Morphological and morphometric analysis of serotonin-containing neurons in primary dissociated cultures of human rhombencephalon: a study of development.

Primary dissociated cultures of rhombencephalon were prepared from 5-9-week-old human fetuses. Half of some cultures were treated by two non-competitive N-methyl-D-aspartate antagonists, namely 1-(2-thienyl)cyclohexylpiperidine (TCP) and cis-Pip/Mel-[1-(2-thienyl)-2-methyl-cyclohexyl]piperidine (GK11) in negative enantiomeric form, which enhance the survival of human fetal central nervous system cells in culture. At different days in vitro, the treated and the control cultures were processed for immunocytochemical detection of serotonin-containing neurons which were studied by morphological and morphometric analysis. Statistical analysis showed that the surface of the stained neurons increased as a function of two parameters of time, the gestational age of the cells and the duration of the cultures. The complexity of the shape of the serotonin neurons characterized by the shape factor, the number of bifurcations and the morphological feature (bipolar or multipolar) was found to increase with the gestational age. It appears that the in vitro development of the embryonic cells which represents stages of maturation and differentiation can be specifically evaluated. Such an analysis of fetal central nervous system cells improves the knowledge of factors important in grafting experiments. We verified that the two drugs do not appreciably alter the in vitro development of the treated cells; thus they may be considered as promising drugs for human neuroprotection.