PRIMARY DISSOCIATED CULTURES OF HUMAN BRAIN-STEM CELLS - A USEFUL TOOL FOR THEIR CHARACTERIZATION AND NEUROPROTECTION STUDY

Dissociated cell cultures were prepared from brainstems of 5- to 10-we ek-old human fetuses. Catecholamine- as well as indolamine-containing cells were visualized using respectively dopamine (DA), noradrenaline (NA) and serotonin (5HT) as immunocytochemical markers. NA-,DA-,and 5H T-stained cells were characterized in the rhombencephalic cultures, re presenting respectively the fetal localization of the locus coeruleus and raphe nuclei. DA-stained cells were characterized in the mesenceph alic cultures; these DA-cells originating from the substantia nigra pr esented morphological aspects different from the DA-rhombencephalic ce lls. Two types of GABA neurons and glial cells presenting glial fibril lary acidic protein (GFA-P) reactivity were also found in all the cult ures. Two non-competitive N-methyl-D-aspartate antagonists, 1-[1-(2-th ienyl)cyclohexyl]piperidine (TCP) and cis-Pip/Me 1-[1-(2-thienyl)-2-me thylcyclohexyl]piperidine (GK11) in enantiomeric form (-), have been i nvestigated for survival on rhombencephalic cultured cells. The number of 5HT-cells was found to be greater in the treated cultures than in the control ones. This in vitro system appears to be a useful tool for the investigation of the development of central nervous system (CNS) cells as well as the study of neuroprotection.