The human NT-2 cell line as in vitro model system for the excitotoxic cascade during stroke [Elektronische Ressource] / von Francois Paquet-Durand

The human NT-2 cell line as in vitro model system for the excitotoxic cascade during stroke Vom Fachbereich Biologie der Universität Hannover zur Erlangung des Grades Doktor der Naturwissenschaften Dr. rer. nat. genehmigte Dissertation von Dipl. Biochem. François Paquet-Durand Geboren am 21. Februar 1968 in Kamen 2004 Referent: Prof. Dr. Gerd Bicker Koreferent: Priv. Doz. Dr. Bernhard Huchzermeyer Tag der Promotion: 7. 11. 2003 II Abstract Cells from the human teratocarcinoma line NTera-2 can be induced to terminally differentiate into postmitotic neurons when treated with retinoic acid. However, this differentiation process is rather time consuming as it takes between 42 and 54 days. In this study a modified differentiation protocol is introduced which reduces the time needed for differentiation considerably without compromising the quality of the neurons obtained. The introduction of a proliferation step as free floating cell spheres cuts the total time needed to obtain high yields of purified NT-2 neurons to about 24-28 days. The cells obtained show neuronal morphology and migrate to form ganglion-like cell conglomerates. Differentiated cells were characterised using immunocytochemical and histochemical techniques. Among others, the cells express neuronal polarity markers such as the cytoskeleton associated proteins MAP2 and Tau.