Primary dissociated cultures of human brainstem cells: a useful tool for their characterization and neuroprotection study

Dissociated cell cultures were prepared from brainstems of 5- to 10-week-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 5HT-stained cells were characterized in the rhombencephalic cultures, representing respectively the fetal localization of the locus coeruleus and raphe nuclei. DA-stained cells were characterized in the mesencephalic cultures; these DA-cells originating from the substantia nigra presented morphological aspects different from the DA-rhombencephalic cells. Two types of GABA neurons and glial cells presenting glial fibrillary acidic protein (GFA-P) reactivity were also found in all the cultures. Two non-competitiveN-methyl-D-aspartate antagonists, 1-[1-(2-thienyl)cyclohexyl]piperidine (TCP) andcis-Pip/Me 1-[1-(2-thienyl)-2-methylcyclohexyl]piperidine (GK11) in enantiomeric form (–), have been investigated 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. Thisin 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.Abbreviations CA catecholamine - DA dopamine - CNS central nervous system - GFA-P glial fibrillary acidic protein - IR immunoreactivity - NMDA N-methyl-d-aspartate - NA noradrenaline - 5HT serotonin - TCP 1-[1-(2-thienyl)cyclohexyl]piperidine - GK11 cis-Pip/Me 1-[1-(2-thienyl)-2-methylcyclohexyl]piperidine