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 m aintaining stable proportions of primary neurons within a mixed brain cell population. Rotation-mediated fetal brain aggregate culture has been modifi ed to permit growth of human primary fetal brain cells containing 50 to 60% neurons. After 12 weeks cholinesterase, neuron specific enolase and microt ubule-associated protein-2 were demonstrable by biochemical assay and immun ocytochemical labelling of cryostat sections of human fetal brain aggregate s. 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 cha nge significantly but swollen astrocytes were seen in labelled sections of treated aggregates. Th is method is useful to study hu man neurotoxicity an d degeneration in mixed glial culture without astrocyte overgrowth. (C) 200 0 Elsevier Science Ireland ltd. All rights reserved.