol 4- / 4 (four days in the presence or absence of retinoic acid) at concentration of 10-6 µM retinoic acid for differentiation. Then morphological, molecular and immunocytochemistry examination were used to assess neurological factors.
In this induction protocol, highly proportion (%80) of ESCs could be induced to differentiation into neuron-like cells. The cells expressed neuroepitelial cell marker nestin. In addition, the results indicated that RA could induce nerve growth factor gene expression in the ESCs.
These findings suggest that RA strictly regulates the neuralization and specification during differentiation of mouse ESCs, especially for the differentiation into nerve cells.