Human rotation-mediated fetal mixed brain cell aggregate culture: characterization and N-methyl-D-aspartate toxicity.
One difficulty in generating in vitro models of neuropathogenesis lies in maintaining stable proportions of primary neurons within a mixed brain cell population. Rotation-mediated fetal brain aggregate culture has been modified to permit growth of human primary fetal brain cells containing 50 to 60% neurons. After 12 weeks cholinesterase, neuron specific enolase and microtubule-associated protein-2 were demonstrable by biochemical assay and immunocytochemical labelling of cryostat sections of human fetal brain aggregates. Upon exposure to the glutamate agonist; N-methyl-D-aspartate for 7 days at 35 days in vitro neuron specific enolase and cholinesterase decreased to 60 to 70% of untreated levels. Glial fibrillary acidic protein did not change significantly but swollen astrocytes were seen in labelled sections of treated aggregates. This method is useful to study human neurotoxicity and degeneration in mixed glial culture without astrocyte overgrowth.
Hayes
G M
GM
Department of Infectious Diseases, St George's Hospital Medical School, London, UK. ghayes@sghms.ac.uk
Fox
R M
RM
Cuzner
M L
ML
Griffin
G E
GE
D001253:Astrocytes / Q000166:cytology* / Q000187:drug effects; D001921:Brain / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D018690:Excitatory Amino Acid Agonists / Q000633:toxicity*; D005333:Fetus / Q000166:cytology; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D018613:Microscopy, Confocal; D016202:N-Methylaspartate / Q000633:toxicity*; D009410:Nerve Degeneration / Q000139:chemically induced; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D010751:Phosphopyruvate Hydratase / Q000032:analysis; D012399:Rotation
D018690:Excitatory Amino Acid Agonists; D016202:N-Methylaspartate; D007770:L-Lactate Dehydrogenase; D010751:Phosphopyruvate Hydratase