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

• Published in: Cell biology and toxicology 1995-08, Vol.11 (3-4), p.155-160
• Main Authors: LEVALLOIS, C, CALVET, M. C, KAMENKA, J. M, PETITE, D, PRIVAT, A
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Brain Stem - cytology; Brain Stem - drug effects; Brain Stem - growth & development; Cell Culture Techniques; Cells, Cultured; Fetus; General pharmacology; Humans; Immunohistochemistry; Medical sciences; Neurons - cytology; Neurons - drug effects; Pharmacokinetics. Pharmacogenetics. Drug-receptor interactions; Pharmacology. Drug treatments; Rhombencephalon - cytology; Rhombencephalon - drug effects; Rhombencephalon - growth & development
• ISSN: 0742-2091; 1573-6822
• DOI: 10.1007/BF00756517

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-competitive N-methyl-D-aspartate antagonists, 1-[1-(2-thienyl)cyclohexyl]piperidine (TCP) and cis-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. 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.