Investigation of neuronal precursor cells in different culture media and different periods of time

Introduction:

Perineural invasion is an adverse prognostic factor for the generation of head and neck cancers. The interaction of neuronal precursor cells (NPCs) and tumor cells (FaDus) can be studied in co-cultures with both cell lines. To elucidate the growth properties of neuronal precursor cells in vitro, we determined their growth pattern under various culture conditions.

Methods:

Neuronal precursor cells were obtained from GFP-transgenic mice. They were cultured as neurospheres for 24 or 96 hours in stem cell medium (Neurobasal, B27, retinoic acid, GFG2 und EGF) followed by culture in medium for tumor cells (RPMI-1640 medium containing 10% fetal calf serum) for further 24 or 96 hours. Finally, the cultured cells were fixed and stained for immunofluorescence with DAPI and antibodies raised against β-tubulin.

Results:

The results of the study show that the size of neurospheres and the length of their neurites depend on the length of cultivation. The longer the neuronal precursor cells were cultivated, the larger their size and length of neurites were detected. Neurospheres cultivated for 96 hours in stem cell medium followed by culture in RPMI-1640 medium for 24 hours showed a significant increase in the size of spheres. In addition, the neurites growing out of the spheres were longer and formed numerous cross-links. However, no further growth of neurites was observed upon cultivation of spheres for 8 days.

Conclusions:

Taken together, the study indicates that cultivating neuronal precursor cells increases their size and alters their shape, in particular upon subsequent culture in media designed for the cultivation of tumor cells.

Publication History

Publication Date:
18 April 2018 (online)