gms | German Medical Science

Introduction: The purpose of the ongoing research is to improve our current skills and knowledge in stem cell isolation, cultivation and differentiation from the utricular and saccular epithelia of young mice.

Matherials and methods: We harvested utricles and sacculi from 7 days old NMRI mice. Utricles were trypsinized in order to isolate single cells. Obtained cells were cultivated at 37ºC and 5% CO2 in DMEM with F12 Nutrient mixture, B27, N2 supplement, IGF-1 and EGF. We mechanically dissociated primary spheres and cultivated them in the culture media mentioned above. Secondary spheres were placed on fibronectin coated tissue culture slide chambers in the absence of IGF-1 and EGF in order to observe their subsequent proliferation and differentiation. Cells were characterized by immunofluorescence for myosinVIIA (hair cell marker), nestin (intermediate filament VI marker) and beta III tubulin.

Results: We proved that vestibular epithelia contains pluripotent stem cells which were able to form cell clusters called spheres. Sphere-derived cells’ pluripotency was demonstrated by the expression of nanog, oct 4 and nestin markers (cell progenitors). Single spheres harvested and cultivated on fibronectin gave rise through differentiation to different cell types including neuron like-cells which were positive for myosin VIIA, nestin and beta III tubulin.

Conclusions: Utricular epithelia of seven days old mice contains sufficient pluripotent stem cells which, under special conditions, can generate spheres.

Cells obtained from utricular epithelia are pluripotent because they manifest markers fro nanog, oct 4 as well as markers for nestin gene, characteristic for cell progenitors.