Human Embryo Neuronal Culture <i>in Vitro</i>: A Model to Study Cellular Physiology, Receptors, Power and Toxicity of Cytostatic Drugs for Human Use

Abstract

Neural cells cultures from human embryo brain of 9˚ -11˚W gestational age have been used to study ERα (Estrogens Receptor α) and to perform toxicity test for Mitomycin C and Methotrexate. Histochemical confirmation of cellular neuronal phenotype was based on histochemical evidence of NSE (Neuron Specific Enolase).The detection of ERα in neuronal cells was performed with a rabbit Monoclonal Antibody. ERα was absent both on neurons grown in vitro and on tissue brain specimens. This finding is apparently in contrast with the positive immunoreactivity of ERα and ERβ reported by other Authors on foetal and adult CNS (Central Nervous System). The absence of nuclear ERα on neurons in culture and in brain tissue specimens in our experiment is not in contrast with the relevant physiologic role of estrogens on nervous central system, but it could be correlated to the embryonic period of life and could represent a protection of male brain from an undue estrogens imprinting. The mitomycin C, alkylation agent, has shown in our experiment a major neurotoxic and cytostatic power in comparison with methotrexate. Our conclusion is that human embryo neuronal culture in vitro is a powerful instrument for physiology and human therapy for cancer and neurodegenerative diseases.

Growing Human Embryonic Neurons in Glass Capillary: A Simple Method to Study Interaction between Motile Cells and Neurons and to Explore the Neurotoxicity of Metals

“…A culture of embryonic neurons derived from human embryos of 7 -10 weeks was prepared according to the procedures used in our laboratory [5]. A confluent neuronal monolayer was treated with mild trypsinization (1 cc of 0.05% trypsin-EDTA Gibco for 1 min); after washing, neurons detached from the monolayer were suspended in 0.5 cc of Chang Medium (Irvine Scientific), were aspirated into a 1 cc syringe and injected into a clean, sterilized glass microhematocrit capillary tube.…”