Production of stable GFP-expressing neural cells from P19 embryonal carcinoma stem cells

- Using P19 EC cell line, stable GFP-expressing cells was made and characterized.
- Neural differentiation of GFP+ cells was induced by selegiline, a drug used in the treatment of Parkinson's.
- These morphologically neural cells were immunoreactive to neuron specific proteins.
- The differentiated cells expressed neuroepithelial and neural specific markers.
- GFP+ cells were able to survive, migrate and differentiate in chick embryo CNS.

Murine P19 embryonal carcinoma (EC) cells are convenient to differentiate into all germ layer derivatives. One of the advantages of P19 cells is that the exogenous DNA can be easily inserted into them. Here, at the first part of this study, we generated stable GFP-expressing P19 cells (P19-GFP+). FACS and western-blot analysis confirmed stable expression of GFP in the cells. We previously demonstrated the efficient induction of neuronal differentiation from mouse ES and EC cells by application of a neuroprotective drug, selegiline In the second part of this study selegiline was used to induce differentiation of P19-GFP+ into stable GFP-expressing neuron-like cells. Cresyl violet staining confirmed neuronal morphology of the differentiated cells. Furthermore, real-time PCR and immunoflourescence approved the expression of neuron specific markers. P19-GFP+ cells were able to survive, migrate and integrated into host tissues when transplanted to developing chick embryo CNS. The obtained live GFP-expressing cells can be used as an abundant source of developmentally pluripotent material for transplantation studies, investigating the cellular and molecular aspects of early differentiation.

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