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The population of stem/progenitor cells was selected from human cord blood non-hematopo- ietic (CD34 and CD45 negative) mononuclear fraction and was shown to attain neural features. Due to repeated expansion and selection of these cells in the presence of EGF we have established the first clonogenic, non-immortalized human umbilical cord blood neural stem cell like line (HUCB-NSC). This line can be maintained in culture at different developmental stages and their fate decisions can be experi- mentally manipulated in vitro by the presence of trophic factors, mitogenes and neuromorphogenes. Standardized conditions for their growth and differentiation have been established. The activation of the WNT, DELTA/NOTCH and FGFR1 signaling pathways in HUCB-NSC was shown by molecular and immunocytochemical analysis. Differentiation in the presence of dBcAMP directed HUCB-NSC predo- minantly (80%) into neuronal lineage, as revealed by DNA microarray and immunocytochemistry. This included expression of several functional proteins: glutamatergic, GABA-ergic, dopamine, serotonin and acetylcholine receptors or synaptic vesicle proteins. That was further confirmed by electrophysiological studies showing in differentiated HUCB-NSC two types of voltage-sensitive and several ligand gated currents typical for neuronal cells. Obtained results confirm the stem character and neural commitment of HUCB-NSC cell line. This is the first report of establishment and functional neuronal differentiation of nonimmortalized somatic stem cell line.
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Andrzej Łukaszyk - przewodniczący, Zofia Bielańska-Osuchowska, Szczepan Biliński, Mieczysław Chorąży, Aleksander Koj, Włodzimierz Korochoda, Leszek Kuźnicki, Aleksandra Stojałowska, Lech Wojtczak

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