gms | German Medical Science

83. Jahresversammlung der Deutschen Gesellschaft für Hals-Nasen-Ohren-Heilkunde, Kopf- und Hals-Chirurgie e. V.

Deutsche Gesellschaft für Hals-Nasen-Ohren-Heilkunde, Kopf- und Hals-Chirurgie e. V.

16.05. - 20.05.2012, Mainz

Vestibular stem cells

Meeting Abstract

  • corresponding author Roxana Vintila - ENT Bega, Timisoara, Rumanien
  • Oana Gavriliuc - University of Medicine and Pharmacy "Victor Babes", Timisoara, Romania
  • Florina Bojin - University of Medicine and Pharmacy "Victor Babes" Timisoara, Timisoara, Romania
  • Elen Gocza - MBK Godollo, Godollo, Hungary
  • Gheorghe Iovanescu - ENT clinic Bega, University of Medicine and Pharmacy "Victor Babes"Timisoara, Timisoara, Romania
  • Calin Tatu - University of Medicine and Pharmacy "Victor Babes"Timisoara, Timisoara, Romania
  • Cornelia Vintila - Agricultural Sciences University Timisoara, Timisoara, Romania
  • Stan Cotulbea - University of Medicine and Pharmacy "Victor Babes" Timisoara, ENT Clinic Bega, Timisoara, Romania

Deutsche Gesellschaft für Hals-Nasen-Ohren-Heilkunde, Kopf- und Hals-Chirurgie. 83. Jahresversammlung der Deutschen Gesellschaft für Hals-Nasen-Ohren-Heilkunde, Kopf- und Hals-Chirurgie. Mainz, 16.-20.05.2012. Düsseldorf: German Medical Science GMS Publishing House; 2012. Doc12hnod753

doi: 10.3205/12hnod753, urn:nbn:de:0183-12hnod7539

Veröffentlicht: 4. April 2012

© 2012 Vintila et al.
Dieser Artikel ist ein Open Access-Artikel und steht unter den Creative Commons Lizenzbedingungen (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.de). Er darf vervielf&aauml;ltigt, verbreitet und &oauml;ffentlich zug&aauml;nglich gemacht werden, vorausgesetzt dass Autor und Quelle genannt werden.


Gliederung

Text

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 sacculus 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. Sphere pluripotency was established with the help of stem cell markers Nanog and Oct-4. We mechanically dissociated primary spheres and cultivated them in the culture media mentioned above. Secondary spheres were placed on fibronectin coated nuncs in the absence of IGF-1 and EGF in order to observe their differentiation. For cell characterization we performed immunoflorescence and immunohistochemistry for myosinVIIA (hair cell marker), nestin (intermediate filament VI marker), beta III tubulin (neuronal marker).

Results: We proved that utricular and saccular epithelia contain pluripotent stem cells which were able to form spheres. Sphere cells pluripotency was proved with help of nanog, oct 4, nestin marker expression, characteristic for cell progenitors.

Also sphere dissociation and separate cell cultivation lead to formation of a higher amount of spheres, proving that sphere cells are pluripotent and capable of renewal. Single spheres cultivated on fibronectin differentiated to different cell types including neuron like- cells positive for beta III tubulin, cells positive for myosin VIIA and nestin.

Conclusions: Utricular epithelia of seven days old mice contains sufficient pluripotent stem cells which generate spheres and their differentiation generate nestin, betaIII tubulin and myosin VIIA expressing cells.